Pharmaceutical Biology
2004, Vol. 42, Supplement, pp. 46–63
Natural Product Polyphenols of Relevance to Human Health
Tamara P. Kondratyuk and John M. Pezzuto
Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy and Pharmaceutical Science,
Purdue University, West Lafayette, Indiana, USA
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Abstract
Polyphenols are widely distributed in the plant kingdom
and represent an abundant antioxidant component of the
human diet. The review offers a brief description of the
chemistry and occurrence in plant food of four important
groups of polyphenols: phenolic acids, flavonoids, stil-
benes, and lignans. Interest in the possible health benefits
of polyphenols has increased due to corresponding anti-
oxidant capacity. Considerable evidence is now available
showing anticarcinogenic effects of polyphenolic com-
For personal use only.
pounds, as well as potential to prevent cardiovascular
and cerebrovascular diseases. The use of botanicals has
received a large amount of attention in recent years.
Epidemiological studies have shown that a reduced risk
of cancer is associated with diets rich in vegetables and
fruits, and methods for the discovery and characteri-
zation of active compounds from plant sources are
available. These results are promising, but additional
research on the molecular mechanism of action of poly-
phenols and their application to human health is
required.
Keywords: Antioxidants, chemoprevention, drug dis-
covery, flavonoids, medicinal botanicals, phenolic acids,
phytochemicals, polyphenols.
Introduction
Polyphenols form a complex group of molecules associa-
ted with most plant cell walls. They range in chemical
complexity from simple phenolic acids (e.g., caffeic acid)
to high-molecular-weight tannins. More than 8000 phe-
nolic and polyphenolic compounds have been identified
in various plant species, where they have several impor-
tant functions. Examples include inhibition of pathogen
Accepted: September 15, 2004
Address correspondence to: John M. Pezzuto, Purdue University, College of Pharmacy, Nursing and Health Sciences, Department of
Medicinal Chemistry and Molecular Pharmacology, Room 104, Heine Pharmacy Building, 575 Stadium Mall Drive, West Lafayette,
IN 47907-2051, USA. Tel: (765) 494-1368; Fax: (765) 494-7880; E-mail: jpezzuto@purdue.edu
DOI: 10.1080/13880200490893519 # 2004 Taylor & Francis Ltd.
development, decay of microorganisms, and protection
against UV radiation, photosynthetic stress, reactive
oxygen species, wounds, and herbivores. Some polyphe-
nols are essential for plant physiology, being involved
in diverse functions such as pigmentation, pollination,
enhanced predator resistance (by acting as phytoalexins),
or increased astringency, thus making plants unpalatable
as foods. They protect crops from plague and preharvest
seed germination. Polyphenols have many industrial
applications, such as in the production of paints, paper,
cosmetics, as tanning agents, and in the food industry as
natural colorants and preservatives. In addition, some
phenolic compounds are antibiotics and antidiarrheal,
antiulcer, and anti-inflammatory agents. They can be
used in the treatment of diseases such as hypertension,
vascular fragility, allergies, and hypercholesterolemia
(Bravo, 1998; Higdon & Frei, 2003).
Chemoprevention is a promising new approach in
cancer research wherein agents are administered to
inhibit, delay, or reverse the process of carcinogenesis.
Epidemiological data have suggested that high consump-
tion of vegetables and fruits is associated with a low risk
of cancer and cardiovascular diseases (American Insti-
tute of Cancer Research=World Cancer Research Fund,
1997; van Duyn & Pivonka, 2000; Byers, 2002; Bub et al.,
2003), whereas a high intake of red meat may increase
some types of cancer (Hollman et al., 1999). A variety
of in vitro studies have shown that polyphenols medi-
ate antioxidative and immunomodulatory activities
(Middleton, 1998; Ross & Kasum, 2002; Dragsted,
2003). Further, polyphenols have potential to prevent
genotoxicity by reducing exposure to oxidative and carci-
nogenic factors (Pool-Zobel et al., 1999). There has been
considerable scientific interest in the possibility that
increased intake of dietary antioxidants may protect
 Natural product polyphenols of relevance to human health
against some chronic diseases. Among the dietary phe-
nolic compounds, flavonoids have extensively been
studied, and there are several reviews relating to the anti-
oxidant activity of these compounds (Lairon & Amiot,
1999; Chan et al., 2003; Steinberg et al., 2003; Urso &
Clarksonn, 2003).
There is growing interest in medicinal botanicals as
part of complementary medicine in the United States.
Approximately 40% of Americans use alternative rem-
edies, including herbal medicine, for disease prevention
and therapy (Eisenberg et al., 1998). The high cost, side
effects, and therapeutic limitations of conventional med-
ications are key factors that drive the revival of herbal
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remedies. People can easily obtain herbal medicines or
botanical supplements on an over-the-counter basis and
feel comfortable consuming these products, even though
safety and efficacy are generally not established on a rig-
orous scientific basis. This may be problematic due to
factors such as drug-drug interactions (von Gyuenigen
& Hopkins, 2000; Go et al., 2001). On the other hand,
these preparations may mediate beneficial responses,
either due to active chemical constituents or placebo
effects. In any case, botanicals have been used by human
beings throughout history.
For personal use only.
Conventional medical literature also documents the
widespread use of herbal remedies in many countries
throughout the world, and the presence of certain phy-
tochemical components supports the pharmacological
and physiological efficacy of some ethnomedical treat-
ment regimens. The cancer inhibitory potential of
human nutrients derived from plants, as well as of
non-nutrient constituents of phytochemicals, has been
confirmed in various animal models (Pezzuto, 1997).
For more than 30 years, the prevention of cancer has
been emphasized as a preferable option relative to
chemotherapy. Significant success has been realized
through the control of tobacco-related cancers by means
of education and prohibition of smoking in various pub-
lic settings. In addition, cancer chemoprevention has
been developed as a major field of scientific investi-
gation. Accordingly, a great deal of evidence has sug-
gested the protective role of a diet rich in fruits and
vegetables (van Duyn & Pivonka, 2000; Terry et al.,
2001). Overall, diets high in vegetables and fruits (more
than 400 g=day) may prevent at least 20% of all cancers.
Some of the most convincing evidence for the health
benefits of fruit and vegetable consumption relates to
the reduced risk of gastrointestinal cancers, such as
those associated with the mouth, pharynx, esophagus,
stomach, colon, and rectum. Vegetables are more effec-
tive than fruits (Terry et al., 2001). The mechanisms by
which vegetables and fruits reduce cancer are likely to
be multiple and complex. Various stages of carcinogen-
esis may be inhibited, and various in vitro or in vivo sys-
tems may be used to model these inhibitory effects. It is
logical to acquire compelling in vitro data prior to
47
performing tests with animal models, and it is generally
necessary to isolate and characterize active chemical
principles.
The current review is focused on characterization,
bioavailability, and cancer chemopreventive potential
of several classes of polyphenols. In addition, we describe
the process of cancer chemopreventive drug discovery
from plant sources with characterization of experimental
approaches, with special emphasis on work performed in
our laboratory.
Nature and occurrence of phenolic
phytochemicals
Phenolic compounds comprise one of the largest groups
of plant metabolites, and they are an important part of
human diets. All plant phenolic compounds arise from
a common intermediate, phenylalanine, or a close pre-
cursor, shikimic acid. They arise biogenetically from
two main synthetic pathways: the shikimate pathway
and the acetate pathway (Herrmann, 1995). Plants poly-
phenols can range from simple species, such as phenolic
acids, to highly polymerized compounds, such as tan-
nins. They occur primarily in conjugated forms, with
one or more sugar residues linked to hydroxyl groups,
although direct linkages of the sugar to an aromatic
carbon also exists. Sugars can be monosaccharides, disac-
charides, or oligosaccharides. Glucose is the most
common sugar residue. Association with other com-
pounds, such as carboxylic and organic acids, amines,
and lipids, and linkages with other phenols are also com-
mon. They can be divided into at least 10 different classes
based on their general chemical structure (Bravo, 1998).
For this review, we have chosen four groups for dis-
cussion, with emphasis on their possible applications
for cancer prevention. Flavonoids and phenolic acids
most commonly are widespread in foods.
Phenolic acids
Hydroxybenzoic acids and hydroxycinnamic acids
account for about a third of the phenolic compounds
in our diet. The most frequently encountered species is
caffeic acid (Fig. 1). Caffeic acid is found in many diet-
ary substances, such as apples, plums, tomatoes, and
grapes. Derivatives of hydroxycinnamic acid are found
in nearly every plant. One of the most widespread repre-
sentatives is curcumin (Fig. 1). It is widely used as a
food preservative and yellow coloring agent for foods,
drugs, and cosmetics. Phenolic acids can act as anti-
oxidants by a number of pathways, but perhaps the
most significant is by free-radical scavenging in which
the phenolic compound can quench the free-radical
reaction.
 48 T.P. Kondratyuk and J.M. Pezzuto
Figure 1. Phenolic acids and derivatives.
Flavonoids
The flavonoids are a complex group of polyphenolic
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plant metabolites found in the food of human beings
(Havsteen, 2002). All share a three-ring structure of
two aromatic centers, rings A and B, and a central oxy-
genated heterocycle moiety, ring C (Fig. 2). Biogeneti-
cally, the A ring usually arises from a molecule of
resorcinol, and the B ring is derived from the shikimate
pathway. Flavonoids can further be subdivided into six
major subclasses, based on variations in the heterocyclic
C-ring, including flavones, flavonols, flavonones, cate-
chins, anthocyanidins, and isoflavones (Ross & Kasum,
2002). Flavonoids are widely distributed in beverages
For personal use only.
of plant origin, such as tea, cocoa, and wine. Tea and
onions are the main dietary sources of flavonols and fla-
vones. It has been estimated that, even in industrialized
societies, intakes of flavonoids could be as high as 1 g
per day, but this is probably an overestimate based on
inadequate analytical data (Hertog & Hollman, 1996).
A substantial body of experimental work has established
that flavonoids can suppress carcinogenesis in animal
models, and there is considerable interest in the biologi-
cal effects of these compounds at the cellular level.
Flavonoids interact with cellular signal pathways that
control the cell cycle, differentiation, and apoptosis.
Their antineoplastic effects can involve antioxidant
activity, induction of phase II enzyme activity, inhibition
of protein kinases, and interaction with type II estrogen
binding sites (Gee & Johnson, 2001).
Stilbenes
Stilbenes contain two phenyl moieties connected by a
two-carbon methylene bridge (Fig. 3). Most stilbenes in
plants act as antifungal phytoalexins, compounds that
Figure 2. Basic flavonoid structure.
are synthesized only in response to infection or injury.
The most extensively studied stilbene is resveratrol.
Resveratrol was first recognized as a biologically active
compound by Siemann and Creasy (1992). Resveratrol
(trans-3,5,40 -trihydroxystilbene) is a phytoalexin, or a
class of antibiotics of plant origin, produced primarily in
grapes and peanuts as edible plants. The highest concen-
tration of resveratrol (50–100 mg=g) is found in the skin
of the grape. The reason for synthesis of resveratrol in
grapes is associated with natural stress factors such as gray
mold or fungal infections. It is considered to be a part of
the plant defense system (Schwekendiek et al., 1992).
Significant amounts of resveratrol were detected in
healthy fruit clusters, prior to visual detection of mold
lesions, thus suggesting resveratrol was synthesized soon
after recognition of the pathogen by the plant (Jeandet
et al., 2002). It is conceivable that resveratrol mediates
beneficial effects with human cells by mechanisms similar
to those mediated in plant cells.
Resveratrol has been shown to have anticancer (che-
mopreventive), anti-inflammatory, antifungal, and anti-
microbial properties. It has a long history of being used
as an herbal remedy, especially in oriental medicine for
the treatment of lipid, inflammatory, and heart disor-
ders. Most early studies were associated with antioxidant
or cardiovascular activities and ‘‘stickiness’’ of blood
platelets. Resveratrol has been reported to have a diverse
range of pharmacological properties, including anti-
inflammatory, estrogen receptor agonist (Gehm et al.,
1997), and effects on cell signaling pathways, cell pro-
liferation, tumor growth, and apoptosis (Stewart et al.,
1999). Some experiments suggest that resveratrol inhi-
bits the development of cancer and can be used as a
chemopreventive agent, but metabolic pathways and
the biological effects of metabolites remain to be charac-
terized (Bhat et al., 2001; Bhat & Pezzuto, 2002).
Lignans
Lignans are diphenolic compounds that contain a 2,3-
dibenzylbutane structure that is formed by the dimeriza-
tion of two cinnamic acid residues (Fig. 3). Lignans are
naturally occurring chemicals that are widespread within
plants. Several lignans, such as secoisolariciresinol, are
 Natural product polyphenols of relevance to human health
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Figure 3. Representative structures of stilbenes and lignans.
considered to be phytoestrogens. These are especially
abundant in flax seed. Bacteria in human intestines con-
vert them into two other lignans, enterolactone and
enterodiol, which also have estrogen-like effects. Lignans
are being studied for possible use in cancer prevention,
particularly against breast cancer. If there is little estro-
gen in the body, lignans may act like a weak estrogen,
but when natural estrogen is abundant in the body,
For personal use only.
lignans may reduce the effect of estrogen by displacing
it from cells. This displacement of the hormone may help
prevent some cancers, such as breast cancer, that depend
on estrogen. Very early evidence suggests that lignans
may also be antioxidants, although the strength of their
antioxidant activity is not clear (Kitts et al., 1999).
Polyphenols as antioxidants
Plant polyphenols are well recognized for their antioxi-
dant activities. These compounds scavenge free radicals
and disrupt the free-radical chain reaction of lipid per-
oxidation. These antioxidants can be classified as
water-soluble or lipid-soluble, depending on whether
they act primarily in the aqueous phase or in the lipophi-
lic region of cell membranes. Hydrophilic antioxidants
include ascorbic acid and urate. Ubiquinols, retinoids,
carotenoids, flavonoids, and tocopherol are representa-
tive lipid-soluble antioxidants. Plasma proteins, glu-
tathione, and urate are endogenous, whereas ascorbic
acid, carotenoids, retinoids, flavonoids and tocopherols
constitute some of the dietary antioxidants. Certain rad-
ical scavengers are not recyclable, whereas others are
recycled through the intervention of a series of enzyme
systems or other nonenzymic antioxidant systems. Diet-
ary exposure to flavonoids is significant. As noted above,
the average diet in the U.K. and the U.S. may contain up
to 1 g of mixed flavonoids per day. Dietary intake far
exceeds that of vitamin E, a monophenolic antioxidant,
and of b-carotene (Kandaswami & Middleton, 1994).
Flavonoids act as potent metal chelators and free-
radical scavengers. They are powerful chain-breaking
49
antioxidants (Santos et al., 1998). Moreover, flavonoids
are known to possess vitamin C–stabilizing and
antioxidant-dependent vitamin C–sparing activities.
They are also known to increase the absorption of
vitamin C. In addition, flavonoids are known to modify
the activities of a host of enzyme systems including pro-
tein kinase C, protein tyrosine kinase and various other
kinases, aldose reductase, myeloperoxidase, NADPH
oxidase, xanthine oxidase, phospholipase, reverse tran-
scriptase, ornithine decarboxylase, lipoxygenase, cyclo-
oxygenase, and so on (Park & Pezzuto, 2002a). Some
of these enzyme systems are critically involved in immune
function, carcinogenesis, cellular transformation, and
tumor growth and metastasis. The physiologic and
pathologic processes affected by flavonoids are diverse
and numerous and include secretion, mitogenesis, plate-
let aggregation and adhesion to endothelial surface, cell
motility, malignant cell proliferation, cancer metastasis,
and function=expression of adhesion molecules in vari-
ous mammalian cell types. The antioxidant function
and enzyme-modifying actions of flavonoids could
account for many of their pharmacological activities
(Santos et al., 1998).
Quercetin and other flavonoids are effective inhibitors
of O2  production by cells. Quercetin is a potent inhi-


bitor of human neutrophil degranulation and O2 

production, and also inhibits the phosphorylation of
neutrophil proteins accompanying neutrophil activation
by phorbol myristate acetate (Blackburn et al., 1987).
Quercetin can also suppress lipid peroxidation in
several biological systems, such as mitochondria, micro-
somes, chloroplasts, and erythrocytes. Silymarin, a
3-hydroxyflavone present in Silybum marianum Gaertne
(the European milk thistle), protects rat liver mitochon-
dria and microsomes from lipid peroxide formation
induced by Fe2 þ-ascorbate and NADPH-Fe3 þ-ADP sys-
tems (Valenzuela et al., 1987). Soybean isoflavonoids
have shown antioxidative potency and prevent peroxida-
tive hemolysis of sheep, rat, and rabbit erythrocytes
(Santos et al., 1998). Quercetin and silybin were reported
 50 T.P. Kondratyuk and J.M. Pezzuto
to exert a protective effect by preventing the decrease in
the xanthine dehydrogenase=oxygenase ratio observed
during ischemia=reperfusion in the rat (Valenzuela et
al., 1985). The enzyme xanthine oxidase, implicated in
tissue oxidative injury after ischemia=reperfusion, is a
source of reactive oxygen species that is formed from a
dehydrogenase during ischemia. The protective effect of
quercetin and silybin on the xanthine dehydrogenase=
oxidase ratio is due to inhibition of the dehydrogenase.
The bioactivity of phenolics may be related to their anti-
oxidant behavior, which is attributed to their ability to
chelate metals, inhibit lipoxygenase, and scavenge free
radicals. However, phenolics can also function as pro-
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oxidants by chelating metals in a manner that maintains
or increases their catalytic activity.
Flavonoids, especially those with catechol or pyrogal-
lol groups, obviously are prone to autoxidation reactions
(Bolton et al., 1997). The pro-oxidative and antioxidative
properties of phenolics from soybeans and other legumes
were documented by Morgan et al. (1997). The phenolic
acids and flavonoids were able to reduce ferric to ferrous
ions and were able to chelate and alter the catalytic
activity of iron. Most of the phenolics tested were also
able to inhibit the oxidation of linoleic acid micelles
For personal use only.
and ferrous ion-catalyzed oxidation of glutamine
synthase, presumably through free-radical scavenging
and removal of iron from catalytic sites via chelation.
Although phenolics inhibited oxidation in certain sys-
tems, they did not protect against all forms of oxidative
damage. Even though the phenolics chelated iron, this
metal ion was still catalytically active and able to oxidize
both deoxyribose and DNA. Pro-oxidant activity of phe-
nolics has also been observed for carnosol, carnosic acid,
quercetin, rutin, and luteolin (Morgan et al., 1997).
It was also found that pH was essential in determining
the oxidative role of phenolics. In general, a decrease in
pH increased iron-reducing activity and reduced the abil-
ity of phenolics to chelate and inhibit the catalytic
activity of iron. Increasing pH increased deoxyribose
and DNA oxidation. Inhibition of lipid oxidation was
also influenced by pH, with c-resorcyclic acid being
antioxidative at pH 5.8 and pro-oxidative at pH 7.4.
Hydrobenzoic acid was antioxidative, and apigenin-7-
glucoside was pro-oxidative at pH 7.4, yet neither had
an effect on lipid oxidation at pH 5.8. These results
suggest that the pH of biological tissues could also influ-
ence the antioxidative=pro-oxidative activity of phenolics
(Decker, 1997).
A possible mechanism of polyphenol cytotoxicity may
be related to their pro-oxidant properties (Sergediene
et al., 1999). Flavonoids autoxidize in aqueous medium
and may form highly reactive HO radicals in the pres-


ence of transition metals. In addition, polyphenols and
flavonoids may act as substrates for peroxidase and
other metalloenzymes, yielding quinone- or quinone-
methide-type pro-oxidant and=or alkylating products.
The pro-oxidant character of polyphenol cytotoxicity is
supported by the formation of activated oxygen species
during gallic acid–induced apoptosis and by the enhance-
ment of gallic and caffeic acid–induced apoptosis by non-
toxic concentrations of copper ions (Yamanaka et al.,
1997).
Catechins, such as (  )-epicatechin and (  )-epigalloca-
techin, abundant in green tea, possess the antioxidative
and pro-oxidative characteristics of Cu2 þ -induced
low-density lipoprotein (LDL) oxidation. In the
initiation phase, LDL oxidation was inhibited by
addition of catechin. In contrast, during the propa-
gation phase of LDL oxidation, catechins served as
accelerators of oxidation. Depending on redox status,
they might form reactive oxidation products such as
semiquinones and quinones and function to stimulate
oxidative reactions (Yamanaka et al., 1997).
Quercetin, a highly studied antioxidant flavonoid, has
potential to inhibit free-radical processes in cells by (a)
scavenging O2 , (b) blocking lipid peroxidation, (c)


reacting with peroxyl or lipid peroxyl radicals, (d) inhi-
biting formation of HO , and (e) chelating iron ions.


The biological effects of quercetin are believed to result
from antioxidant properties. It was demonstrated clearly
that quercetin could function both as an antioxidant and
a pro-oxidant, depending on concentration and free-
radical sources and their location in the cell.
Data on the antioxidant capability of natural polyphe-
nols allows speculation about the antioxidant activities
of tea and red wines. Green tea, which is not fermented,
is very rich in pyrogallol derivatives. It is customary to
assume these components are the dominant reason for
the very high antioxidant activity of green tea. Red
wines, during the course of aging, exhibit progressive
oxidative transformation of polyphenols and a visible
decrease in total content. Most likely, this is compen-
sated for by the elevated antioxidant activities of trans-
formation products including dimmers=oligomers. It was
recently shown that products of oxidative transformation
display a higher ability to inhibit lipid oxidation than
original polyphenols (Roginsky, 2003).
Anticarcinogenic effects of polyphenols
Carcinogenesis can be viewed as a multistage, microevo-
lutionary process. The progression of tumor formation
may be slow, often taking 10 or more years. It is
generally agreed that tumors can be derived from single
abnormal cells, and work with experimental systems
shows that carcinogenesis is divisible into three major
stages: initiation, promotion, and progression. Initiation
is a heritable aberration of a cell. Cells so initiated can
undergo transformation to malignancy if promotion
and progression follow. Initiation appears to be irrevers-
ible and can result from DNA damage. Promotion, on
 Natural product polyphenols of relevance to human health
the other hand, is affected by factors that do not alter
DNA sequences and involves the selection and clonal
expansion of initiated cells. This process is partly revers-
ible and accounts for a major portion of the lengthy
latent period of carcinogenesis. The final stage of tumor
formation is the progression of a benign growth to a
malignant neoplasm. There is loss of growth control,
an escape from the host defense mechanism, and
metastasis.
Certain initiators, such as radiation or chemical carci-
nogens, can induce the production of various free radi-
cals and subsequent DNA base sequence alteration. In
addition, cells of the immune system, such as neutrophils
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and macrophages, produce O2 and H2O2 that have


been associated with the induction of experimental can-
cers. Oxygen free-radicals and methyl radicals are known
to damage DNA. In some cases, such free radicals may
arise in reactions catalyzed by ferric and cupric ions loca-
lized in the vicinity of cellular DNA. Free radical–
mediated DNA damage can have serious consequences
on an organism unless the damage is repaired. Although
oxygen free-radical effects can lead to DNA damage,
they may also directly affect the protein components of
the DNA repair apparatus. Unrepaired DNA alterations
For personal use only.
are inherited as mutations. Phase I metabolizing enzymes
(e.g., cytochrome P450) play an important role in the
initiation stage. Through the catalytic activity of these
enzymes, a polar reactive group is added to lipophilic
carcinogens=xenobiotics to form an electrophile, which
can react with DNA. Later, the xenobiotic can be detoxi-
fied by phase II metabolizing enzymes, with sugars,
amino acids, and glutathione (Galati et al., 2000), for
example. Green and black tea extracts strongly inhibit
neoplastic transformation in mammary organ cultures
or epithelial cells (Steele et al., 2000). In animals, green
or black tea also induce the phase I metabolizing enzyme
CYP1A2.
It is generally believed that the generation of growth
promotion oxidants is a major trigger of the tumor pro-
motion and progression stages. Tumor-promoting phor-
bol esters not only can induce changes in cellular genes
leading to some of the phenotypic characteristics of
tumor cells, but they also can stimulate inflammatory
leukocytes to release superoxide. The release of super-
oxide by phagocytic cells following stimulation with
phorbol esters is proportional to their tumor promoting
activity. Low levels of both O2  and H2O2, products of


the ‘‘respiratory burst’’, can promote fibroblast growth,
possibly fibroblasts that harbor an oncogene or a
mutated protooncogene. Also, low levels of superoxide
can stimulate growth or growth responses in a variety
of cell types when added exogenously to culture medium.
In particular, these species stimulate the activation and
the translocation of protein kinase C as well as the
expression of early growth-regulated genes, such as the
protooncogenes c-fos and c-myc. Superoxide and=or
51
hydrogen peroxides might function as mitogenic stimuli
through biochemical processes common to natural
growth factors. Thus, signaling of growth responses
involving released superoxide or hydrogen peroxide
may be mediated through the oxidative modification of
components of the signal transduction pathway. It is also
possible that oxidative inactivation of serum protein
inhibitors allows proteases to remodel the cell surface,
thereby facilitating, or modulating, the action of normal
growth factors (Burdon, 1993). Nonsteroidal anti-
inflammatory drugs or COX-2 inhibitors have shown
potent chemopreventive activity in animal colorectal car-
cinogenesis models (Cuendet & Pezzuto, 2000; Krishnan
et al., 2000). Dietary polyphenolics, such as curcumin,
chlorogenic acid, caffeic acid, resveratrol, or the flavo-
noid silymarin have also been shown to prevent colon
carcinogenesis.
A final and decisive step in carcinogenesis is the
invasion and metastatic spread of the tumor to various
body spaces and cavities. This appears to be facilitated
by the activation of genes for the release of proteolytic
enzymes. Whereas high levels of immune cells appear
to favor cell killing, lower numbers of immune cells
can favor metastasis. Again, the release of superoxide
may serve to promote metastatic growth. Alternatively,
superoxide could inactivate serum antiproteases,
some of which are extremely sensitive to oxidative
inactivation (Floyd, 1990). Chemoprevention by flavo-
noids or polyphenolics could also result from tumor
cell death, apoptosis, caused by the cytotoxic effect of
flavonoids=polyphenolics. An accumulation of evidence
has shown some anticancer properties of resveratrol are
related to downregulating of the activation of NF-jB,
which contributes to the progression of the androgen-
independence of prostate cancer and increases invasive
and metastatic properties (Tsai et al., 1999). Recently,
another possible antitumor molecular mechanism of
resveratrol was shown, which involves mitogen-activated
protein kinase-mediated p53 activation and subsequent
induction of apoptosis (She et al., 2002).
Lipid peroxidation is associated with some phases of
carcinogenesis. There is increasing evidence that covalent
binding of carcinogens or toxic substances to cellular
macromolecules, particularly those carrying genetic
information, is a primary event in the initiation of carci-
nogenesis. Thus, covalent binding to macromolecules
could be the basis of many pathological changes induced
by toxic substances. The ultimate forms of xenobiotics
are believed to be reactive electrophilic metabolites,
which combine with nucleophilic groups of macromole-
cules. It is also possible that miscoding or mutagenesis
may be of minor importance in the initial events of
chemical carcinogenesis and that genetic transpositions,
including relatively large regions of the genome, may
be more relevant (O’Brien, 1994). The DNA adducts,
deoxyadenosine and deoxyguanosine, which are induced
 52 T.P. Kondratyuk and J.M. Pezzuto
by malondialdehyde, the end-product of lipid peroxi-
dation, accumulate in human breast cancer cells com-
pared to normal breast cells (Wang et al., 1996). Serum
antioxidative vitamin levels and lipid peroxidation were
compared in gastric cancer patients (Choi et al., 1999).
The level of serum ascorbic acid, a-tocopherol,
b-carotene, and retinol were assessed. The levels of
ascorbic acid in patients with gastric carcinoma were less
that one-fifth of that in the control group, and the pro-
duction of b-carotene and a-tocopherol were decreased,
as well.
In summary, current information supports the impor-
tance of polyphenols in cancer protection (Dragsted,
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2003; Rasmussen & Breinholt, 2003). A number of
studies suggest that consumption of fruits and vegeta-
bles is associated with decreased risk of colon, breast,
lung, stomach, and esophageal cancer (World Cancer
Research Fund, 1997). Nonetheless, the overall incidence
of cancers such as lung, breast, and prostate has
increased. It is reasonable to expect that the incidence
of cancer can be substantially reduced by diet modifi-
cation, regular exercise, and avoiding tobacco smoke.
For personal use only.
Cardiovascular and cerebrovascular disease
Polyphenolic compounds are generally known to possess
antioxidant properties (Tapiero et al., 2002; Bub et al.,
2003). Despite intake of a high-fat diet, the low incidence
of coronary heart disease in France—the ‘‘French Para-
dox’’—has been attributed partly to consumption of red
wine (Sun et al., 2002). Compounds such as resveratrol,
quercetin, catechin, and proanthocyanidins are enriched
in grape skins and seeds, and the ability of these
compounds to inhibit platelet aggregation and protect
low-density lipoproteins from oxidation has been
demonstrated (Xia et al., 1998). Also, the intake of flavo-
noids in relationship to cardiovascular disease has been
explored by several groups of investigators. Hertog
et al. (1997) reported that a high intake of flavonols was
associated with a decreased risk of coronary heart disease
mortality. They also reported a general decrease in mor-
tality with increasing flavonol intake. Knekt et al. (1996)
reported similar results, with a decreased risk of coron-
ary mortality associated with flavonoids. In contrast, a
U.S. study of heart disease in males 40–75 years of age
found no significant association with flavonoid intake
(Rimm et al., 1996). Finally, in a 10-year follow-up study
of more than 34,000 postmenopausal women from Iowa,
flavonoids intake was associated with a decreased risk
of heart disease. However, no association was found
between flavonoid intake and stroke mortality after 10
years of follow-up (Yochum et al., 1999).
The syndrome of ischemia =reperfusion (I=R) injury
has been characterized for the heart, brain, intestine, kid-
ney, and other organs. This phenomenon consists of a
paradoxical increase in tissue injury during the reperfu-
sion period in an organ that has sustained relatively
minor damage during a period of ischemia. It is now evi-
dent that reperfusion tissue injury is mediated through
oxidant mechanisms associated with the generation of
oxygen-based radicals. ROS have been implicated in
both the myocardial dysfunctions that are observed dur-
ing reperfusion following short periods of ischemia (the
stunned myocarium) and the irreversible injury to car-
diac myocytes that occurs during reperfusion after longer
periods of ischemia (Ferrari, 1994).
Infusions of high concentrations of the catechola-
mines epinephrine or norepinephrine into experimental
animals are known to produce myocellular mitochon-
drial swelling, myofibrillar disruption, plasma membrane
blebbing, and myocardial necrosis. It has been suggested
that these cardiotoxic effects result not from the catecho-
lamines themselves but from the production of O2 and


H2O2 formed by a complicated series of reactions during
the autoxidation of catecholamines. It was observed
that vitamin E–deficient rats were more sensitive to the
cardiotoxic effects of isoproterenol, whereas myocardial
damage induced by this synthetic catecholamine was
reduced when the diet was supplemented with vitamin
E. However, the results of studies demonstrating protec-
tion by antioxidants against catecholamine-induced
myocardial necrosis must be interpreted with caution,
as accumulation of neutrophils, a major source of oxygen
radicals, has been observed in such models (Singal et al.,
1982).
Carvedilol, a potent antioxidant, prevents the lipoper-
oxidation of mitochondrial membranes, which suggests
a strong contribution to the known cardioprotective
activity of this compound through protection of mito-
chondrial function (Moreno et al., 1998). A similar cardi-
oprotective benefit is achieved by agents and antioxidant
enzymes that scavenge hydroxyl radicals (or reduce their
formation), but not agents that reduce superoxide anion
production. Some examples of compounds of plant ori-
gin that have shown protective effects against ischemic
injury are procyanidine from Vitis vinifera L. (Facino
et al., 1996), resveratrol from red wine (Ray, 1999),
and ginseng extract (Facino et al., 1999).
The ability of polyphenolic compounds to offer cardio-
vascular system protection has also stimulated efforts to
investigate whether these compounds may offer neuro-
protective effects. Few studies have explored flavonoid
intake and risk of stroke. An inverse association has been
shown with increasing dietary quercetin consumption.
Tea consumption, which comprised the major source of
flavonoids intake, was associated with decreased risk of
stroke (Keli et al., 1996). Knekt et al. (2000) showed that
quercetin intake is not associated with cerebrovascular
disease.
Lipid-peroxidation of biological membranes gives
rise to degeneration of synapses and neurons and may
 Natural product polyphenols of relevance to human health
be observed in stroke or neuronal disorders such as
Alzheimer, Parkinson, and Huntington diseases.
Oxidative stress and damage are accepted features of
neural degeneration. The pathological presentation of
Alzheimer disease, the leading cause of senile dementia,
involves regionalized neuronal death and accumulation
of intraneuronal and extracellular lesions (Smith et al.,
1997). 4-Hydroxynonenal mediates oxidation-induced
impairment of glutamate transport and mitochondrial
function in synapses (Keller et al., 1997) Amyloid b-
protein may be related to modulation of membrane lipid
peroxidation. Amyloid b-protein fragment 25–35 [A-beta
(25–35)] inhibits lipid peroxidation at low concentrations
Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
as a result of physicochemical interactions with the
membrane lipid layer (Walter et al., 1997). Further, there
is close association between increased levels of the
antioxidant enzymes superoxide dismutase and heme
oxygenase-1 and cytoskeleton abnormalities found in
Alzheimer disease (Smith & Perry, 1998).
More studies will be needed to test specific effects of
individual polyphenols and understand their molecular
mechanism of action at the subcellular level during heart
and neuronal diseases.
For personal use only.
Discovery of new chemopreventive
agents from plants
Botanicals have been used for the treatment of various
human diseases throughout history. In addition to treat-
ment, however, botanicals can play a role in disease pre-
vention. There is growing interest in medicinal botanicals
as part of complementary medicine in the United States.
Approximately 40% of Americans use alternative rem-
edies, including herbal medicine, for disease prevention
and therapy (Eisenberg et al., 1998). The high cost, side
effects, and therapeutic limitations of conventional med-
ications are key factors that are driving the revival of her-
bal remedies. People can easily obtain herbal medicines
or botanical supplements on an over-the-counter basis
and feel comfortable consuming these products, even
though safety and efficacy are generally not established
on a rigorous scientific basis. This may be problematic
due to factors such as drug-drug interactions (Go et al.,
2001; von Gyuenigen & Hopkins, 2000). On the other
hand, these preparations may mediate beneficial
responses, either due to active chemical constituents or
placebo effects.
During the past several years, we have instituted
a multidisciplinary project wherein plant materials
procured from throughout the world are used for
activity-guided fractionation schemes that yield novel
and otherwise unpredictable chemical entities with desir-
able biological potential. The overall experimental
approach for procuring cancer chemopreventive agents
in this manner has been described in the literature
53
(Pezzuto, 1997; Pezzuto et al., 1998, 2004). In the field
of natural product drug discovery, a time-tested method
to enhance success is bioassay-guided fractionation. How-
ever, definition of the ‘‘best’’ bioassay system for monitor-
ing cancer chemoprevention remains subjective. Use of
in vitro assays enables the identification of a suitable num-
ber of lead starting materials, and bioassay-directed iso-
lation facilitates the procurement of active agents from a
reasonable number of sources in a finite period of time.
There are many assays that can be used in drug discovery
process. Several of them, which have been extensively used
in our laboratory, are presented below.
Induction of quinone reductase activity utilizing
cultured hepa 1c1c7 cells
Induction of phase II drug-metabolizing enzymes, such as
glutathione S-transferase (GST) and quinone reductase
(QR), is an important mechanism of cancer chemopre-
vention. In searching for novel cancer chemopreventive
agents, we have used a rapid, sensitive QR assay to iden-
tify potential detoxification enzyme inducers (Talalay
et al., 1995; Kang & Pezzuto, 2004). In order to under-
stand the possible mechanism of induction, we have also
employed a series of methods to further investigate induc-
tion patterns, protein expression, and mRNA expression,
using transient transfection, Western blotting, Northern
blotting, and reverse transcription-polymerase chain
reaction (RT-PCR) techniques (Gerhäuser et al., 1997a;
Song et al., 1999). Potent enzyme inducers are evaluated
for potential to inhibit carcinogen-induced lesion forma-
tion in mouse mammary organ culture (MMOC) (Mehta
et al., 1995). Chemopreventive activities of promising
agents are studied for their inhibitory effects in
animal models, such as the two-stage mouse skin or the
rat mammary gland carcinogenesis models.
Antioxidant activity
Despite the abundance of published experimental
approaches devoted to the antioxidant activity of poly-
phenols, only a few studies present reproducible infor-
mation. Because reactive oxygen radicals play an
important role in carcinogenesis, antioxidants present
in consumable fruits, vegetables, and beverages have
received considerable attention as cancer chemopreven-
tive agents (Lee et al., 1998, 1999b). In order to identify
antioxidants in plant extracts, activity has been assessed
by determining scavenging activity with stable 2,2-
diphenyl-1-picryhydrazyl (DPPH) free radicals (Fujita
et al., 1988), inhibition of 12-O-tetradecanoyl-13-phorbol
acetate (TPA)-induced free-radical formation with cul-
tured HL-60 cells (Sharma et al., 1994), and inhibition
of superoxide anion production in xanthine=xanthine
oxidase systems (Sheu et al., 1998).
 54 T.P. Kondratyuk and J.M. Pezzuto
HL-60 cell differentiation
The HL-60 cell line has been used to provide a unique
in vitro model system for studying the cellular and mole-
cular events involved in the proliferation and differen-
tiation of normal and leukemic cells. Terminal
differentiation of human promyelocytic leukemia cells
can be induced by a variety of chemical agents, and this
process can be monitored readily by the generation of
morphologically, histochemically, and functionally
mature granulocytes and monocytes=macrophages (Suh
et al., 1995; Mata-Greenwood et al., 2001). Induction
of terminal differentiation is of interest for the therapy
Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
of leukemia, and human promyelocytic leukemia cell
lines serve as convenient in vitro models to study differen-
tiation. The usefulness of active compounds could be
more far reaching, however, as substances such as vit-
amin A and D metabolites facilitate cell differentiation.
Inhibition of cyclooxygenase
Cyclooxygenase, a key enzyme in prostaglandin (PG)
biosynthesis, catalyzes oxygenation of arachidonic acid
to prostaglandin G2 (PGG2) and reduction of PGG2 to
For personal use only.
prostaglandin H2 (PGH2), an immediate precursor for
production of eicosanoids. Arachidonic acid metabolites
derived from PGH2 are important mediators of inflam-
matory responses, immunological effects, and tumor
development. Several epidemiological studies have
demonstrated that nonsteroidal anti-inflammatory drugs
(NSAIDs) have cancer chemopreventive and tumor
inhibitory effects in the human colon (Logan et al.,
1993). Overproduction of PGs may influence tumor
growth, carcinogen metabolism, and metastatic potential
in human beings and experimental animals (Cuendet &
Pezzuto, 2000). COX-2 is expressed in colorectal aden-
oma and carcinomas. Inhibition of COX-2 has been
shown to decrease the incidence of carcinogen-induced
neoplasia in rats and to reduce the incidence of adeno-
mas in murine models. Several COX-2 inhibitors, with
the potential for less toxicity than that associate with tra-
ditional NSAIDs, are currently available (Lynch, 2001).
Anti-estrogenic activity
For the evaluation of anti-estrogenic activities, we have
employed an Ishikawa cell-based assay and an assay to
determine inhibition of aromatase. The Ishikawa cell
system has been employed to determine antiestrogenic
activities (Pisha & Pezzuto, 1997). The Ishikawa cell line
is a stable human endometrial carcinoma that expresses
an estrogen-dependent alkaline phosphatase. Induction
of this enzyme can be estimated kinetically with a micro-
titer plate reader. Thus, the assay is adaptable to large-
scale screening using a 96-well plate format. In addition,
the aromatase enzyme complex is responsible for the
conversion of androgens to estrogens (Grubjesic et al.,
2002). In the case of estrogen-dependent tumorigenesis,
aromatase expression may be an important factor in
the regulation of tumor growth. Therefore, inhibition
of aromatase, the terminal step in estrogen biosynthesis,
provides a method for treating hormone-dependent
breast cancer (Santen & Harvey, 1999). Third-generation
aromatase inhibitors have replaced megestrol acetate as
second-line hormonal therapy in advanced breast cancer,
and large clinical trials are designed to establish their
efficacy, relative to tamoxifen (Ingle, 2001).
Unfortunately, no short-term in vitro systems are cap-
able of fully representing the physiological complexity of
mammals. Thus, to help overcome this disadvantage, we
have assembled a panel of bioassay procedures that
reflect various stages of carcinogenesis. The composition
of the panel varies over time but generally includes assays
that are suitable for monitoring inhibition of carcinogen-
esis at the stages of initiation (e.g., antioxidant activity
and induction of quinone reductase activity in cell cul-
ture), promotion (e.g., inhibition of TPA-induced ODC
activity in cell culture and inhibition of cyclooxygenase
activity), and progression (e.g., induction of cell differen-
tiation and anti-estrogenic activity). More than 15,000
tests have been performed, and the number of plants
characterized as ‘‘active’’ is in the range of 3% of the
total. In general, these ‘‘active’’ plant extracts are tested
in a secondary model of greater physiological com-
plexity. One example is a test to assess potential to
inhibit carcinogen-induced preneoplastic lesion for-
mation in the mouse mammary organ culture model
(Mehta & Pezzuto, 2002; Pezzuto, 1997; Pezzuto et al.,
1998, 2004). Active leads in the secondary model are sub-
jected to bioassay-guided fractionation using an in vitro
system as a monitor, and active isolates are considered
for evaluation in full-term animal studies. Examples of
edible plant isolates are summarized in Table 1.
As compared with edible plants that are used in daily
life as a food, we consider medicinal plants as those with
pharmacological activities to treat disease. There have
been many scientific reports describing diverse clinical
uses and bioactive components of medicinal plants. Ani-
mal trials to evaluate the chemopreventive activity of
some well-known traditional medicinal plants are sum-
marized in Table 2 (Park & Pezzuto, 2002b). Various
groups of compounds have been classified as cancer che-
mopreventive agents, largely based on the results of ani-
mal studies and epidemiological data. A series of studies
to evaluate the chemopreventive activity of medicinal
plants has been performed in Japan. The approach has
typically involved primary screening of many natural
products followed by a two-stage carcinogenesis assay
to assess inhibitory effects on mouse skin tumors induced
by the DMBA=TPA protocol. As a primary screen, com-
pounds have been tested for their inhibitory effects on
Epstein-Barr virus early antigen (EBV-EA) induction
 Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
For personal use only.

Table 1. Potential chemopreventive agents isolated from edible plants.

Latin name Compound Activity References

Broussonetia papyrifera 30 -[c-Hydroxymethyl-(E)-c-methylallyl]-2,4,20 ,40 - AE Lee et al. (2001c)

(L.) Venten
Casimiroa edulis Llav.
et Lex
Cerbera manghas L.
Cotinus coggygria
Scop.
Eugenia sandwicensis
Gray
Isodon excisus var
coreanus Nakai
Physalis philadelphica
Lam.
Tephrosia purpurea
Pers.
Thuja occidentalis L.
AE, antiestogenic activity; AO, antioxidant activity; ODC, ornithine decarboxylase inhibitor; QR, quinone reductase inducer; HL-60, HL-60 cell differentiation inducer;
tetrahydroxychalcone 110 -O-coumarate
(2S)-20 ,40 -Dihydroxy-200 (1-hydroxy-1-methylethyl)-
dihydrofuro[2,3-h]flavanone
Isolicoflavonone
(2S)-Abyssinone II
Zapotin HL-60 Mata-Greenwood et al. (2001)
20 ,5,6-Trimethoxyflavone
()-14-Hydroxy-3b-(3-O-methyl-6-deoxy-a-l-rhamnosyl)-11a, AE Chang et al. (2000a)
12a-epoxy-(5b, 14b,17bH)–card-20(22)-enolide
()-14-Hydroxy-3b-(3-O-methyl-6-deoxy-a-l-
glucopyranosyl)-11a, 12a-epoxy-(5b, 14b,17bH)–card-
20(22)-enolide
()-17b-Neriifolin
Disulfuretin, sulfuretin, sulfurein AO Westenburg et al. (2000)
Gallic acid, methyl gallate
Pentagalloyl glucose
Gallic acid AO Gu et al. (2001)
3b-trans-p-coumaroyloxy-2a,23-dihydroxyolean- MMOC
12-en-28-oic acid
Inflexin AE Jeong et al. (2000)
Ursolic acid
Ursolic acid 3-O-acetate
2,3-Dihydro-3-methoxywithaphysacarpin QR Kennelly et al. (1997)
Withaphysacarpin
24,25-Dihydrowithanolide
Philadelphicalactone A=B QR Su et al. (2002b)
Ixocarpalactone A=B
18-Hydroxywithanolide D
Withanone
7,40 -Dihydroxy-30 ,50 -dimethoxyisoflavone QR Chang et al. (1997)
(þ)-Tephropurpurin, (þ)-purpurin
Pongamol, lanceolatin
()-Maackiain, ()-medicarpin
()-3-Hydroxy-4-methoxy-8,9-methylene- dioxypterocarpan
(þ)-7-Oxo-13-epi-pimara-14,15-dien-18-oic acid ODC Chang et al. (2000b)
(þ)-7-Oxo-13-epi-pimara-8,15-dien-18-oic acid
(þ)-Isopimaric acid
(1S,2S,3R)-(þ)-Isopicryodeoxypodophyllotoxin
()-Deoxypodophyllotoxin
()-Deoxypodorhizone

55
MMOC, mouse mammary organ culture.
 Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
For personal use only.

56
Table 2. Chemopreventive medicinal plants reported in animal carcinogenesis models.

Latin name (common name) Active component=biological activity References

Ajuga decumbens Thunb. 8-Acetylharpagide Takasaki et al. (1999b);

Inhibition of two stage mouse skin carcinogenesis and Konoshima et al. (2000)
mouse pulmonary tumor
Aloe barbadensis Miller Polysaccharide Kim et al. (1999);
Inhibition of BP-DNA adduct formation Kim and Lee (1997)
Inhibition of ODC activity
Inhibition of tyrosine kinase activity
Apium graveolens L. (celery seed oil) p-Mentha-2,8-dien-1-ol, 3-n-butyl phthalide, sedanolide Zheng et al. (1993)
Induce GST
Inhibition of BP-induced stomach cancer
Asteracantha longifolia Aees. Methanol extract of seed Ahmed et al. (2001)
Inhibition of hepatocarcinogenesis in Wistar rats
Increase GPx and CAT, ODC
Bolbostemma paniculatum (Maxim.) Franquet Tubeimoside I and tubeimoside III Yu et al. (1995)
Antitumor promotion activity against TPA
Calophyllum inophyllum L. Calocoumarin-A Itoigawa et al. (2001)
Inhibition of two-stage mouse skin carcinogenesis
Chelidonium majis L. Inhibition of MNNG-induced glandular stomach carcinogenesis Kim et al. (1997)
Coleogyne ramosissima Torr. Polyphenols Ito et al. (1999)
Cowania mexicana D. Don. Inhibition of two stage mouse skin carcinogenesis
Coptis japonica (Thunb.) Makino Inhibition of AOM-induced ACF Fukutake et al. (1998)
Eucalyptus grandis Hill ex Maiden Euglobal-G1(phloroglucinol-monoterpene) Takasaki et al. (2000);
Inhibition of two-stage mouse skin carcinogensis Takasaki et al. (1995)
Inhibition of 4-NQO–induced mouse pulmonary tumor
Ferula narthex Boiss. Antioxidant and inhibition ODC activity Saleem et al. (2001f)
Ganoderma lucidum Karst. GST induction Kim et al. (1999);
Inhibition of lipid peroxidation and oxidative DNA damage Lee et al. (2001)
Hibiscus sabdariffa L. Antimutagenicity in Salmonella mutation assay Chewonarin et al. (1999)
Ocimum sanctum L. Extract of leaves Banerjee et al. (1996)
Induce GST
Picrorhiza kurroa Royle ex Benth. Picroliv (iridoid glycoside mixture) Rajeshkumar and Kuttan (2001)
Inhibition of 3-MC-induced sarcoma and DMBA-induced papilloma
Scutellaria baicalensis Georgi Inhibition of AOM-induced ACF Fukutake et al. (1998)
Semecarpus anacardium L. Antioxidant Premalatha and Sachdanandam (1999)
Taraxacum japonicum Koidz Inhibition of two-stage mouse skin carcinogenesis Takasaki et al. (1999a)
Wistaria brachybotrys Sieb et Zucc. Pendulon Konoshima et al. (1997)
Inhibition of two-stage mouse skin carcinogenesis

ACF, aberrant crypt foci; AOM, azoxymethane; BP CAT, catalase; GPx, glutathione peroxidase; GST, glutathione transferase; 23-MC, 3-methylcholanthrene; MNNG,
N-methyl-N0 -nitro-N-nitrosoguanidine; 4-NQO, 4-nitroquinoline-N-oxide; ODC, ornithine decarboxylase.
 Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
For personal use only.

Table 3. Potential chemopreventive agents isolated from medicinal plants.

Latin name Compound Activity References

Acnistus arborescens (L.) Schltdl. 7b-Acetoxywithanolide D QR Minguzzi et al. (2002)

7b,16a-Diacetoxywithanolide D
4-Deoxy-7b,16a-diacetoxywithanolide D
Aglaia ponapensis Kanehira Desmethylrocaglamide HL-60 Mata-Greenwood et al. (2001)
Aiphanes aculeate Willd. Aiphanol COX Lee et al. (2001d)
Antirhea acutata (DC) Urb. (6S)-Hydroxy-29-nor-3,4-seco-cycloart-4(30),24-dien-3-oic acid COX AO Lee et al. (2001e)
8-[1-(3,4-Dihydroxyphenyl)-3-methoxy-3-oxopropyl]epicatechin
Brassica campestris L. Brassinin QR Gerhäuser et al. (1997a)
Brucea javanica (L.) Merr. Yadanziolide S HL-60 Su et al. (2002a)
Flazin COX
Bruceine MMOC
Yadanxioside
Bruceoside A
Yadanzioside F
Guaiacylglycerol-b-O-60 -(2-methoxy)cinnamyl alcohol HL-60 Luyengi et al. (1996)
Brusatol
Dehydrobrusato
Yadanziolide C
Blumenol A
Chorizanthe diffusa Benth. 5,8,30 ,40 ,50 -Pentahydroxy-3,7-dimethylflavone AO Chung et al. (1999)
5,7,30 ,40 -Tetrahydroxy-3-methylflavone MMOC
5,8,30 ,40 -Tetrahydroxy-3,7-dimethylflavone
Quercetin
300 -O-Acetylquercetin
Daphniphyllum calycinum Benth. 5,6,7,40 -Tetrahydroxyflavonol-3-O-rutinoside AO Gamez et al. (1998)
3-O-Neopesperidoside
Mundulea sericea (Willd.) A.Chev. Deguelin ODC Udeani et al. (1997);
Gerhäuser et al. (1997b)
Munetone ODC Lee et al. (1999a)
Pachysandra procumbens Michx. ()-Pachyaximine A AE Chang et al. (1998)
(þ)-Pachysamine B
(þ)-(20S)-S-(Benzoylamino)-20-
(dimethylamino-5a-pregn-2-en-4b-yl acetate
Petiveria alliacea L. Dibenzyltridulfide HL-60 Mata-Greenwood et al. (2001)
2-[(Phenylmethylditho)ethanol

AO, antioxidant activity; AE, anti-estrogenic activity; COX, cyclooxygenase inhibitor; ODC, ornithine decarboxylase inhibitor; QR, quinone reductase inducer; HL-60, HL-60
cell differentiation inducer; MMOC, mouse mammary organ culture.

57
 58 T.P. Kondratyuk and J.M. Pezzuto
by treatment of Raji cell with a tumor promoter. Using a
battery of chemopreventive assay systems, we have
reported various candidate agents. Some chemopreven-
tive agents from plants are summarized in Table 3.
At present, brassinin (Mehta et al., 1995), sulfora-
phane and sulforamate (Gerhäuser et al., 1997a), 40 -
bromoflavone (Song et al., 1999), withanolides (strong
inducers of phase II drug-metabolizing enzymes) (Suh
et al., 1995), deguelin (strong ODC inhibitor) (Kennelly
et al., 1997), brusatol (HL-60 cell differentiation)
(Mata-Greenwood et al., 2001), two aromatase inhibitors
(Talalay et al., 1995), and resveratrol (Bhat et al., 2001;
Bhat & Pezzuto, 2002) are undergoing further investi-
Pharmaceutical Biology Downloaded from informahealthcare.com by University of Alberta on 09/01/13
gation as lead compounds. Clinical trials are anticipated
in due course.
Conclusions
The results of studies outlined in this review provide a
solid rationale for improvements in dietary habits, with
special emphasis on enhanced consumption of fruits
and vegetables. As described herein, there is substantial
merit in this approach, as many fruits and vegetables
For personal use only.
have been found to contain polyphenols—active cancer
chemopreventive agents.
In the past few years, proof-of-principle has been
accomplished for cancer chemoprevention, and two
agents, tamoxifen and celecoxib, are available as pre-
scription drugs for the prevention of breast cancer and
familial polyposis, respectively. These are tremendous
breakthroughs. Nonetheless, a great deal of additional
work is required. For example, dilemmas such as the
near 100% probability of developing prostate cancer
given a sufficient lifespan, and the lack of effectiveness
of agents such as tamoxifen against estrogen receptor–
negative breast cancer, need to be recognized. Some
large-scale clinical trials are ongoing, but new cancer
chemopreventive agents are still required, and plant
materials are a promising source for the identification
of active lead compounds. Certainly, to move from a
crude starting material such as a plant extract to a pre-
scription drug proven effective as a cancer chemopreven-
tive agent is a long and arduous process. Nonetheless, it
is necessary to bear in mind the significance of the task.
The morbidity and mortality associated with metastatic
cancer should be of concern for every living human
being; no therapeutic option offers greater promise than
cancer chemoprevention. Having the potential of
administering an effective agent through the diet or as
a prescription drug is a major advantage. This flexibility
offers great hope for making significant progress in the
near future.
Human trials on the biological effects of polyphenols
have been limited. One problem is that it is difficult to
determine which polyphenols are the most protective
against cancers or cardiovascular diseases. Certain poly-
phenols inhibit COX, LOX, and phospholipase A2
activities, and they are considered potentially beneficial.
Verification of the physiological relevance of such activi-
ties and the development of reliable methods to measure
individual phenolic compounds in body fluids and tissues
are necessary steps. Some polyphenolic compounds have
been considered as chemopreventive agents on the basis
of their ability to modulate signal transduction path-
ways, cell proliferation, and apoptosis. It is important
to consider the concentrations of test agents used in these
studies and assess the ability to conduct investigations
needed in terms of comparison of the effective concen-
tration on cell lines to attain comparable levels in ani-
mals and humans. It is also important to take into
account the dose and mode of administering polyphenols
when interpreting the results of the animal experiments.
There is clearly a large amount of future research that
remains to be done, but polyphenols offer great hope
for the prevention of human disease.
Acknowledgments
Experimental work in the laboratory of the authors in
the area of cancer chemoprevention is supported by
grant no. P01 CA48112 awarded by the National Cancer
Institute. Additional cancer chemopreventive testing is
conducted under NCI contracts N01-CN-05024 (WS
no. 73), N01-CN-05124 (WS no. 79), N01-CN-15017-44
(WS no. 80), and N01-CN-15017-44 (WS no. 85).
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