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URRENT 
O
 NCOLOGY 
 —
OLUME 
13, N 
UMBER
3
99
NATURAL HEALTH PRODUCTS FOR ANGIOGENESIS INHIBITION
Natural health products thatinhibit angiogenesis: a potentialsource for investigational newagents to treat cancer—Part2
S.M.Sagar
 MD
 ,* D.Yance
 MH 
 ,
and  R.K.Wong
 MD
*
 
R E V I E W A R T I C L E
ABSTRACT
The herbalist has access to hundreds of years of ob-servational data on the anticancer activity of manyherbs. Laboratory studies are expanding the clinicalknowledge that is already documented in traditionaltexts. The herbs that are traditionally used for anti-cancer treatment and that are anti-angiogenic throughmultiple interdependent processes (including effectson gene expression, signal processing, and enzymeactivities) include
 Artemisia annua
(Chinese worm-wood),
Viscum album
(European mistletoe),
Curcumalonga
(curcumin),
Scutellaria baicalensis
(Chineseskullcap), resveratrol and proanthocyanidin (grapeseed extract),
 Magnolia officinalis
(Chinese magno-lia tree),
Camellia sinensis
(green tea),
Ginkgo biloba,
quercetin,
Poria cocos,
 
 Zingiber officinalis
(ginger),Panax ginseng,
 Rabdosia rubescens
hora (Rabdosia),and Chinese destagnation herbs. Natural health prod-ucts target molecular pathways other than angiogen-esis, including epidermal growth factor receptor, the
 HER2
 / 
neu
gene, the cyclo-oxygenase-2 enzyme, thenuclear factor kappa-B transcription factor, the pro-tein kinases, the Bcl-2 protein, and coagulation path-ways. Quality assurance of appropriate extracts isessential prior to embarking upon clinical trials. Moredata are required on dose–response, appropriate com-binations, and potential toxicities. Given the multipleeffects of these agents, their future use for cancertherapy probably lies in synergistic combinations.During active cancer therapy they should generallybe evaluated in combination with chemotherapy andradiation. In this role, they act as modifiers of bio-logic response or as adaptogens, potentially enhanc-ing the efficacy of the conventional therapies orreducing toxicity. Their effectiveness may be in-creased when multiple agents are used in optimalcombinations. New designs for trials to demonstrateactivity in human subjects are required. Althoughcontrolled trials may be preferable, smaller studieswith appropriate endpoints and surrogate markers foranti-angiogenic response could help to prioritizeagents for larger, resource-intensive phase
III
trials.
KEY WORDS
Angiogenesis, anti-angiogenic, natural health prod-ucts, herbal medicine, anticancer, clinical trials, inte-grative, molecular biology
1. INTRODUCTION
The biochemical signalling pathways of angiogen-esis form a complex, interconnected web. Inhibitionof one part of the web may result in compensationthrough another pathway. Because botanicals con-tain a variety of organic chemical complexes, theyusually act on multiple targets. A potential advan-tage of phytochemicals is that they may act throughmultiple pathways and reduce the development of resistance by cancer cells. This model of pharmacog-nosy recognizes the advantage of administering thewhole plant product to maximize activity. Over-ex-traction of a specific chemical constituent may re-move this therapeutic gain. The challenge for modernpharmacognosy is to ensure that the optimum mix-ture of chemical constituents is maintained when aproduct is purified. Usually, such assurance will re-quire a combination of chemical and biologic assays.The additional anticancer properties of some anti-angiogenic botanicals are briefly discussed here. Theirproperties may affect various biochemical pathwaysthat indirectly influence angiogenesis. Traditionalpractice has been to combine multiple natural healthproducts, and scientifically, such combination mayprovide a therapeutic advantage.
2. MULTISTEP ACTIVITY OFPHYTOCHEMICAL COMPLEXES DERIVEDFROM HERBS
2.1Targeting Alternative Angiogenesis Pathways
The adipocytokines—polypeptides produced byadipocytes—have autocrine, paracrine, and endocrineactivities, and are associated with obesity,hyperinsulinemia, and chronic vascular disease as
 
100
URRENT 
O
 NCOLOGY 
 —
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13, N 
UMBER
3
SAGAR
et al.
well as with the development of cancer
1
. Theadipocytokines include vascular endothelial growthfactor (
VEGF
), hepatocyte growth factor (
HGF
), leptin,tumour necrosis factor alpha (
TNF
α
), heparin-bind-ing epidermal growth factor, insulin-like growth fac-tor, and interleukin-6 (IL-6). All can promoteangiogenesis.Curcumin (from turmeric) and epigallocatechin-3 gallate (
EGCG
, from green tea) can inhibit aminopep-tidase-N (CD13), a member of the matrixmetalloproteinase family that is implicated in theangiogenic switch process
2–4
. Curcumin and
EGCG
canalso interfere with the expression of 
VEGF
by suppress-ing a series of activities that promote angiogenesis.These angiogenic pathways include production of transforming growth factor beta (
TGF
β
), amplificationof cyclooxygenase-2 (
COX
-2) and epidermal growthfactor receptor (
EGFR
), aberrant expression of 
src
, andamplification of nuclear factor kappa-B (
NF
-
κ 
B
) sig-nalling. Curcumin, grape seed extract, and green teaconstituents may also interfere with endothelial cellfunction by inhibiting the engagement of specificintegrins
5,6
. These phytochemicals interact at mul-tiple levels to suppress the inflammatory, hyperpro-liferative and transformative processes that constitutecarcinogenesis.
2.2Targeting EGFR (
 HER1
)
In many human tumours,
EGFR
is overexpressed. Suchoverexpression is associated with more aggressivedisease, relative resistance to cytotoxic chemotherapy,and a poorer prognosis. Activity of 
EGFR
induces an-giogenesis
7
. Blockade of 
EGFR
reduces angiogenesisand cell proliferation
8
. Monoclonal antibodies havebeen developed to block the receptor or the linkedintracellular signalling system
9–15
.Epidermal growth factor (
EGF
) stimulates uroki-nase-type plasminogen activator (u
PA
) expression,which can promote angiogenesis. Genistein (anisoflavone constituent of soy) and curcumin (a con-stituent of turmeric) both inhibit the effects of 
EGF16
.In cell cultures, genistein and curcumin inhibit
EGF
-stimulated urokinase production and phosphorylationof 
EGFR
. Both botanicals also inhibit protein tyrosinekinases, which could stimulate the enhancement of u
PA
levels induced by
TGF
β
17
.Other natural health products that can block ac-tivity of 
EGFR
include resveratrol
18
and quercetin
19–21
.
2.3Targeting
 HER2
 / 
 neu
The
 HER2
 / 
neu
gene (formerly known as c-
erbB
-2)is amplified in more than 30% of patients with breastcancer. It is linked to highly aggressive tumours witha poorer prognosis. Overexpression of 
 HER2
is alsoseen in a significant proportion of patients with othercancer types, including non-small-cell lung cancer,ovarian cancer, prostate cancer, and gastric cancer,in which it may predict a worse outcome
22–26
. Am-plification of the
 HER2
gene correlates with higherlevels of angiogenesis
27
.Herceptin (Genentech, San Francisco, CA,U.S.A.) is a drug that inhibits
 HER2
 / 
neu
. It is usu-ally administered adjunctively with cytotoxic chemo-therapy. The activity of Herceptin may be furtherenhanced by oleic acid
28
. Emodin, a natural constitu-ent of 
Polygonum multiflorum
and aloe, inhibits
 HER2
 / 
neu
expression and is toxic against cancer cells,but nontoxic for normal cells
29
.
2.4Targeting Inflammatory Pathways: COX-2 andNF-
κ κ κ κ κ 
B
Prostaglandins are autacoids derived from arachidonicacid via the
COX
enzymes. They include prostacyclin,thromboxane, and prostaglandinE types1–3. A rolefor arachidonic acid–derived prostaglandins in theprocess of angiogenesis is now established through
invitro
assays. ProstaglandinE2 is a potent inducer of angiogenesis. A correlation exists between
COX
-2 ex-pression and angiogenesis
30
. Neovascularization isblocked by
COX
-2 antagonists
31–36
.The
COX
-2 and lipoxygenase (
LOX
-5) products of omega-6 fatty acid metabolism may exert stimula-tory effects on cancer progression including angio-genesis. The omega-3 fatty acids and somepharmacologic inhibitors of eicosanoid biosynthesisantagonize these effects
37–41
. Large amounts of omega-3 fatty acids [eicosapentaenoic acid (
EPA
) anddocosahexaenoic acid] are found in cold-water fishoils. Liquorice contains glycyrrhizic acid andpolyphenols that inhibit
COX
-2,
LOX
-5, and proteinkinaseC (
PKC
) and also downregulate
EGF42
.The
NF
-
κ 
B
family consists of closely related pro-tein dimers that bind to a common sequence motif in
DNA
called the
κ 
B
site. The
NF
-
κ 
B
inducible transcrip-tion factor is increased in tissue inflammation, cellproliferation, and cancers. Nuclear factor kappaBinduces overactivation of the
COX
enzymes and isassociated with increased angiogenesis
43–46
.The
COX
enzymes are expressed in most normaltissues. The
COX
-1 enzyme synthesizes non-inflam-matory prostaglandins such as prostaglandinE1. Incontrast,
COX
-2 is amplified as part of the inflamma-tory response and produces prostaglandins such asprostaglandinE2, which may induce uncontrolled cellproliferation and carcinogenesis.Nuclear factor kappaB may be amplified bygrowth factors, including
TGF
β
and basic fibroblastgrowth factor. Besides
NF
-
κ 
B
, other transcription fac-tors, such as activator protein-1 (
AP
-1) and IL-6, canstimulate
COX
-2 transcription. Activator protein-1 alsopromotes the metastatic phase of tumour cells.
Angiogenesis mediated by
COX
-2 also has a role inthe progression of pre-neoplastic lesions to the inva-sive phenotype
47–50
. Conventional cancer therapies—such as radiation, surgery, and chemotherapy—may
 
URRENT 
O
 NCOLOGY 
 —
OLUME 
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UMBER
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NATURAL HEALTH PRODUCTS FOR ANGIOGENESIS INHIBITION
induce
COX
-2 amplification as part of the inflammatoryresponse
51
. The significance of this induction is unclear,but it could hypothetically reduce therapeutic gain.
Several phytochemical derivatives are potent in-hibitors of 
NF
-
κ 
B
. These include resveratrol, piceatan-nol, curcumin,
EGCG
, 6-gingerol (ginger), ursolic acid(holy basil), and ginseng
52–56
. Many botanical
COX
-2inhibiting agents block the amplified activity of thetranscription factor
NF
-
κ 
B
without affecting its nor-mal function.A variety of natural health products can specifi-cally inhibit the
COX
-2 enzyme and could play a rolein reducing tissue toxicity and improving tumourcontrol when used alongside therapies such as radio-therapy, chemotherapy, and surgery (Table
I
)
57
. Abotanical that protects an organism from the adverseeffects of an intervention is termed an adaptogen.Panax ginseng and curcumin are adaptogens that in-hibit
COX
-2 and that have anti-angiogenic activityderived through the inactivation of 
NF
-
κ 
B55,58–61
.
2.5Targeting Protein Kinases
Oncogenes that encode protein kinases may contrib-ute to the development of cancer. In normal cells,protein kinases are involved in signals between thecell membrane and the nucleus, regulating progres-sion through the cell cycle. Protein kinases controlthese processes by activating other messenger pro-teins that can influence cell proliferation.Mutated kinase genes have been found in a num-ber of malignancies, including chronic myelogenousleukemia and breast and bladder cancers. The mu-tated kinases can contribute to the development of cancer. In many tumour cells, protein kinases arepermanently turned on, forcing the cell into constantdivision. Examples of abnormal kinases are the
abl,src,
and cyclin-dependent kinases. The kinases maybe amplified or permanently switched on by muta-tions in the control regions of their genes. A com-monly overproduced kinase in cancer is
EGFR
.Numerous phytochemicals are reported to inter-fere with cell signalling and may reverse the adverseeffects of protein kinase overactivity. Some botani-cals with
COX
-2 inhibitory activity target the intracel-lular signalling molecules
62,63
. Inhibition of specificprotein kinases suppresses angiogenesis
64–69
.Carnosol and ursolic acid are compounds foundin
Ocimum sanctum
(holy basil) and
 Rosmarinus offi-cinalis
(rosemary)
70
. They inhibit the activity of thetyrosine kinases and ornithine decarboxylase
71
.Carnosol also reduces
NF
-
κ 
B72
and the anti-apoptoticprotein Bcl-2
73
. Genistein and daidzein (isoflavonesfound in soy) are specific inhibitors of tyrosinekinases
74
.Many phytochemicals appear to selectively reactwith the regulatory centre of 
PKC
. Curcumin, vitaminE,green tea (catechins), resveratrol,
Ganodermalucidum,
and liquorice can inhibit
PKC
activity
42,75–77
.
2.6Targeting the Bcl-2 Protein
The signalling protein Bcl-2 plays a key role in theprocess of controlled cell death called apoptosis, whichis necessary to eliminate aged or damaged cells. TheBcl-2 protein is normally found in the mitochondrialmembrane, where it regulates the release of cyto-chromeC. The latter protein can trigger a series of enzymes (caspases) that lead to cell death
78–81
. HighBcl-2 levels are associated with most types of humancancer and block the release of cytochromeC. Theyappear to be a contributor to both inherent and acquiredresistance to anticancer treatments. The
 BCL2
and
TP53
genes regulate
VEGF
-mediated angiogenesis
82
.
Curcumin and green tea extract inhibit
 BCL2
expression
83–85
.
Scutellaria baicalensis
contains thephenolic compounds baicalin, baicalein, wogenin,and oroxylin. These constituents inhibit
 BCL2
overexpression, plus
COX2
gene expression and
NF
-
κ 
B
activation
86,87
. Hibiscus protocatechuic acid is aphenolic compound isolated from the dried flower of 
 Hibiscus sabdariffa
L.; it inhibits Bcl-2 activity
88,89
.Other inhibitors of Bcl-2 include
EPA
from fish oil
90
,a lectin extract of 
Viscum album
(mistletoe)
91
, 6-gingerol
92
, grape seed extract
93
, echinocystic acid(a triterpene found in ginseng and other Asianherbs)
94,95
, parthenolide (a sesquiterpene lactonefound in feverfew)
96
, and beta-lapachone (a quinoneobtained from the bark of the lapacho tree)
97–99
.
2.7Targeting Coagulation Pathways Associatedwith Angiogenesis
In some clinical trials, anticoagulation drugs havebeen associated with a reduction in metastases
100–102
.
TABLE
 
I
Natural health products that inhibit cyclo-oxygenase-2activity
57
GingerAloe vera
EGCG
 /green teaResveratrolLiquoriceGarlicChinese skullcapBilberryGrape seed extract proanthocyanidinsPanax ginsengMilk thistleFish oils: omega-3 fatty acids (eicosapentaenoic acid,docosahexaenoic acid)Green-lipped musselAntioxidants (vitamins A, C, E; Se, Zn; carotenoids, flavonoids,coenzyme Q10,
-acetylcysteine, lipoic acid)BoswelliaBromelainCurcuminQuercetin
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