Project Report - Ganpat University Institutional Repository

1
Screening of natural compounds as Aromatase
inhibitors for treatment of Breast Cancer-An insilico
approach
Submitted By
Ankita R. Upadhyay
Ganpat University, Gujrat
Under the guidance of
Mr. Biplab Bhattacharjee
Manager –Operations
Institute Of Computational Biology
www.iocbindia.org
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ACKNOWLEDGEMENT
I, Ankita Upadhyay declare that this project entitled “Screening of
natural compounds as Aromatase inhibitors for treatment of Breast
Cancer-An insilico approach” has been prepared by me.
It is great pleasure for me to acknowledgement the assistance &
encouragement of a nu. of individuals in this effort. Mr. Biplab
Bhattacharjee, my project guide for offering valuable guidance&
encouragement during the entire project.
He has been a great help & his contribution in the successful
completion of this project.
I also thank to ShibhinSir & Jayadeepa Madam for their grateful
support.& thanks to my parents & my dear friend for their help & good
wishes.
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-: CONTENTS:-1. Introduction
1.1 Cancer
1.1.1 Treatment
1.2Breast Cancer
 1.2.1 Risk factor
 1.2.2 Signs &symptoms.
 1.2.3 Staging
 1.2.4 Test & Diagnosis
 1.2.5 Treatment
 1.3Drug discovery
 1.3.1 Methodologies
 1.4 Chemoprevention
 1.4.1 Breast Cancer Chemoprevention
 1.5 Nuclear Receptor Super family
 1.5.1Estrogen pathway
 1.6 Aromatase
 1.6.1 Action of Aromatase
 1.6.2 Role of Aromatase inhibitor
 1.6.3 Plant Sources for Aromatase Inhibitor
2. Review of Literature
3. Flowchart
 3.1Marerial & method
 3.1.1 Databases
o 3.1.1.1 Pub Med
o 3.1.1.2PDB
o 3.1.1.3Pubchem
 3.1.2 Software
o 3.1.2.1Marvin Sketch
o 3.1.2.2 Quantum3.0.0.
o 3.1.2.3 Argus lab
o 3.1.2.4 Hex 4.5
o 3.1.2.5ADME Tox
4. Result
 4.1 List of Aromatase inhibitor with Sources
 4.2 List of Aromatase inhibitor with compound
 4.3 Quantum, Argus Lab & Hex docking score
5. Discussion
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 5.1Table no.:4 Docking score:
 5.2Table no.:5. IC50 Result
 5.3Table no.:6 ADME-Tox properties
6. Conclusion
7. References
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LIST OF FIGURES:
FIGURE
NO.
FIGURE NAME
PAGE
NO.
1
2
Tumor cell
Endoscopy
7
13
3
Chemoprevention
32
4
5
Nuclear receptor structure
Structure of Estrogen (A), Estriol (B) and Estradiol (C)
38
38
6
Process of breast cancer
39
7
Actions of Aromatase
41
8
Plant Sources for Aromatase Inhibitor
44
9
Home page of Springer link
49
10
Home page of Pub med
50
11
12
13
14
15
16
The PDB home page of 3EQM
Enter the molecule name—Chrysin
Click the molecule, here
Compound Summary-Chrysin
Energy minimization for Tamoxifen in Marvin sketch
The conformers of Tamoxifen obtained after energy
minimization in Marvin sketch
54
55
56
56
57
58
17
Screen of Argus lab
58
18
Quantum homepage
59
19
Quantum homepage molecule
59
20
21
Quantum after loading the molecule
Sequence on (The sequence of the protein with the
water molecules is shown)
60
60
22
To remove the water molecule
61
6
23
24
After removing the water molecules
A create a object
61
62
25
26
27
28
29
30
A copy of the ligand is created
To rename the object
The object is renamed as ligand
The ligand being removed automatically after renaming
The ligand in the protein complex is selected
The hydrogen atoms are added and the protonation state
is set
62
63
63
64
64
65
31
The number of hydrogen atoms (26) and the electric state 65
(0e) is shown
32
The chain is selected by clicking on the chain
66
33
The grid center for docking is selected
66
34
35
The active site are highlighted
The protein showing the active site in red
67
67
36
37
Adding foreign ligand
Screening of foreign ligand
68
68
38
39
40
41
42
43
44
45
Argus lab-open the homepage.
Open the structure
Tree view of 3EQM
Hide unselected structure.
Center molecule in window.
View/center molecule in window.
Add hydrogen
Ctrl+c & ctrl+v the selected residue.
69
69
70
70
71
71
72
73
46
Right click on 603ASD & select “make a ligand group
from this residue” .now group have two ligands name “1
ASD” & “2ASD”.
73
47
Right click on “1 ASD” in the group folder & select
“modify group” option. In modify group dialog box,
type in a new name like ‘lig’
Right click on the group named “lig” & select the “make
a binding site group for this group
74
48
74
7
49
50
Selecting the Calculation/DOCK a Ligand
Settings dialog box looks like: select the ligand to dock
in the “Ligand “drop-box. Click on the “Calculate size”.
Make sure “ARGUS DOCK”. Than ok.
75
75
51
76
53
54
55
56
57
See Argus Lab first generate the scoring grids used
during the docking
Go to calculation in the calculations folder select the
Dock a Database
Click on Get Ligands & select the sdf compound.
See the result best Ligand phase-energy
Open the homepage & go to file open receptor...
Hex after loading the molecule
Open the homepage & go to file open Ligand...
58
59
60
61
62
63
64
Go to controls----- docking----- enter.
Docking control---activates ----hex progress
See the Hex progress
See the E total & rms
Save the result...
Save both---- enter the name ---ok
ADME Box
79
80
80
81
81
82
85
65
TOX Box
85
66
Graph
85
52
.
76
77
77
78
78
79
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LIST OF TABLES:
TABLE
NO
TABLE NAME
PAGE
NO.
1.
2.
List of Aromatase inhibitor with Sources
List of Aromatase inhibitor with compound properties:
86
128
3.
4.
5.
Quantum, Argus Lab & Hex Docking score
Docking score
IC50 Result:
136
141
143
6.
7.
ADME-Tox properties
Structure similarity with commercial drug
143
149
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1. Introduction
1.1 Cancer
Cancer is a class of diseases characterized by out-of-control cell growth.
There are over 100 different types of cancer, and each is classified by the
type of cell that is initially affected.
Fig_1 tumor cell
Cancer harms the body when damaged cells divide uncontrollably to
form lumps or masses of tissue called tumors (except in the case of
leukemia where cancer prohibits normal blood function by abnormal
cell division in the blood stream). Tumors can grow and interfere with
the digestive, nervous, and circulatory systems, and they can release
hormones that alter body function. Tumors that stay in one spot and
demonstrate limited growth are generally considered to be benign.
More dangerous, or malignant, tumors form when two things occur:
1. A cancerous cell manages to move throughout the body using the
blood or lymph systems, destroying healthy tissue in a process called
invasion
2. That cell manages to divide and grow, making new blood vessels to
feed itself in a process called angiogenesis.
When a tumor successfully spreads to other parts of the body and
grows, invading and destroying other healthy tissues, it is said to have
metastasized. This process itself is called metastasis, and the result is a
serious condition that is very difficult to treat.
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In 2007, cancer claimed the lives of about 7.6 million people in the
world. Physicians and researchers who specialize in the study,
diagnosis, treatment, and prevention of cancer are called oncologists.
How is cancer classified?
Cancers are classified by the type of cell that resembles the tumor and,
therefore, the tissue presumed to be the origin of the tumor. Examples
include:
Carcinoma: Malignant tumors derived from epithelial cells. This group
represents the most common cancers, including the common forms of
breast, prostate, lung and colon cancer.
Sarcoma: Malignant tumors derived from connective tissue, or
mesenchymal cells.
Lymphoma and leukemia: Malignancies derived from hematopoietic
(blood-forming) cells
Germ cell tumor: Tumors derived from totipotent cells. In adults it is
most oftenly found in the testicle and ovary; in fetuses, babies, and
young children most often found on the body midline, particularly at
the tip of the tailbone.
Blastic tumor: A tumor (usually malignant) which resembles an
immature or embryonic tissue. Many of these tumors are most common
in children.









Adeno- = gland
Chondro- = cartilage
Erythrocyte- = red blood cell
Hemangio- = blood vessels
Hepato- = liver
Lipo- = fat
Lympho- = white blood cell
Melano- = pigment cell
Myelo- = bone marrow
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




Myo- = muscle
Osteo- = bone
Uro- = bladder
Retino- = eye
Neuro- = brain
In the U.S. and other developed countries, cancer is presently
responsible for about 25% of all deaths. On a yearly basis, 0.5% of the
population is diagnosed with cancer. The statistics given in the table is
for adults in the United States, and may vary substantially in other
countries.
Cancer is the second leading cause of death in America, exceeded only
by heart disease. Every year, cancer is diagnosed in more than a million
people. The number of new cancer cases can be reduced substantially,
and many cancer deaths can be prevented. Adopting healthier
lifestyles—like avoiding tobacco use, increasing physical activity,
achieving optimal weight, improving nutrition, and avoiding sun
exposure—can significantly reduce a person's risk for cancer. Providing
cancer screening, information, and referral services available and
accessible to all individuals is also essential for reducing the high rates
of cancer and cancer deaths. Cancers that can be prevented or detected
earlier by screening account for about half of all cancer cases in the
United States.
What causes cancer?
Cancer is ultimately the result of cells that uncontrollably grow and do
not die. Normal cells in the body follow an orderly path of growth,
division, and death. Programmed cell death is called apoptosis, and
when this process breaks down, cancer begins to form. Unlike regular
cells, cancer cells do not experience programmatic death and instead
continue to grow and divide. This leads to a mass of abnormal cells that
grows out of control.
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Genes - the DNA type
Cells can experience uncontrolled growth if there are damages or
mutations to DNA, and therefore, damage to the genes involved in cell
division. Four key types of gene are responsible for the cell division
process: oncogenes tell cells when to divide, tumor suppressor genes tell
cells when not to divide, suicide genes control apoptosis and tell the cell
to kill itself if something goes wrong, and DNA-repair genes instruct a
cell to repair damaged DNA.
Cancer occurs when a cell's gene mutations make the cell unable to
correct DNA damage and unable to commit suicide. Similarly, cancer is
a result of mutations that inhibit oncogene and tumor suppressor gene
function, leading to uncontrollable cell growth.
Carcinogens
Carcinogens are a class of substances that are directly responsible for
damaging DNA, promoting or aiding cancer. Tobacco, asbestos, arsenic,
radiation such as gamma and x-rays, the sun, and compounds in car
exhaust fumes are all examples of carcinogens. When our bodies are
exposed to carcinogens, free radicals are formed that try to steal
electrons from other molecules in the body. Theses free radicals damage
cells and affect their ability to function normally.
What are the symptoms of cancer?
Cancer symptoms are quite varied and depend on where the cancer is
located, where it has spread, and how big the tumor is. Some cancers
can be felt or seen through the skin - a lump on the breast or testicle can
be an indicator of cancer in those locations. Skin cancer (melanoma) is
often noted by a change in a wart or mole on the skin. Some oral cancers
present white patches inside the mouth or white spots on the tongue.
Other cancers have symptoms that are less physically apparent. Some
brain tumors tend to present symptoms early in the disease as they
affect important cognitive functions. Pancreas cancers are usually too
small to cause symptoms until they cause pain by pushing against
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nearby nerves or interfere with liver function to cause a yellowing of the
skin and eyes called jaundice. Symptoms also can be created as a tumor
grows and pushes against organs and blood vessels. For example, colon
cancers lead to symptoms such as constipation, diarrhea, and changes in
stool size. Bladder or prostate cancers cause changes in bladder function
such as more frequent or infrequent urination.
As cancer cells use the body's energy and interfere with normal
hormone function, it is possible to present symptoms such as fever,
fatigue, excessive sweating, anemia, and unexplained weight loss.
However, these symptoms are common in several other maladies as
well. For example, coughing and hoarseness can point to lung or throat
cancer as well as several other conditions.
When cancer spreads, or metastasizes, additional symptoms can present
themselves in the newly affected area. Swollen or enlarged lymph nodes
are common and likely to be present early. If cancer spreads to the brain,
patients may experience vertigo, headaches, or seizures. Spreading to
the lungs may cause coughing and shortness of breath. In addition, the
liver may become enlarged and cause jaundice and bones can become
painful, brittle, and break easily. Symptoms of metastasis ultimately
depend on the location to which the cancer has spread.
How is cancer diagnosed and staged?
Early detection of cancer can greatly improve the odds of successful
treatment and survival. Physicians use information from symptoms and
several other procedures to diagnose cancer. Imaging techniques such as
X-rays, CT scans, MRI scans, PET scans, and ultrasound scans are used
regularly in order to detect where a tumor is located and what organs
may be affected by it. Doctors may also conduct an endoscopy, which is
a procedure that uses a thin tube with a camera and light at one end, to
look for abnormalities inside the body.
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Fig No.2 –Endoscopy
Extracting cancer cells and looking at them under a microscope is the
only absolute way to diagnose cancer. This procedure is called a biopsy.
Other types of molecular diagnostic tests are frequently employed as
well. Physicians will analyze your body's sugars, fats, proteins, and
DNA at the molecular level. For example, cancerous prostate cells
release a higher level of a chemical called PSA (prostate-specific antigen)
into the bloodstream that can be detected by a blood test. Molecular
diagnostics, biopsies, and imaging techniques are all used together to
diagnose cancer.
After a diagnosis is made, doctors find out how far the cancer has
spread and determine the stage of the cancer. The stage determines
which choices will be available for treatment and informs prognoses.
The most common cancer staging method is called the TNM system. T
(1-4) indicates the size and direct extent of the primary tumor, N (0-3)
indicates the degree to which the cancer has spread to nearby lymph
nodes, and M (0-1) indicates whether the cancer has metastasized to
other organs in the body. A small tumor that has not spread to lymph
nodes or distant organs may be staged as (T1, N0, M0), for example.
TNM descriptions then lead to a simpler categorization of stages, from 0
to 4, where lower numbers indicate that the cancer has spread less.
While most Stage 1 tumors are curable, most Stage 4 tumors are
inoperable or untreatable.
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1.1.1 Treatment
Cancer treatment depends on the type of cancer, the stage of the cancer
(how much it has spread), age, health status, and additional personal
characteristics. There is no single treatment for cancer, and patients
often receive a combination of therapies and palliative care. Treatments
usually fall into one of the following categories: surgery, radiation,
chemotherapy, immunotherapy, hormone therapy, or gene therapy.
Surgery
Surgery is the oldest known treatment for cancer. If a cancer has not
metastasized, it is possible to completely cure a patient by surgically
removing the cancer from the body. This is often seen in the removal of
the prostate or a breast or testicle. After the disease has spread,
however, it is nearly impossible to remove all of the cancer cells.
Surgery may also be instrumental in helping to control symptoms such
as bowel obstruction or spinal cord compression.
Radiation
Radiation treatment, also known as radiotherapy, destroys cancer by
focusing high-energy rays on the cancer cells. This causes damage to the
molecules that make up the cancer cells and leads them to commit
suicide. Radiotherapy utilizes high-energy gamma-rays that are emitted
from metals such as radium or high-energy x-rays that are created in a
special machine. Early radiation treatments caused severe side-effects
because the energy beams would damage normal, healthy tissue, but
technologies have improved so that beams can be more accurately
targeted. Radiotherapy is used as a standalone treatment to shrink a
tumor or destroy cancer cells (including those associated with leukemia
and lymphoma), and it is also used in combination with other cancer
treatments.
Chemotherapy
Chemotherapy utilizes chemicals that interfere with the cell division
process - damaging proteins or DNA - so that cancer cells will commit
suicide. These treatments target any rapidly dividing cells (not
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necessarily just cancer cells), but normal cells usually can recover from
any chemical-induced damage while cancer cells cannot. Chemotherapy
is generally used to treat cancer that has spread or metastasized because
the medicines travel throughout the entire body. It is a necessary
treatment for some forms of leukemia and lymphoma. Chemotherapy
treatment occurs in cycles so the body has time to heal between doses.
However, there are still common side effects such as hair loss, nausea,
fatigue, and vomiting. Combination therapies often include multiple
types of chemotherapy or chemotherapy combined with other treatment
options.
Immunotherapy
Immunotherapy aims to get the body's immune system to fight the
tumor. Local immunotherapy injects a treatment into an affected area,
for example, to cause inflammation that causes a tumor to shrink.
Systemic immunotherapy treats the whole body by administering an
agent such as the protein interferon alpha that can shrink tumors.
Immunotherapy can also be considered non-specific if it improves
cancer-fighting abilities by stimulating the entire immune system, and it
can be considered targeted if the treatment specifically tells the immune
system to destroy cancer cells. These therapies are relatively young, but
researchers have had success with treatments that introduce antibodies
to the body that inhibit the growth of breast cancer cells. Bone marrow
transplantation (hematopoetic stem cell transplantation) can also be
considered immunotherapy because the donor's immune cells will often
attack the tumor or cancer cells that are present in the host.
Hormone therapy
Several cancers have been linked to some types of hormones, most
notably breast and prostate cancer. Hormone therapy is designed to
alter hormone production in the body so that cancer cells stop growing
or are killed completely. Breast cancer hormone therapies often focus on
reducing estrogen levels (a common drug for this is tamoxifen) and
prostate cancer hormone therapies often focus on reducing testosterone
levels. In addition, some leukemia and lymphoma cases can be treated
with the hormone cortisone.
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Gene therapy
The goal of gene therapy is to replace damaged genes with ones that
work to address a root cause of cancer: damage to DNA. For example,
researchers are trying to replace the damaged gene that signals cells to
stop dividing (the p53 gene) with a copy of a working gene. Other genebased therapies focus on further damaging cancer cell DNA to the point
where the cell commits suicide. Gene therapy is a very young field and
has not yet resulted in any successful treatments.
How can cancer be prevented?
Cancers that are closely linked to certain behaviors are the easiest to
prevent. For example, choosing not to smoke tobacco or drink alcohol
significantly lower the risk of several types of cancer - most notably
lung, throat, mouth, and liver cancer. Even if you are a current tobacco
user, quitting can still greatly reduce your chances of getting cancer.
Skin cancer can be prevented by staying in the shade, protecting
yourself with a hat and shirt when in the sun, and using sunscreen. Diet
is also an important part of cancer prevention since what we eat has
been linked to the disease. Physicians recommend diets that are low in
fat and rich in fresh fruits and vegetables and whole grains.
Certain vaccinations have been associated with the prevention of some
cancers. For example, many women receive a vaccination for the human
papillomavirus because of the virus's relationship with cervical cancer.
Hepatitis B vaccines prevent the hepatitis B virus, which can cause liver
cancer.
Some cancer prevention is based on systematic screening in order to
detect small irregularities or tumors as early as possible even if there are
no clear symptoms present. Breast self-examination, mammograms,
testicular self-examination, and Pap smears are common screening
methods for various cancers.
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1.2Breast Cancer
What is Breast cancer?
Breast cancer is a malignant tumor that starts from cells of the breast. A
malignant tumor is a group of cancer cells that may grow into (invade)
surrounding tissues or spread (metastasize) to distant areas of the body.
The disease occurs almost entirely in women, but men can get it, too.
Cancer of the Breast can be defined as uncontrolled breast cell
growth.
. Cancer (medical term: malignant neoplasm) is a class of diseases in
which a group of cells display uncontrolled growth (division beyond the
normal limits), invasion (intrusion on and destruction of adjacent
tissues), and sometimes metastasis (spread to other locations in the body
via lymph or blood). These three malignant properties of cancers
differentiate them from benign tumors, which are self-limited, and do
not invade or metastasize. Most cancers form a tumor but some, like
leukemia, do not. The branch of medicine concerned with the study,
diagnosis, treatment, and prevention of cancer is oncology.
How does Breast cancer spread?
Breast cancer typically begins in the milk ducts of the breast. The cancer
can pierce through or invade the wall of the duct into the surrounding
breast tissue. There, the cancer can encounter microscopic blood vessels
and lymph channels. The cancer cells can break through the walls of
these vessels and travel outside the breast to other parts of the body
(including the bone, liver, lung, or brain). This can occur when the
cancer is barely able to be felt. This process is called metastasis.
When breast cancer spreads outside the breast, cancer cells are generally
found first in the lymph nodes under the arm. They have gotten there by
migrating from the primary tumor through lymph channels. Cancer
cells might also have spread to the bones, liver, lungs or brain. Cancer
that spreads to another part of the body is the same disease and has the
same name as the original cancer. When breast cancer spreads, it is
called metastatic breast cancer, even though it is found in another part
of the body.
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1.2.1 Risk factor
Breast cancer is actually the most common cancer of women. The
misconception people have is that they think it happens at a very young
age; actually the median age of breast cancer is 61, and so it's more
common in older women than younger women. It's a commonly quoted
statistic that 1 in 9 women will get breast cancer during their lifetime.
But it doesn't mean that every woman has a 1 in 9 chance of getting
breast cancer. Your risk is unique. It depends on lots of things.
A more useful way to think about the general risk of getting breast
cancer may be by age. The table below can give you some idea of your
chance of getting breast cancer over certain periods of your life.
Age
Risk of breast cancer
Up to age 25
1 in 15,000
Up to 30
1 in 1,900
Up to 40
1 in 200
Up to 50
Up to 60
Up to 70
Up to 80
1 in 50
1 in 23
1 in 15
1 in 11
Up to 85
Lifetime risk
1 in 10
1 in 9
Classification of Breast Cancer:
Breast cancers are described along four different classification
schemes, or groups, each based on different criteria and serving a
different purpose:
•Pathology - tumor based on its histological (microscopic anatomy)
appearance and other criteria. The most common pathologic types of
breast cancer are invasive ductal carcinoma, malignant cancer in the
breast's ducts, and invasive lobular carcinoma, malignant cancer in the
breast's lobules.
•Grade of tumor - A well-differentiated (low grade) tumor resembles
normal tissue. A poorly differentiated (high grade) tumor is composed
of disorganized cells and, therefore, does not look like normal tissue.
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Moderately differentiated (intermediate grade) tumors are somewhere
in between.
•Protein & gene expression status - Currently, all breast cancers should
be tested for expression, or detectable effect, of the estrogen receptor
(ER), progesterone receptor (PR) and HER2/neu proteins.
1.2.3 Staging
Cancer stage is based on the size of the tumor, whether the cancer is
invasive or non-invasive, whether lymph nodes are involved, and
whether the cancer has spread beyond the breast.
Stage 0
Stage 0 is used to describe non-invasive breast cancers, such as DCIS
and LCIS. In stage 0, there is no evidence of cancer cells or noncancerous abnormal cells breaking out of the part of the breast in which
they started, or of getting through to or invading neighboring normal
tissue.
Stage I
Stage I describes invasive breast cancer (cancer cells are breaking
through to or invading neighboring normal tissue) in which
The tumor measures up to 2 centimeters, AND
Stage II
No tumor can be found in the breast, but cancer cells are found in the
axillary lymph nodes (the lymph nodes under the arm), OR The tumor
measures 2 centimeters or less and has spread to the axillary lymph
nodes, The tumor is larger than 2 but no larger than 5 centimeters and
has spread to the axillary lymph nodes, OR the tumor is larger than 5
centimeters but has not spread to the axillary lymph nodes
Stage III
No tumor is found in the breast. Cancer is found in axillary lymph
nodes that are clumped together or sticking to other structures or cancer
may have spread to lymph nodes near the breastbone, OR the tumor is 5
centimeters or smaller and has spread to axillary lymph nodes that are
clumped together or sticking to other structures.
Stage IV
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"Metastatic at presentation" means that the breast cancer has spread
beyond the breast and nearby lymph nodes, even though this is the first
diagnosis of breast cancer. The reason for this is that the primary breast
cancer was not found when it was only inside the breast. Metastatic
cancer is considered stage IV
1.2.2 Signs &symptoms
There are often no symptoms associated with the early stage prostate
cancer. As the disease progresses and the tumour enlarge it may
compress and constrict the urethra which runs through the gland,
obstructing the flow of urine during urination.
In this situation the patient may notice a weak, interrupted stream of
urine that requires straining to urinate. On completion he may still feel
that the bladder is not empty.
However these symptoms are not specific to prostate cancer and are
most commonly found in benign non-cancerous enlargements of the
gland.
Blood in the semen may also be a sign of prostate cancer, although again
it is a common finding and not normally related to malignancy.
If tumors have spread to the bones then it may cause pain. The spine is
the most common site for this to occur. Although widespread use of
screening mammograms has increased the number of breast cancers
found before they cause any symptoms, some breast cancers are not
found by mammogram, either because the test was not done or because,
even under ideal conditions, mammograms do not find every breast
cancer. The most common sign of breast cancer is a new lump or mass.
A painless, hard mass that has irregular edges is more likely to be
cancerous, but breast cancers can be tender, soft, or rounded. For this
reason, it is important that any new breast mass or lump be checked by
a health care professional experienced in diagnosing breast diseases.
Other possible signs of breast cancer include:
* swelling of all or part of a breast (even if no distinct lump is felt)
* Skin irritation or dimpling
* Breast or nipple pain
* Nipple retraction (turning inward)
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* Redness, scaliness, or thickening of the nipple or breast skin
* A discharge other than breast milk
Sometimes a breast cancer can spread to underarm lymph nodes and
cause a lump or swelling there, even before the original tumor in the
breast tissue is large enough to be felt.
Why does Breast cancer occur?
Genetics
Researchers have isolated two genes that are common in a number of
cases where breast cancer runs in the family. It is the alteration (or
mutation) of these two genes that increases a person's risk of developing
breast cancer. Most of the time, it appears that the mutation is inherited.
For women, increased chances of developing breast cancer would be
due a mutation of either gene. In men, it is predominantly a mutation of
the BRCA2 gene (the other gene is called BRCA1).
Hormones can play a role in the chances of developing breast cancer,
more so in women than men, since it really relates to estrogen. When a
woman goes through menstruation, pregnancy or childbirth, her level of
estrogen fluctuates up and down. This hormone is basically telling cells
to divide. The more they divide, the more potential there is for this
division to create an abnormality in the new cells. Some of these
irregular cells could become cancerous. How this affects a woman's
chances of developing breast cancer is relates to the length of time she is
menstruating, the age she has her first child and the age she goes
through menopause. The risk increases when someone is very young
when she has her first period, older when she has first child and older
when she goes through menopause.
Lifestyle According to the American College of Physicians, people
(more so women than men) are more likely to develop breast cancer if
they eat a diet high in fat. Actually, it has been suggested that lowering
your calories from fat to below 30 percent for some and 20 percent for
others will greatly reduce your chances of breast cancer. There has also
been a suggestion that both obesity and alcohol abuse increase the
occurrence of breast cancer.
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Reduce the risk of Breast cancer
1. Pass on that last call for alcohol.
Studies have determined that women who drink alcoholic beverages
develop cancer at a higher rate. How much is too much? Based on
studies, ladies who consume 2 to 5 drinks daily have about 1½ times the
risk of women who don't consume alcohol.
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2. Quitters DO prosper - when it comes to smoking.
Although there has not been a direct link between smoking and breast
cancer, studies suggest that smoking at an early age can increase a
woman's risk. Not only can it be a risk for breast cancer, smoking is a
definite risk factor for lung cancer. Need help quitting? Click on over to
our Quit Smoking site, where you will find everything you need to
know to kick the habit.
3. Get physical.
Physical activity may reduce your risk of breast cancer. Studies by the
Women's Health Initiative found that women who walked briskly 1.25
to 2.5 hours per week reduced a woman's breast cancer risk by 18%.
Exercise doesn't always mean traditional gym exercises either. Check
out the Top 10 Ways to Prevent Cancer Through Exercise for fun ideas.
4. Be aware of your family breast cancer history.
Having a family or personal history of breast cancer may increase your
risk. If an immediate woman in your family has had breast cancer, it is
important to let your doctor know. Studies have shown that breast
cancer can be genetic. Genetic testing and counseling is available for
those concerned with their risk. Keep in mind, that just because your
mother or sister had breast cancer, it does not mean you will definitely
develop breast cancer.
24
5. Avoid hormone replacement therapy if possible.
Studies have shown a link between long time hormone replacement
therapy and breast cancer. This link suggests that combined HRT's
(estrogen and progesterone) raise the risk factor. Five years after
discontinuing HRT's the risk factor drops. HRT's also make
mammograms less effective. If you need to take hormone replacement
therapy, talk to your doctor about the risk and your personal condition.
6. Check your breasts every month.
Checking your breasts every month may not reduce your risk of
developing breast cancer, but it may help detect breast cancer early. The
earlier breast cancer is found, the less aggressive the treatment. Take a
look at "How To Perform a Self Breast Exam" to learn how to do an
exam, or to see if you are doing it correctly.
7. Try to keep a low fat diet.
A diet low in fat not only decreases the risk of obesity, it can reduce
your risk of breast cancer. We know that estrogen plays a major role in
the development of breast cancer. Fat tissue contains small amounts of
estrogen and may increase your risk. There have been conflicting studies
about fat intake and breast cancer risk, however all studies have
concluded that obesity plays a big part in breast cancer development.
8. Don't forget to get a mammogram - it's not a choice.
Like the breast self exam, a mammogram won't prevent the
development of breast cancer, but it can detect cancer. Sometimes it can
be difficult to feel a lump in the breast, and a mammogram is likely to
detect any lumps that cannot be felt.
9. Have children earlier in life, if possible
Having no children or having your first child in your mid-thirties or
later increases the risk.
10. Consider breastfeeding instead of formula feeding.
Researchers believe that the months without a period during pregnancy
and breast feeding may reduce a woman's risk of breast cancer. This
25
accompanies the data that suggests that early menopause lowers the risk
factor, as well.
Increase the risk of Breast cancer
Family history of breast cancer
Breast cancer risk is higher among women whose close blood relatives
have this disease.
Having one first-degree relative (mother, sister, or daughter) with breast
cancer approximately doubles a woman's risk. Having 2 first-degree
relatives increases her risk about 5-fold.
Although the exact risk is not known, women with a family history of
breast cancer in a father or brother also have an increased risk of breast
cancer. Altogether, about 20% to 30% of women with breast cancer have
a family member with this disease. This means that most (70% to 80%)
women who get breast cancer do not have a family history of this
disease.)
Personal history of breast cancer
A woman with cancer in one breast has a 3- to 4-fold increased risk of
developing a new cancer in the other breast or in another part of the
same breast. This is different from a recurrence (return) of the first
cancer.
Race and ethnicity
White women are slightly more likely to develop breast cancer than are
African-American women. African-American women are more likely to
die of this cancer. At least part of this seems to be because AfricanAmerican women tend to have more aggressive tumors, although why
this is the case is not known. Asian, Hispanic, and Native-American
women have a lower risk of developing and dying from breast cancer.
Dense breast tissue
Women with denser breast tissue (as seen on a mammogram) have more
glandular tissue and less fatty tissue, and have a higher risk of breast
cancer. Unfortunately, dense breast tissue can also make it harder for
doctors to spot problems on mammograms.
26
Certain benign breast conditions
Women diagnosed with certain benign breast conditions may have an
increased risk of breast cancer. Some of these conditions are more
closely linked to breast cancer risk than others. Doctors often divide
benign breast conditions into 3 general groups, depending on how they
affect this risk.
Proliferative lesions with atypia: In these conditions, there is excessive
growth of cells in the ducts or lobules of the breast tissue, and the cells
no longer appear normal. They have a stronger effect on breast cancer
risk, raising it 4 to 5 times higher than normal. They include:
* Atypical ductal hyperplasia (ADH)
* Atypical lobular hyperplasia (ALH)
Women with a family history of breast cancer and either hyperplasia or
atypical hyperplasia have an even higher risk of developing a breast
cancer.
For more information on these conditions, see the separate American
Cancer Society document, Non-cancerous Breast Conditions.
Women with lobular carcinoma in situ (LCIS) have a 7- to 11-fold
increased risk of developing cancer in either breast.
Menstrual periods
Women who have had more menstrual cycles because they started
menstruating at an early age (before age 12) and/or went through
menopause at a later age (after age 55) have a slightly higher risk of
breast cancer. This may be related to a higher lifetime exposure to the
hormones estrogen and progesterone.
Previous chest radiation
Women who, as children or young adults, had radiation therapy to the
chest area as treatment for another cancer (such as Hodgkin disease or
non-Hodgkin lymphoma) are at significantly increased risk for breast
cancer. This varies with the patient's age when they had radiation. If
chemotherapy was also given, it may have stopped ovarian hormone
27
production for some time, lowering the risk. The risk of developing
breast cancer from chest radiation is highest if the radiation was given
during adolescence, when the breasts were still developing. Radiation
treatment after age 40 does not seem to increase breast cancer risk.
Diethylstilbestrol exposure
From the 1940s through the 1960s some pregnant women were given the
drug diethylstilbestrol (DES) because it was thought to lower their
chances of miscarriage (losing the baby). These women have a slightly
increased risk of developing breast cancer. Women whose mothers took
DES during pregnancy may also have a slightly higher risk of breast
cancer. For more information on DES see the separate American Cancer
Society document, DES Exposure: Questions and Answers.
Lifestyle-related factors and breast cancer risk
Not having children, or having them later in life
Women who have had no children or who had their first child after age
30 have a slightly higher breast cancer risk. Having many pregnancies
and becoming pregnant at a young age reduce breast cancer risk.
Pregnancy reduces a woman's total number of lifetime menstrual cycles,
which may be the reason for this effect.
How is Breast cancer diagnosed?
Women with early breast cancer have no symptoms. This is why getting
the recommended screening tests before any symptoms develop is so
important. If something suspicious is found during a screening exam, or
if you have any of the symptoms of breast cancer described below, your
doctor will use one or more methods to find out if the disease is present.
If cancer is found, other tests will be done to determine the stage (extent)
of the cancer.
Drug target for Breast cancer:
Aromatase inhibitors stop the production of estrogen in postmenopausal women. Aromatase inhibitors work by blocking the
enzyme Aromatase, which turns another enzyme, androgen, into small
amounts of estrogen in the body. This means that less estrogen is
available to stimulate the growth of hormone-receptor-positive breast
cancer cells.
28
Aromatase inhibitors can't stop the ovaries from making estrogen, so
Aromatase inhibitors only work in post-menopausal women.
There are three Aromatase inhibitors:
•Arimidex (chemical name: anastrozole)
•Aromasin (chemical name: exemestane)
•Femara (chemical name: letrozole)
Each is a pill, usually taken once a day.
Benefits of Aromatase inhibitors
o A number of studies have compared Aromatase inhibitors with
tamoxifen to see which type of medicine was more effective in
treating early-stage, hormone-receptor-positive breast cancer in
post-menopausal women. Based on the results, most doctors
recommend that after initial treatment (surgery and possibly
chemotherapy and radiation therapy):
o An Aromatase inhibitor is the best hormonal therapy to start with.
When treating early-stage, hormone-receptor-positive breast
cancer, Aromatase inhibitors have more benefits and fewer serious
side effects than tamoxifen.
o Switching to an Aromatase inhibitor after taking tamoxifen for 2 to
3 years (for a total of 5 years of hormonal therapy) offers more
benefits than 5 years of tamoxifen.
o Taking an Aromatase inhibitor for 5 years after taking tamoxifen
for 5 years continues to reduce the risk of the cancer coming back,
compared to no treatment after tamoxifen.
Side effects of Aromatase inhibitors
o Aromatase inhibitors tend to cause fewer serious side effects than
tamoxifen, such as blood clots, stroke, and endometrial cancer. But
Aromatase inhibitors can cause more heart problems, more bone
loss (osteoporosis), and more broken bones than tamoxifen, at
least for the first few years of treatment. If you and your doctor are
considering an Aromatase inhibitor as part of your treatment plan,
you may want to ask your doctor about having a bone density test
to see if a bone strengthening medicine might be necessary while
you're taking the Aromatase inhibitor.
o The most common side effects of Aromatase inhibitors are joint
stiffness or joint pain.
29
o Joint pain from taking an Aromatase inhibitor can be troubling.
But a 2008 British study suggests that women who experienced
joint pain while taking hormonal therapy medicine were less
likely to have the breast cancer come back (recur). Knowing that
this side effect might indicate a reduced risk of the cancer coming
back may help some people stick with treatment despite the side
effects.
o If you're experiencing side effects from taking one Aromatase
inhibitor medicine, tell your doctor. You may be able to take a
different medicine. Arimidex and Femara have similar chemical
structures, while Aromasin has a different structure.
Aromatase inhibitors may cost more than tamoxifen
o Because Aromatase inhibitors are fairly new compared to
tamoxifen, much less is known about their long-term side effects
compared to tamoxifen. So far, no studies have directly compared
one Aromatase inhibitor to another.
o Aromatase inhibitors also cost more than tamoxifen, which is a
generic medicine. Aromatase inhibitors may cost hundreds of
dollars per month, compared to $40 to $100 per month for
tamoxifen. If you have financial concerns or don't have insurance
that will pay for an Aromatase inhibitor, you may be able to apply
for financial assistance. See the pages on Arimidex, Aromasin, and
Femara for more information.
1.4 Chemoprevention
Breast cancer is more frequent in nulliparous women, while its
incidence is significantly reduced by full-term pregnancy. The fact that
the protection conferred by pregnancy is observed in women from
different countries and ethnic groups, regardless of the endogenous
incidence of this malignancy, indicates that this protection does not
result from extrinsic factors specific to a particular environmental,
genetic, or socioeconomic setting, but rather from an intrinsic effect of
parity on the biology of the breast. Using an experimental system we
have shown that treatment of young virgin rats with human chorionic
gonadotropin (HCG), like full-term pregnancy, efficiently inhibits the
initiation and progression of chemically induced mammary carcinomas.
Treatment of young virgin rats with HCG induced a profuse lobular
30
development of the mammary gland, reduced the proliferative activity
of the mammary epithelium, and induced the synthesis of inhibin, a
secreted protein with tumor-suppressor activity. HCG treatment also
increased the expression of the programmed cell death (PCD) genes
testosterone repressed prostate message 2 (TRPM2), interleukin 1- converting enzyme (ICE), p53, c-myc, and bcl-XS, induced apoptosis,
and down regulated cyclins. PCD genes were activated through a p53dependent process, modulated by c-myc, and with partial dependence
on the bcl-2 family-related genes. The possibility that this hormonal
treatment activates known or new genes was tested by differential
display technique. We have identified a series of new genes, hormoneinduced-1 (HI-1) among them. The characterization of their functional
role will contribute to clarify the mechanisms through which hCG
inhibits the initiation and progression of mammary cancer. Of great
significance was the observation that PCD genes remained activated
even after lobular formations had regressed due to the cessation of
hormone administration. We postulate that this mechanism plays a
major role in the long-lasting protection exerted by hCG from
chemically induced carcinogenesis, and might be also involved in the
lifetime reduction in breast cancer risk induced in women by full-term
pregnancy. The implications of these observations are two-fold: on one
hand, they indicate that HCG, as pregnancy, may induce early genomic
changes that control the progression of the differentiation pathway, and
on the other, that these changes are permanently imprinted in the
genome, regulating the long-lasting refractoriness to carcinogenesis. The
permanence of these changes, in turn, makes them ideal surrogate
markers of HCG effect in the evaluation of this hormone as a breast
cancer preventive agent.
1.4.1 Breast Cancer Chemoprevention
Principles of Chemoprevention
Cancer chemoprevention is the inhibition, retardation or reversal of
carcinogenic processes by chemical means, and includes the treatment of
patients who have undergone successful treatment of a primary
malignancy but are at increased risk of a second. This latter concept is
designated “tertiary”chomoprevention. The prevention of
31
carcinogenesis at a premalignant stage is ‘primary’, and prevention at an
early phase of malignancy is ‘secondary’.
Several effective chemoprevention agents were first developed for
adjuvant chemotherapy; the prevention action of Tamoxifen, for
example, emerged when it was found to decrease breast tumours in the
contralateral breast after surgery. Such overlap between
chemotherapeutic and chemoprevention properties implies an
opportunity to conduct pilot studies of new chemopreventive agents in
patients who already have cancer, and this has been done with the
putative agents piroxicam,difluoromethylornithine(DFMO),and perillyl
alcohol.
Although the development of malignancy is the result of a complex
interaction between genetic and environmental influences, the multistep
model of carcinogenesis has provided incorporated. Existing paradigms
include clonal evolution and epigenetic changes, and these too offer
pointers to targets for chemoprevention agents. In colorectal neoplasia,
carcinogenesis has been characterized in exceptional detail.
32
Fig No.3 – Chemoprevention
1.5 Nuclear Receptor Super family
Nuclear receptors play many important roles in eukaryotic
development, differentiation, reproduction and metabolic homeostasis
(Ribeiro et al.,1995; Bain et al., 2006). Proteins of the nuclear receptor
super-family are single polypeptide chains with three major domains: a
variable amino-terminal domain, a highly conserved DNA-binding
domain (DBD), and a less conserved carboxyl-terminal ligand binding
33
domain (LBD). The superfamily is sub-divided into three classes. Class 1
is the steroid receptor family, and includes the progesterone receptor
(PR), the estrogen receptor (ER) (Figure 1), the glucocorticoid receptor
(GR), the androgen receptor (AR) and the mineralocorticoid receptor.
Class 2, or the thyroid/ retinoid family, includes the thyroid receptor
(TR), vitamin D receptor (VDR), the retinoic acid receptor (RAR) and the
peroxisome proliferator-activated receptor (PPAR). The third class of
nuclear receptors is known as the orphan receptor family. This class of
nuclear receptor comprises a set of proteins sharing significant sequence
homology to known nuclear receptors, but for which the ligands have
not yet been identified. Orphan nuclear receptor offer a unique system
for the discovery of novel signaling pathways that could provide new
drug targets for the treatment of a variety of human diseases.
Subfamily 1: Thyroid Hormone Receptor-like
•Group A: Thyroid hormone receptor (Thyroid hormone)
o
1: Thyroid hormone receptor-α (TRα; NR1A1, THRA)
o
2: Thyroid hormone receptor-β (TRβ; NR1A2, THRB)
•Group B: Retinoic acid receptor (Vitamin A and related compounds)
o
1: Retinoic acid receptor-α (RARα; NR1B1, RARA)
o
2: Retinoic acid receptor-β (RARβ; NR1B2, RARB)
o
3: Retinoic acid receptor-γ (RARγ; NR1B3, RARG)
•Group C: Peroxisome proliferator-activated receptor (fatty acids,
prostaglandins)
o
1: Peroxisome proliferator-activated receptor-α (PPARα; NR1C1,
PPARA)
o
2: Peroxisome proliferator-activated receptor-β/δ (PPARβ/δ;
NR1C2, PPARD)
o
3: Peroxisome proliferator-activated receptor-γ (PPARγ; NR1C3,
PPARG)
•Group D: Rev-ErbA (heme)
o
1: Rev-ErbAα (Rev-ErbAα; NR1D1)
o
2: Rev-ErbAβ (Rev-ErbAβ; NR1D2)
•Group F: RAR-related orphan receptor (cholesterol, ATRA)
34
o
o
o
1: RAR-related orphan receptor-α (RORα; NR1F1, RORA)
2: RAR-related orphan receptor-β (RORβ; NR1F2, RORB)
3: RAR-related orphan receptor-γ (RORγ; NR1F3, RORC)
•Group H: Liver X receptor-like (oxysterol)
o
1: Liver X receptor-α (LXRα; NR1H3)
o
2: Liver X receptor-β (LXRβ; NR1H2)
o
3: Farnesoid X receptor (FXR; NR1H4)
•Group I: Vitamin D receptor-like
o
1: Vitamin D receptor (VDR; NR1I1, VDR) (vitamin D)
o
2: Pregnane X receptor (PXR; NR1I2) (xenobiotics)
o
3: Constitutive androstane receptor (CAR; NR1I3) (androstane)
Subfamily 2: Retinoid X Receptor-like
•Group A: Hepatocyte nuclear factor-4 (HNF4) (fatty acids)
o
1: Hepatocyte nuclear factor-4-α (HNF4α; NR2A1, HNF4A)
o
2: Hepatocyte nuclear factor-4-γ (HNF4γ; NR2A2, HNF4G)
•Group B: Retinoid X receptor (RXRα) (retinoids)
o
1: Retinoid X receptor-α (RXRα; NR2B1, RXRA)
o
2: Retinoid X receptor-β (RXRβ; NR2B2, RXRB)
o
3: Retinoid X receptor-γ (RXRγ; NR2B3, RXRG)
•Group C: Testicular receptor
o
1: Testicular receptor 2 (TR2; NR2C1)
o
2: Testicular receptor 4 (TR4; NR2C2)
•Group E: TLX/PNR
o
1: Human homologue of the Drosophila tailless gene (TLX;
NR2E1)
o
3: Photoreceptor cell-specific nuclear receptor (PNR; NR2E3)
•Group F: COUP/EAR
o
1: Chicken ovalbumin upstream promoter-transcription factor I
(COUP-TFI; NR2F1)
o
2: Chicken ovalbumin upstream promoter-transcription factor II
(COUP-TFII; NR2F2)
35
o
6: V-erbA-related gene|V-erbA-related (EAR-2; NR2F6)
Subfamily 3: Estrogen Receptor-like
•Group A: Estrogen receptor (Sex hormones: Estrogen)
o
1: Estrogen receptor-α (ERα; NR3A1, ESR1)
o
2: Estrogen receptor-β (ERβ; NR3A2, ESR2)
•Group B: Estrogen related receptor
o
1: Estrogen-related receptor-α (ERRα; NR3B1, ESRRA)
o
2: Estrogen-related receptor-β (ERRβ; NR3B2, ESRRB)
o
3: Estrogen-related receptor-γ (ERRγ; NR3B3, ESRRG)
•Group C: 3-Ketosteroid receptors
o
1: Glucocorticoid receptor (GR; NR3C1) (Cortisol)
o
2: Mineralocorticoid receptor (MR; NR3C2) (Aldosterone)
o
3: Progesterone receptor (PR; NR3C3, PGR) (Sex hormones:
Progesterone)
o
4: Androgen receptor (AR; NR3C4, AR) (Sex hormones:
Testosterone)
Subfamily 4: Nerve Growth Factor IB-like
•Group A: NGFIB/NURR1/NOR1
o
1: Nerve Growth factor IB (NGFIB; NR4A1)
o
2: Nuclear receptor related 1 (NURR1; NR4A2)
o
3: Neuron-derived orphan receptor 1 (NOR1; NR4A3)
Subfamily 5: Steroidogenic Factor-like
•Group A: SF1/LRH1
o
1: Steroidogenic factor 1 (SF1; NR5A1) (phospholipids)
o
2: Liver receptor homolog-1 (LRH-1; NR5A2)
Subfamily 6: Germ Cell Nuclear Factor-like
•Group A: GCNF
o
1: Germ cell nuclear factor (GCNF; NR6A1)
Subfamily 0: Miscellaneous
36
•Group B: DAX/SHP
o
1: Dosage-sensitive sex reversal, adrenal hypoplasia critical region,
on chromosome X, gene 1 (DAX1, NR0B1)
o
2: Small heterodimer partner (SHP; NR0B2)
•Group C: Nuclear receptors with two DNA binding domains (2DBDNR)]
Nuclear receptors and mechanisms of signaling
All nuclear receptors modulate gene transcription, although amongst
the three classes there are differences in the mechanisms through which
this is achieved (Ribeiro, 1995; Aranda and Pascual, 2001; Bain et al.,
2006). Ligands for nuclear receptors circulate in the body bound to
plasma proteins. Following dissociation from these proteins the ligands
enter cells and bind to their receptors. Steroids and vitamin D probably
enter cells through passive diffusion, whereas thyroid hormone and
retinoic acid might gain cellular entrance via specific transport
processes.
Steroid receptors are bound to Hsps, such as Hsp90 and Hsp70, in the
cytoplasm. Upon binding ligand, the free receptors then translocate to
the nucleus and bind as homodimers to imperfect palindromic response
elements at upstream promoter sites. DNA binding is coupled to the
recruitment of transcriptional co-activators such as the p160 family (Xu
and Li, 2003). Nuclear receptors can in some instances repress gene
expression in a ligand-dependent manner, and in some cases promote
gene transcription.
The class 2 nuclear receptors typically function as heterodimers. TR,
VDR, RAR and PPAR associate with the retinoid X receptor (RXR) and
bind as a dimeric complex to direct repeat response elements (Ribeiro,
1995; Aranda and Pascual, 2001; Eckey et al., 2003; Bain et al., 2006). The
heterodimers are bound to their response element regardless of whether
ligands are present and in the absence of heat shock proteins. Gene
37
activation is suppressed by co-repressors such as silencing mediator for
retinoic acid and thyroid hormone receptors (SMRT) and nuclear corepressor (NCoR). Co-repressors are displaced by ligand binding
allowing transcriptional activation to take place (Collingwood et al.,
1999).
Nuclear receptor structure
A typical nuclear receptor consists of a variable NH2 terminal region
(the A/B domain) and a highly conserved DNA-binding domain (DBD
or C domain) (Tata, 2002; Robinson-Rechavi et al., 2003; Bain et al.,
2006). The DBD contains a P-box, which is a short motif responsible for
DNA-binding specificity and is involved in dimerization of nuclear
receptors including the formation of both heterodimers and
homodimers. The elucidiation of the 3D structure of the DBD reveals the
presence of two zinc fingers.
A linker region known as domain D is situated between the DBD and
the ligand binding domain. This region functions as a flexible hinge and
contains the nuclear localization signal. Phosphorylation of the hinge
region is coupled with increased transcriptional activation. The ligand
binding domain (LBD or E domain) is responsible for the binding of
cognate ligand or hormone. This domain also contains a ligandregulated transcriptional activation function (AF-2) necessary for
recruiting transcriptional co-activators, which interact with chromatin
remodeling proteins and the general transcriptional activation
machinery. Most nuclear receptors contain an amino acid sequence Nterminal to the DBD in the variable A/B domain, which contains a
transcriptional activation function known as AF-1. In contrast to the
moderately conserved AF-2, the AF-1 shows weak conservation across
the nuclear receptor super-family and may mediate differential
promoter regulation in vivo. The AF-1 sequence functions as a ligandindependent transcriptional activator, but can also functionally
synergize with AF-2. In addition to the domains described above some
receptors contain a carboxyl-terminal region (F domain) of unknown
function.
38
Proteins of the nuclear receptor super-family are single polypeptide
chains consisting of three major domains: a variable amino-terminal
domain, a highly conserved DNA-binding domain (DBD), and a less
conserved carboxyl-terminal ligand binding domain (LBD). AF:
activation function (domain 1 and 2). H: hinge. Mutations in the
nuclear hormone receptors lead to a variety of inherited disorders.
Mutations in X-linked ARs result in testicular feminization
syndrome with androgen unresponsiveness or hypo-responsiveness.
Glucorticoid resistance and hereditary vitamin D resistant rickets are
rare autosomal recessive disorders linked to mutations in the LBD or
the DBD of the GR or the VDR respectively (Ribeiro et al., 1995).
Fig No.4– Nuclear receptor structure
1.5.1Estrogen pathway
(A)
(B)
(C),.
Figure 5: Structure of Estrogen (A), Estriol (B) and Estradiol (C)
39
Estrogen plays a critical role in the development of breast cancer,
because it can stimulate the growth of breast tissues. If the breast
contains abnormal cells, they may also be stimulated by estrogen to
proliferate and become a tumor.
The estrogen receptor (ER) is a member of the nuclear hormone family
of intracellular receptors which is activated by the hormone 17βestradiol.[24] The main function of the estrogen receptor is as a DNA
binding transcription factor which regulates gene expression.
Fig No.6– process of breast cancer
40
1.6 Aromatase
Aromatase is an enzyme that is responsible for a key step in the
biosynthesis of estrogens. Because estrogens also promote certain
cancers and other diseases, aromatase inhibitors are frequently used to
treat those diseases.Steroids are composed of four fused rings (labeled
A, B, C, and D in the figure below). Aromatase transforms the left-hand
ring (the A-ring) of steroids to an aromatic state (hence the name)
through oxidation and subsequent elimination of a methyl
group.Aromatase converts testosterone to estradiol
Aromatase is a member of the cytochrome P450 superfamily (EC
1.14.14.1), whose function is to aromatize androgens, producing
estrogens. As such, it is an important factor in sexual development.
Cellular and tissue location
The enzyme is located in the endoplasmic reticulum of the cell and its
activity is regulated by tissue specific promoters that are in turn
controlled by hormones, cytokines, and other factors. The principal
action of the enzyme transforms androstenedione to estrone and
testosterone to estradiol. The aromatase enzyme can be found in many
tissues including gonads, brain, adipose tissue, placenta, blood vessels,
skin, bone, endometrium as well as in tissue of endometriosis, uterine
fibroids, breast cancer, and endometrial cancer.
Activity
Factors known to increase aromatase activity include age, obesity,
insulin, gonadotropins, and alcohol. Aromatase activity is decreased by
prolactin, anti-müllerian hormone, and smoking. Aromatase activity
appears to be enhanced in certain estrogen-dependent local tissue next
to breast tissue, endometrial cancer, endometriosis, and uterine fibroids.
Disorders
Aromatase excess syndrome
A number of investigators have reported on a rather rare syndrome of
excess aromatase activity. In boys it can lead to gynecomastia and in
girls to precocious puberty and gigantomastia. In both sexes, early
epiphyseal closure leads to shortness.
Aromatase deficiency syndrome
 This syndrome is due to a mutation of gene CYP19 and inherited
in an autosomal recessive way. Accumulations of androgens
during pregnancy may lead to virilization of a female at birth
41
(males are not affected). Females will have primary amenorrhea.
Individuals of both sexes will be tall as lack of estrogen does not
bring the epiphyseal lines to closure.
1.6.1 Action of Aromatase
Fig No.7 – Actions of Aromatase
Aromatase deficiency:
This syndrome is due to a mutation of gene CYP19 and inherited in an
autosomal recessive way. Accumulations of androgens during
pregnancy may lead to virilization of a female at birth (males are not
affected). Females will have primary amenorrhea. Individuals of both
sexes will be tall as lack of estrogen does not bring the epiphyseal lines
to closure.
42
1.6.2 Role of Aromatase inhibitor
Aromatase inhibitors stop the production of estrogen in postmenopausal women. Aromatase inhibitors work by blocking the
enzyme aromatase, which turns another enzyme, androgen, into small
amounts of estrogen in the body. This means that less estrogen is
available to stimulate the growth of hormone-receptor-positive breast
cancer cells.
There are three aromatase inhibitors:
* Arimidex (chemical name: anastrozole)
* Aromasin (chemical name: exemestane)
* Femara (chemical name: letrozole)
Each is a pill, usually taken once a day.
Benefits of aromatase inhibitors
A number of studies have compared aromatase inhibitors with
Tamoxifen to see which type of medicine was more effective in treating
early-stage, hormone-receptor-positive breast cancer in postmenopausal women. Based on the results, most doctors recommend that
after initial treatment (surgery and possibly chemotherapy and radiation
therapy):
* An aromatase inhibitor is the best hormonal therapy to start with.
When treating early-stage, hormone-receptor-positive breast cancer,
aromatase inhibitors have more benefits and fewer serious side effects
than Tamoxifen.
* switching to an aromatase inhibitor after taking Tamoxifen for 2 to 3
years (for a total of 5 years of hormonal therapy) offers more benefits
than 5 years of Tamoxifen.
* taking an aromatase inhibitor for 5 years after taking Tamoxifen for 5
years continues to reduce the risk of the cancer coming back, compared
to no treatment after Tamoxifen.
Side effects of aromatase inhibitors
Aromatase inhibitors tend to cause fewer serious side effects than
Tamoxifen, such as blood clots, stroke, and endometrial cancer. But
aromatase inhibitors can cause more heart problems, more bone loss
(osteoporosis), and more broken bones than Tamoxifen, at least for the
first few years of treatment. If you and your doctor are considering an
aromatase inhibitor as part of your treatment plan, you may want to ask
your doctor about having a bone density test to see if a bone
43
strengthening medicine might be necessary while you're taking the
aromatase inhibitor.
The most common side effects of aromatase inhibitors are joint stiffness
or joint pain.
Joint pain from taking an aromatase inhibitor can be troubling. But a
2008 British study suggests that women who experienced joint pain
while taking hormonal therapy medicine were less likely to have the
breast cancer come back (recur). Knowing that this side effect might
indicate a reduced risk of the cancer coming back may help some people
stick with treatment despite the side effects.
If you're experiencing side effects from taking one aromatase inhibitor
medicine, tell your doctor. You may be able to take a different medicine.
Arimidex and Femara have similar chemical structures, while Aromasin
has a different structure.
Aromatase inhibitors may cost more than Tamoxifen because aromatase
inhibitors are fairly new compared to Tamoxifen, much less is known
about their long-term side effects compared to Tamoxifen. So far, no
studies have directly compared one aromatase inhibitor to another.
Aromatase inhibitors also cost more than Tamoxifen, which is a generic
medicine. Aromatase inhibitors may cost hundreds of dollars per
month, compared to $40 to $100 per month for Tamoxifen. If you have
financial concerns or don't have insurance that will pay for an aromatase
inhibitor, you may be able to apply for financial assistance. See the
pages on Arimidex, Aromasin, and Femara for more information.
44
1.6.3 Plant Sources for Aromatase Inhibitor
Lemon
Dark chocolate
Cauliflower
Green tea
Citrus
Astragalus
Aloe Vera
Soybean oil
Peanuts
Broccoli
Curcuma domestica
Agaricus bisporus
Fig No.8:Plant Sources for Aromatase Inhibitor
45
3.Review of Literature
1. Chrysin: Chrysin, also known as flavone X, inhibits aromatase, an
enzyme responsible for breaking down testosterone into estrogen,
thereby helping to maintain higher testosterone levels. chrysin can be
taken alone or used along with other testosterone-boosting agents, such
as androstenedione and DHEA. There are few human studies on
chrysin; however, European Olympic athletes who took 1 to 3 grams of
chyrsin reportedly experienced a 30 percent increase in testosterone
2. Phytoestrogen: Phytoestrogens are a group of plant-derived
substances that are structurally or functionally similar to
estradiol.1,2 Interest in phytoestrogens, particularly soy, has been fueled
by epidemiologic studies that have suggested low incidence of breast
cancer in countries with high soy intake, and this has been followed
by in vitro and in vivo animal research suggesting a potential role for
phytoestrogens in preventing breast cancer development.Dietary
changes present one of the few socially acceptable modifiable risk
factors for breast cancer, the second leading cause of cancer deaths in
women.Hence, even a modest protective role of phytoestrogens could
have important implications for publichealth.
3. Quercetin: Quercetin is the main reason an apple a day keeps the
doctor away. It is a major antioxidant with important anti-aging
benefits. It fights inflammation and reduces the cellular damage
inflammation causes. By fighting inflammation, it also helps decrease
swelling and pain, and keeps the circulatory system healthy.. It is an
anti-viral, and an immune system supporter and liver protector.
Research has suggested that quercetin has other anti-cancer benefits
aside from inhibiting aromatase in breasts and prostates. Cabbage,
onions and garlic are other good sources of this powerful flavonoid.
3.http://www.tmuscle.com/free_online_article/sports_body_training_performance_body
building_supplements/naturally_occurring_aromatase_inhibitors
4 Kadohama, N., K. Shintani, and Y. Osawa, Tobacco alkaloid derivatives as
inhibitors of breast cancer aromatase. Cancer Lett, 1993. 75(3): p. 175-182.
Osawa, Y., et al., Aromatase inhibitors in cigarette smoke, tobacco leaves and
other plants. J Enzyme Inhib, 1990. 4(2): p. 187-200
5Preventive effect of the flavonoid, quercetin, on hepatic cancer in rats via
oxidant/antioxidant activity: molecular and histological evidences.
http://www.raysahelian.com/aromatase.html
46
4. Naringenin: Naringenin, is an antioxidant, free radical scavenger,
anti-inflammatory, and immune system modulator. It has been shown
to promote proper metabolism of carbohydrates. It was shown to reduce
hepatitis C virus production by infected liver cells in cell culture and to
inhibit the secretion of very low density lipoprotein by cells. As a cancer
fighter, it reduces oxidative damage to DNA. Naringenin is found in all
citrus and may be the reason that diets high in citrus are negatively
correlated with heart disease. However, naringenin should not be
obtained from grapefruit or grapefruit juice, which has an inhibitory
effect on the human cytochrome P450 isofrom, another enzyme in the
same complex as the aromatase enzyme. This enzyme is involved in
breaking down and metabolizing sex hormones and preventing their
excess accumulation in the body, so inhibiting it is not a good idea.
5. apigenin: Apigenin is a non-mutigenic flavonoid that has significant
chemoprotective action against UV radiation. Research has shown
apigenin reduces oxidative damage of DNA, inhibits the growth and
induces differentiation in human leukemia cells, inhibits cancer cell
transduction, and induces appropriate cell death. Like quercetin,
apigenin acts as an anti-inflammatory and as an antispasmodic.
Apigenin is found in good supply in celery, parsley, artichokes, basil,
and chamomile
6. Genistein: We conducted a range-finding experiment to determine
the appropriate dosage of androstenedione and Letrozole that will be
used in the silastic implants. Next we evaluated effect of dietary
genistein on the growth of estrogen-dependent breast tumors using an
intratumoral aromatase expressing postmenopausal breast cancer (MCF7Ca) model.. We identified an appropriate dosage of Letrozole and
found that dietary genistein will stimulate the growth of MCF-7Ca
tumors in the presences of Letrozole. Therefore, caution is necessary for
postmenopausal women with estrogen-dependent breast cancer
consuming dietary genistein.
6. http://www.drugbank.ca/drugs/DB03467
http://www.ncbi.nlm.nih.gov/sites/entrez
http://www.raysahelian.com/aromatase.html
8. http://en.wikipedia.org/wiki/Apigenin
http://www.raysahelian.com/aromatase.html
9. http://www.drugbank.ca/drugs/DB01645
http://www.raysahelian.com/aromatase.html
47
7. Astragalus: The success of the Als on breast cancer and their potential
effect on other estrogen-responsive diseases have led our team of
researchers to explore possible alternative Als in herbs. What we
discovered was that the herbal combination of Curcuma, Cyperus, and
Astragalus exhibits aromatase inhibition activity in rats, as presented in
the following sections. This combination of herbs will be referred to as
Myomin in this article. The succeeding in vitro, in vivo, and clinical
studies will demonstrate Myomin's mechanism, functions, and
applications.
8. Emoidin: Alnus glutinosa (buds): Contains Emodin, as antiaggregant, anti-inflammatory, antimutagenic, antiseptic, antitumor
(yeast, leukemia, oral, prostate). Emodin treatment has shown to
repress androgen-dependent transactivation of AR by inhibiting AR
nuclear translocation. Emodin decreases the association of AR and heat
shock protein 90 and increases the association of AR and MDM2, which
in turn induces AR degradation through proteasome-mediated pathway
in a ligand-independent manner. (See more about androgens under
Silver Birch.) Emodin is capable of inhibiting cellular proliferation,
inducing apoptosis, and preventing metastasis.
9. Ellagic acid: Estrogen receptor alpha (ERalpha)positive breast cancers that co-express transcription factors GATA-3
and FOXA1 have a favorable prognosis. These transcription factors form
an autoregulatory hormonal network that influences estrogen
responsiveness and sensitivity to hormonal therapy.[...]In this study, we
report that insulin increases the expression of Tbet in breast cancer cells, which correlates with reduced expression
of GATA-3, FOXA1, and the ERalpha:[...]-positive cells with
intrinsic tamoxifen resistance as well as MCF-7 cells with
acquired tamoxifen and fulvestrant resistance expressed elevated levels
of T-bet and/or reduced levels of FOXA1 and GATA-3.
11. http://www.drugbank.ca/drugs/DB00668
http://findarticles.com/p/articles/mi_7396/is_317/ai_n45060728/
http://www.herbalremedies.com/estrogenex.html
13. http://www.freepatentsonline.com/y2004/0156926.html
46. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=529&Title=Ellagic%20Acid
48
3. Flowchart
Collection of Molecules via Literature Study
Retrieval of Structures from PubChem
Energy minimization using Marvin Sketch
Retrieval of Receptor from PDB
Docking in Quantum, Hex & Argus Lab
Find ADME TOX Properties
49
Collection of Molecules via Literature Study
 Collection information regarding chemoprevention of Breast
Cancer.
 Searching for chemo preventive molecules using databases,
journals & articles.
 Pub Med
 Springer link
 Biomed central
 Science Direct
 Nature
 Filtering the search by selecting only naturally derived aromatase
inhibitor which are not commercialized.
Figure: 9: Home page of Springer link
50
Figure: 10: Home page of Pub med
51
3.1Marerial & method
DATABASES: PUBMED
 PDB
 PUBCHEM
SOFTWARES: MARVIN SKETCH
 ARGUS LAB
 QUANTUM
 HEX
 ADME TOX
52
3.1.1 Databases
3.1.1.1 Pub Med
http://www.ncbi.nlm.nih.gov/pubmed
Pub Med is a free resource that is developed and maintained by the
National Center for Biotechnology Information (NCBI), at the U.S.
National Library of Medicine (NLM), located at the National Institutes
of Health (NIH).
PubMed Coverage
PubMed provides access to bibliographic information that includes
MEDLINE, as well as: The out-of-scope citations (e.g., articles on plate
tectonics or astrophysics) from certain MEDLINE journals, primarily
general science and chemistry journals, for which the life sciences
articles are indexed for MEDLINE. Citations that precede the date that a
journal was selected for MEDLINE indexing. Some additional life
science journals that submit full text to PubMedCentral and receive a
qualitative review by NLM.
PubMed Journal Information
The Journals Database can be searched by subject or by using the
journal title, the Title Abbreviation, the NLM ID (NLM's unique journal
identifier), the International Organization for Standardization (ISO)
abbreviation, and the print and electronic International Standard Serial
Numbers (pISSNs and eISSNs). The database includes journals in all
Entrez databases (e.g., PubMed, Nucleotide, and Protein).
PubMed Citation Matchers
Use the Single Citation Matcher for finding the citation for a
particular article using title words or citation information, or to find an
entire volume or issue of a journal, or to generate a bibliography by a
first author.
The Batch Citation Matcher allows users to match their own list of
citations to PubMed citations, using bibliographic information such as
journal, volume, issue, page number, and year. The Citation Matcher
returns the corresponding PMID. This number can then be used to
easily link to PubMed
53
3.1.1.2PDB
http://www.rcsb.org/pdb
The Protein Data Bank (PDB) archive is repository for 3D
structural data of proteins and nucleic acids. The PDB archive contains
information about experimentally-determined structures of proteins,
nucleic acids, and complex assemblies. As a member of the wwPDB, the
RCSB PDB curates and annotates PDB data according to agreed upon
standards. The RCSB PDB also provides a variety of tools and resources.
Users can perform simple and advanced searches based on annotations
relating to sequence, structure and function. These molecules are
visualized, downloaded, and analyzed by users who range from
students to specialized scientists.
The Worldwide PDB - wwPDB consist of organization that act as
deposition, data processing and distribution centers for PDB data. The
RCSB PDB is a member of the wwPDB, a collaborative effort with PDBe
(UK), PDBj (Japan), and BMRB (USA) to ensure the PDB archive is
global and uniform. The protein data bank was used to retrieve the
estrogen receptor protein (3ert) for the docking studies on breast cancer..
54
Figure 11: The PDB home page of 3EQM
55
3.1.1.3Pubchem
http://pubchem.ncbi.nlm.nih.gov/
PubChem is a database of chemical molecules. The system is
maintained by the National Center for Biotechnology Information
(NCBI), a component of the National Library of Medicine, which is part
of the United States National Institutes of Health (NIH). PubChem
contains substance descriptions and small molecules with fewer than
1000 atoms and 1000 bonds.
PubChem provides information on the biological activities of
small molecules. It is a component of NIH's Molecular Libraries
Roadmap Initiative.PubChem includes substance information,
compound structures, and BioActivity data in three primary databases,
Pcsubstance, Pccompound, and PCBioAssay, respectively. The
PubChem substance database contains chemical structures, synonyms,
registration IDs, description, related urls, database cross-reference links
to PubMed, protein 3D structures, and biological screening results. If the
contents of a chemical sample are known, the description includes links
to PubChem Compound.
Figure: 12- Enter the molecule name—Chrysin.
56
Figure: 13 Click the molecule,here
Figure: 14- Compound Summary-Chrysin
57
3.1.2 Software
3.1.2.1Marvin Sketch
Marvin sketch is a customizable GUI with a brand new design.
The features of Marvin sketch are Configuration choices (including
ISIS/Draw and ChemDraw like menu and icon arrangements),Chain
drawing, displaying the last carbon number, Enhanced Query, S-group
and R-group drawing features, Global IME support in Textboxes (allows
entering East Asian characters) etc. The Lipinski properties were also
calculated using Marvin Sketch.
Figure:15 Energy minimization for Tamoxifen in Marvin sketch
58
Figure: 16 The conformers of Tamoxifen obtained after energy
minimization in Marvin sketch
3.2.2 Argus lab:
Figure: 17 Screen of Argus lab
59
3.1.2.2 Quantum3.3.0.
Quantum is a software package of drug discovery and
computational chemistry tools. Docking of each natural alternative
against the target estrogen receptor alpha and the docking scores were
done and saved
The protocol to work with quantum is as follow:-
Figure: 18: Quantum homepage
Step 1: To load the molecule
Fileopen moleculeselect the appropriate molecule from the
folder.
60
Figure 20: Quantum after loading the molecule
Step 2: Sequence on
Click on the “sequence off” tab”sequence on”
Figure 21: Sequence on (The sequence of the protein with the water
molecules is shown)
Step 3: To remove water molecules
Actionsremove water
61
Figure 22: To remove the water molecule
Figure 23: After removing the water molecules
Step 4: To create a copy of the ligand
62
Right click on the ligandcreate object
Figure 24: A create a object
Figure 25: A copy of the ligand is created
63
Step 5: To rename the object created
Actionsrename objectrename as “ligand”
Figure 26: To rename the object
Figure 27: The object is renamed as ligand
64
Step 6: To prepare the ligand for docking
Small molecules/ionsnewselectselect the ligandnext
(Hydrogen atoms are added and the protonation state is set)next (The
number of hydrogen atoms and electric charge is obtained)
Figure 28: The ligand being removed automatically after renaming
Figure 29: The ligand in the protein complex is selected
65
Figure 30: The hydrogen atoms are added and the protonation state is
set
Figure 31: The number of hydrogen atoms (26) and the electric state
(0e) is shown
66
Step 7: To select the chain
Biomacromoleculesnewselectdocking /screeningselect the
chain(by clicking on the any part of the protein)
Figure 32: The chain is selected by clicking on the chain
Step 8: To select the grid center
Click on the protein to select the grid centeradjust the position of the
grid boxnext
Figure 33: The grid center for docking is selected
67
Figure 34: The active site are highlighted
Step 9: Docking
Projectligand dockingselect the protein and the ligandset for
10 conformersstart dockingshow results
Figure 35: The protein showing the active site in red
Step 10: To add foreign ligands
68
Small moleculesnewchoose
molecules from the appropriate folder
directoryselect
the
set
of
Figure 36: Adding foreign ligands
Step 11:
Projectlibrary screeningselect the protein and the ligandsset
for 10 conformersstart screening
Figure 37: Screening of foreign ligand
69
3.2.3ARGUS LAB
Argus Lab is a molecular modeling program that runs on Window,
Vista.ArgusLAb consist of a user interface that supports OpenGL
graphics display of molecule structures and runs quantum mechanical
calculation using the Argus compute server. The Argus compute server
is constructed using the Microsoft Component Object Model (COM).
Argus Lab Features: -- Extensive support for rendering surfaces.
 Powerful 3D interactive molecule builder.
 Build & optimize structures for the entire periodic table.
 Support for spectroscopy, geometry optimizations, energies &
properties.
Step: 1: Open the Argus lab, Import the protein molecule.
Figure: 38: open the homepage.
70
Figure: 39: Open the structure.
Step:2 :the tree view of 3EQM & open up the residues/misc folder to
show the 3EQM residue.
Figure: 40: Tree view of 3EQM
71
Figure:41: Hide unselected structure.
Figure: 42 : Center molecule in window.
72
Figure: 43 : View/center molecule in window.
Figure: 44: Add hydrogen
73
Figure: 45: ctrl+c & ctrl+v the selected residue.
Figure: 46: Right click on 603ASD & select “make a ligand group
from this residue” .now group have two ligands name “1 ASD” &
“2ASD”.
74
Figure: 47: Right click on “1 ASD” in the group folder & select
“modify group” option. In modify group dialog box, type in a new
name like ‘lig’.
& “2 ASD” type in name like ‘ligand’.
Figure: 48: Right click on the group named “lig” & select the “make a
binding site group for this group”.
75
Figure: 49: Selecting the Calculation/DOCK a Ligand…..
Figure: 50: Settings dialog box looks like: select the ligand to dock in
the “Ligand “drop-box. Click on the “Calculate size”. Make sure
“ARGUS DOCK”. Than ok.
76
Figure: 51: See Argus Lab first generate the scoring grids used
during the docking…
Figure: 52: Go to calculation in the calculations folder select the
Dock a Database…
77
Figure: 53: Click on Get Ligands & select the sdf compound.
Figure: 54: See the result best Ligand phase-energy……..
78
3.1.2.4 Hex 4.5
Figure: 55: Open the homepage & go to file
Figure: 56: Hex after loading the molecule
open
receptor...
79
Figure: 57: Open the homepage & go to file
Figure: 58: Go to controls----- docking----- enter.
open
Ligand...
80
59: Docking control---activates ----hex progress.
Figure: 60: See the Hex progress….
81
Figure: 61: See the E total & rms…
Figure: 62 Save the result...
82
Figure: 63: Save both---- enter the name ---ok
3.1.2.5ADME Tox
http://pharma-algorithms.com/
ADME is an acronym in pharmacokinetics and
pharmacology for absorption, distribution, metabolism, and excretion,
and describes the disposition of a pharmaceutical compound within an
organism. The four criteria all influence the drug levels and kinetics of
drug exposure to the tissues and hence influence the performance and
pharmacological activity of the compound as a drug.
Absorption/Administration:-For a compound to reach a tissue, it
usually must be taken into the bloodstream - often via mucous surfaces
like the digestive tract (intestinal absorption) - before being taken up by
the target cells. This can be a serious problem at some natural barriers
like the blood-brain barrier. Factors such as poor compound solubility,
gastric emptying time, intestinal transit time, chemical instability in the
83
stomach, and inability to permeate the intestinal wall can all reduce the
extent to which a drug is absorbed after oral administration. Absorption
critically determines the compound's bioavailability. Drugs that absorb
poorly when taken orally must be administered in some less desirable
way, like intravenously or by inhalation (e.g. zanamivir).
Distribution:-The compound needs to be carried to its effectors site,
most often via the bloodstream. From there, the compound may
distribute into tissues and organs, usually to differing extents.
Metabolism:-Compounds begin to break down as soon as they enter the
body. The majority of small-molecule drug metabolism is carried out in
the liver by redox enzymes, termed cytochrome P450 enzymes. As
metabolism occurs, the initial (parent) compound is converted to new
compounds called metabolites. When metabolites are pharmacologically
inert, metabolism deactivates the administered dose of parent drug and
this usually reduces the effects on the body. Metabolites may also be
pharmacologically active, sometimes more so than the parent drug.
Excretion/Elimination:-Compounds and their metabolites need to be
removed from the body via excretion, usually through the kidneys
(urine) or in the feces. Unless excretion is complete, accumulation of
foreign substances can adversely affect normal metabolism.
Excretion of drugs by the kidney involves 3 main mechanisms:


Glomerular filtration of unbound drug.
Active secretion of (free & protein-bound) drug by transporters
e.g. anions such as urate, penicillin, glucuronide, sulfate
conjugates) or cations such as choline, histamine.
 Filtrate 100-fold concentrated in tubules for a favorable
concentration gradient so that it may be reabsorbed by passive
diffusion and passed out through the urine.
84
Toxicity:Sometimes, the potential or real toxicity of the compound is taken into
account (ADME-Tox or ADMET). When the Liberation of the substance
(from protective coating, or other excipients) is considered, we speak of
LADME.
Computational chemists try to predict the ADME-Tox qualities of
compounds through methods like QSPR or QSAR.The route of
administration critically influences ADME.
Find the ADME & TOX properties for the best molecules.
ADME Tox Box was opened using the URL:
http://pharma-algorithms.com/webboxes/
 Click on access free Demo ADME/Tox Web.
 Click on continue.
 Paste the canonical smile of the molecule or upload the files which
are saved in .mol extension & click on calculate.
 ADME/Tox Properties were displayed.
 Each of the properties were noted & studied.
Figure: 64: ADME Box
85
Figure: 65: TOX Box
Figure: 66: Docking score graph
86
4. Result
4. Result
4.1Table no.: 1. List of Aromatase inhibitor with Sources.
Sr.no. Compound name
1 Ascorbic Acid
Structure
Source
broccoligreen
red peppers,
collard greens
cauliflower
lemon
cabbage
87
2 Chrysin
Passiflora coerulea
passion flower
3 Phytoestrogens
Citrus
Tea
Wine
Dark chocolate
4 .Nicotine
Citrus
Tea
Wine
Dark chocolate
5 Quercetin
Citrus
Tea
Wine
Dark chocolate
Neem
6 Naringenin
Citrus
Tea
Wine
Dark chocolate
88
7 Resveratrol
Citrus
Tea
Wine
Dark chocolate
8 Apigenin
Citrus
Tea
Wine
Dark chocolate
Chamomile
9 Genistein
Citrus
Tea
Wine
Dark chocolate
10 Oleuropein
Citrus
Tea
Wine
Dark chocolate
Red clover
89
11 Astragalus
Astragalus
12
Aloe Vera
Aloin
13 Emodin
Aloe Vera
14 Campesterol
soybean oil
90
15 Lupeol
Olive
Mango
16 Sitosterol
Peanuts
17 Quinic acid
Green tea
18 Rutin
Broccoli
Broccoli
91
19 Sec-butyl isothiocyanate
Broccoli
20 Zeaxanthin
Tufuling
21 Poricoic acid
Tufuling
92
22
Zoledronic acid
SAN ANTONIO
23 Beta Carotene
Curcuma domestica
24 cyanocobalamin
Curcuma domestica
25 Ar-turmerone
Curcuma domestica
93
26 Zingiberene
Curcuma domestica
27 Cineol
Curcuma domestica
28 Borneol
Curcuma domestica
29 α-Phellandrene
Curcuma domestica
94
30 Sabinene
Curcuma domestica
31 ar-curcumene
Curcuma domestica
32 curcumol
Curcuma domestica
33 Vitamin D3
Salmon, Mackerel
95
34 Epigallocatechin gallate
Citrus
Tea
Wine
Dark chocolate
35 Vitamin E
Vegetable oils, nuts
36 Myristic acid
Fish oil
Agaricus bisporus
37 Palmitic acid
Fish oil
Agaricus bisporus
96
38 stearic acid
Fish oil
Agaricus bisporus
39 lauric acid
Fish oil
Agaricus bisporus
40 pentadecanoic acid
Fish oil
Agaricus bisporus
41 Allicin
Garlic
97
42
Ajoene
Garlic
43 Allylpropl
Garlic
44 Diallyl
Garlic
45 S-Allyl cysteine
Garlic
46 Ellagic Acid
strawberries
98
47 Folic Acid
Leafy vegetables
48 Catechin
Green tea
49 Theogallin
Green tea
50 Caffeine
Green tea
99
51 Theobromine
Green tea
52 Theophylline
Green tea
53 Thiamine
Green tea
54 Riboflavin
Green tea
100
55 Pyridoxine
Green tea
56 Gallic acid
Green tea
57 Melatonin
walnuts
58 Indole-3-carbinol
BROCCOLI
ORANGES
OATS
TOMATOES
BEANS
101
59 linoleic
corn
soybeans
sunflower seeds
canola oil
olive oil.
White button
mushrooms
60 α-Linolenic acid
corn
soybeans
sunflower seeds
canola oil
olive oil.
White button
mushrooms
61 oleic acids
Moringa oleifera
62 Linamarin
Cassava Latex
102
63 Azadirachtin
Neem
64 Nimbin
Neem
65 Gedunin
Neem
66 Salannin
Neem
103
67 Ryanodine
Noni
68 Daidzein
Red clover
69 Pratensein
Red clover
70 Methyl salicylate
Red clover
104
71 Eugenol
Red clover
72 Androsta-1,4,6-triene-3
,17-dione
73 Raffinose
Soya bean
74 stachyose
Soya bean
105
75 Stevioside
Stevia
76 steviol
Stevia
77 Amygdalin
Apples
Cherries
Peaches
Plums
Almonds
Papaya
Nectarines
78 Vitamin K
Grains and Nuts
Broccoli
106
79 Arachidonic acid
peanut oil
80 Pteropodine
Uncaria tomentosa
81 Rhynchophylline
Uncaria tomentosa
82 Hirsutine
Uncaria tomentosa
107
83 Quinovic acid glycosides
Uncaria tomentosa
84 Biotin
liver,
egg yolk,
cereals,
legumes and nuts
85 cinnamic acid
Bee propolis
Tobacco Leaves
86 Cinnamyl alcohol
Bee propolis
108
87 Vanillin
Bee propolis
88 Caffeic acid
Bee propolis
89 Isalpinin
Bee propolis
90 Pinocembrin
Bee propolis
109
91 Galangin
Bee propolis
92
Bee propolis
ferulic acid.
93 Galactose
dairy products
sugar beets
other gum
94 Rhamnose
dairy products
sugar beets
chlorell
110
95 lapachol
Pau d'arco
96 xyloidone
Pau d'arco
97 Sesquiterpene lactone
Sweet annie
98 Benzaldehyde
Gauva
111
99 Alpha-copaene
Gauva
100 Myrcene
Gauva
101 Hexyl acetate
Gauva
102 ethyl decanoate
Gauva
112
103 Alpha-humulene
Gauva
104 Ergosterol
Beetroot
105 Octacosanol
Beewax
106 Bisabolene
Ginger
113
107 Phellandrene
Ginger
108 Citral
Ginger
109 Citronellol
Ginger
110 linalool
Ginger
114
111 Limonene.
Ginger
112
Ginger
Zingiberol
113 Camphene
Ginger
114 Gingerol
Ginger
115
115 Shogaol
Ginger
116 Astaxanthin
Pacific krill
Antarctic krill
117 Phytol
Scutellaria Barbatae
118 wogonin
Scutellaria Barbatae
116
119 luteolin
Scutellaria Barbatae
120 hispidulin
Scutellaria Barbatae
121 Annotemoyin-2
Annona muricata
122 squamocin-I
Annona muricata
117
123 Squamocin O2
Annona muricata
124 Motrilin
Annona muricata
125 Squamostatin B
Annona muricata
126 cherimolin-1
Annona muricata
118
127 Solanine
Vernonia amygdalina
128 coumarine
Vernonia amygdalina
129 Vernodalin
Annona muricata
130 Vernolide
Annona muricata
119
131 Hydroxyvernolide
Annona muricata
132 Irofulven
Mushrooms
133 Kolavenol
Entada abyssinica
134 Alpha Pinene
Bergamot
120
135 Alpha Bergaptnen
Bergamot
136 Alpha Terpineol
Bergamot
137 Linalyl Acetate
Bergamot
138 Nerol
Bergamot
121
139 Neryl Acetate
Bergamot
140 Beta Bisabolene
Bergamot
141 Betulinic Acid
Betula alba
Eurya Japonica
142 Oenothein B
Epilobium
parviflorum
122
143 Cyclosporin A
Orange juice,Grape
juice
144 Midazolam
Orange juice,Grape
juice
145 Triazolam
Orange juice,Grape
juice
146 Terfenadine
Orange juice,Grape
juice
123
147 Saquinavir
Orange juice,Grape
juice
148 Ethynylestradiol
Orange juice,Grape
juice
149 Pseudolaric Acid B
Pseudolarix kaempferi
150 Indirubin
Chinese antileukaemia
124
151 Withaferin A
Withania somnifera
152 Morphine
Jasmin
153 Codeine
Jasmin
154 Colchine
Jasmin
125
155 Emetine
Jasmin
156 Rescinnamine
Jasmin
157 Ajmaline
Jasmin
158 Vinblastine
Jasmin
126
159 Vincristine
Jasmin
160 Eucalyptol
Ocimum basilicum
161 Camphor
Ocimum basilicum
162 Cis-ocimene
Ocimum basilicum
127
163 Estragole
Ocimum basilicum
164 Anethole
Fennel
165 farnesol
Lemon grass
166 Isovalerylaldehyde
Lemon grass
128
167 Decanaldehyde
Lemon grass
168 Pelletierine
punica granatum L
169 Rotenone
Jicama vine plant.
170 Arbutin
Damiana leaf
171 Hexacosanol-1
Damiana leaf
129
4.2Table no.: 2. List of Aromatase inhibitor with compound
properties:
Sr.no.
1
Mol.weight
176.12412 [g/mol]
2
254.2375 [g/mol]
3
273.255385 [g/mol]
4
Mol.formula
C6H8O6
XlogP
1.8
H.B.Donor H.b.Acceptor
4
6
1
2
4
C15H10O4
C15H10O5
2.7
3
5
162.23156 [g/mol]
C10H14N2
1.2
0
2
5
302.2357 [g/mol]
C15H10O7
1.5
5
7
6
272.25278 [g/mol]
C15H12O5
2.4
3
5
7
228.24328 [g/mol]
C14H12O3
3.1
3
3
8
270.2369 [g/mol]
C15H10O5
1.7
3
5
9
270.2369 [g/mol]
C15H10O5
2.7
3
10
540.51378 [g/mol]
C25H32O13
0.4
6
13
11
321.21052 [g/mol]
C19H13Br
6.2
0
0
12
418.39398 [g/mol]
C21H22O9
2.2
7
9
13
270.2369 [g/mol]
C15H10O5
2.7
3
5
14
400.68012 [g/mol]
C28H48O
8.8
1
1
15
426.7174 [g/mol]
C30H50O
9.9
1
1
16
414.7067 [g/mol]
C29H50O
9.3
1
1
17
91.15864 [g/mol]
C7H11O6
-1.7
4
6
18
610.5175 [g/mol]
C27H30O16
-1.3
10
16
19
115.19666 [g/mol]
C5H9NS
2.8
0
1
130
20
568.87144 [g/mol]
C40H56O2
10.9
2
2
21
498.69394 [g/mol]
C31H46O5
6.4
3
5
22
272.089622[g/mol]
C5H10N2O7P2
-4.3
5
8
23
536.87264 [g/mol]
C40H56
13.5
0
0
24
1355.365181 [g/mol]
C63H88CoN14O14P
9
21
25
216.3187 [g/mol]
C15H20O
4
0
1
26
204.35106 [g/mol]
C15H24
5.2
0
0
27
154.24932 [g/mol]
C10H18O
2.5
0
1
28
154.24932 [g/mol]
C10H18O
2.7
1
1
29
136.23404 [g/mol]
C10H16
3.2
0
0
30
136.23404 [g/mol]
C10H16
3.1
0
0
31
202.33518 [g/mol]
C15H22
5.4
0
0
32
236.34986 [g/mol]
C15H24O2
2.8
1
2
33
384.63766 [g/mol]
C27H44O
7.9
1
1
34
458.37172 [g/mol]
C22H18O11
1.2
8
11
35
430.7061 [g/mol]
C29H50O2
10.7
1
2
36
228.37092 [g/mol]
C14H28O2
5.3
1
2
37
256.42408 [g/mol]
C16H32O2
6.4
1
2
38
284.47724 [g/mol]
C18H36O2
7.4
1
2
39
200.31776 [g/mol]
C12H24O2
4.2
1
2
40
242.3975 [g/mol]
C15H30O2
5.8
1
2
41
162.273 [g/mol]
C6H10OS2
1.3
0
1
42
234.40186 [g/mol]
C9H14OS3
1.7
0
1
131
43
148.28948 [g/mol]
C6H12S2
2.4
0
0
44
146.2736 [g/mol]
C6H10S2
2.2
0
0
45
161.22204 [g/mol]
C6H11NO2S
2.1
2
3
46
302.19264 [g/mol]
C14H6O8
1.1
4
8
47
441.39746 [g/mol]
C19H19N7O6
1.1
6
11
48
290.26806 [g/mol]
C15H14O6
0.4
5
6
49
344.27084 [g/mol]
C14H16O10
1.2
7
10
50
194.1906 [g/mol]
C8H10N4O2
0.1
0
3
51
180.16402 [g/mol]
C7H8N4O2
0.8
1
3
52
180.16402 [g/mol]
C7H8N4O2
0
1
3
53
265.35458 [g/mol]
C12H17N4OS
1
2
4
54
376.3639 [g/mol]
C17H20N4O6
1.5
5
8
55
169.17784 [g/mol]
C8H11NO3
0.8
3
4
56
170.11954 [g/mol]
C7H6O5
0.7
4
5
57
232.27834 [g/mol]
C13H16N2O2
0.8
2
2
58
147.17386 [g/mol]
C9H9NO
1.1
2
1
59
280.44548 [g/mol]
C18H32O2
6.8
1
2
60
278.4296 [g/mol]
C18H30O2
5.9
1
2
61
282.46136 [g/mol]
C18H34O2
6.5
1
2
62
247.24508 [g/mol]
C10H17NO6
1.8
4
7
63
720.71426 [g/mol]
C35H44O16
-0.6
3
16
64
540.60144 [g/mol]
C30H36O9
2.3
0
9
132
65
484.58124 [g/mol]
C28H36O7
4
0
7
66
596.70776 [g/mol]
C34H44O9
3.9
0
9
67
493.5467 [g/mol]
C25H35NO9
-0.5
7
9
68
254.2375 [g/mol]
C15H10O4
2.5
2
4
69
300.26288 [g/mol]
C16H12O6
2.6
3
6
70
152.14732 [g/mol]
C8H8O3
2.3
1
3
71
164.20108 [g/mol]
C10H12O2
2
1
2
72
282.37678 [g/mol]
C19H22O2
3
0
2
73
504.43708 [g/mol]
C18H32O16
5.8
11
16
74
666.57768 [g/mol]
C24H42O21
8
14
21
75
804.8722 [g/mol]
C38H60O18
-1.2
11
18
76
318.4504 [g/mol]
C20H30O3
3.8
2
3
77
457.42848 [g/mol]
C20H27NO11
-2.7
7
12
78
450.69574 [g/mol]
C31H46O2
10.9
0
2
79
304.46688 [g/mol]
C20H32O2
6.3
1
2
80
368.42626 [g/mol]
C21H24N2O4
1.6
1
5
81
384.46872 [g/mol]
C22H28N2O4
2.3
1
5
133
82
83
368.46932 [g/mol]
486.68324 [g/mol]
C22H28N2O3
C30H46O5
3.4
5.9
1
3
4
5
84
244.31064 [g/mol]
C10H16N2O3S
0.3
3
3
85
148.15862 [g/mol]
C9H8O2
2.1
1
2
86
134.1751 [g/mol]
C9H10O
1.9
1
1
87
152.14732 [g/mol]
C8H8O3
1.2
1
3
88
89
180.15742 [g/mol]
284.26348 [g/mol]
C9H8O4
C16H12O5
1.2
2.6
3
2
4
5
90
256.25338 [g/mol]
C15H12O4
2.7
2
4
91
270.2369 [g/mol]
C15H10O5
2.3
3
5
92
194.184 [g/mol]
C10H10O4
1.5
2
4
93
180.15588 [g/mol]
C6H12O6
-2.6
5
6
94
164.15648 [g/mol]
C6H12O5
-2.1
4
5
95
242.26986 [g/mol]
C15H14O3
2.8
1
3
96
240.25398 [g/mol]
C15H12O3
2.6
0
3
97
264.3169 [g/mol]
C15H20O4
1.3
1
4
98
106.12194 [g/mol]
C7H6O
1.5
0
1
99
204.35106 [g/mol]
C15H24
4.5
0
0
100
136.23404 [g/mol]
C10H16
4.3
0
0
101
144.21144 [g/mol]
C8H16O2
2.4
0
2
102
200.31776 [g/mol]
C12H24O2
4.6
0
2
103
204.35106 [g/mol]
C15H24
4.5
0
0
104
517.55478 [g/mol]
C23H27N5O7S
30.5
5
7
105
410.75952 [g/mol]
C28H58O
3.8
1
1
106
204.35106 [g/mol]
C15H24
4.7
0
0
107
136.23404 [g/mol]
C10H16
3.2
0
0
134
108
152.23344 [g/mol]
C10H16O
3
0
1
109
156.2652 [g/mol]
C10H20O
3.2
1
1
110
154.24932 [g/mol]
C10H18O
2.7
1
1
111
136.23404 [g/mol]
C10H16
3.4
0
0
112
236.39292 [g/mol]
C16H28O
4.5
1
1
113
136.23404 [g/mol]
C10H16
3.3
0
0
114
294.38594 [g/mol]
C17H26O4
3.1
2
4
115
276.37066 [g/mol]
C17H24O3
3.7
1
3
116
596.83848 [g/mol]
C40H52O4
10.3
2
4
117
296.531 [g/mol]
C20H40O
8.2
1
1
118
284.26348 [g/mol]
C16H12O5
3
2
5
119
286.2363 [g/mol]
C15H10O6
1.4
4
6
120
300.26288 [g/mol]
C16H12O6
1.7
3
6
121
312.4517 [g/mol]
C14H15KN3OS
0
2
2
122
638.91514 [g/mol]
C37H66O8
8
4
8
123
638.91514 [g/mol]
C37H66O8
8
4
8
124
622.91574 [g/mol]
C37H66O7
9.3
3
7
125
638.91514 [g/mol]
C37H66O8
8
4
8
126
258.31386 [g/mol]
C19H14O
4
1
1
127
868.05882 [g/mol]
C45H73NO15
1.8
9
16
128
146.14274 [g/mol]
C9H6O2
1.4
0
2
129
360.3579 [g/mol]
C19H20O7
1.6
1
7
135
130
362.37378 [g/mol]
C19H22O7
0.9
1
7
131
378.37318 [g/mol]
C19H22O8
-0.3
2
8
132
246.30162 [g/mol]
C15H18O3
-0.2
2
3
133
290.48336 [g/mol]
C20H34O
6
1
1
134
135
136.23404 [g/mol]
216.1895 [g/mol]
C10H16
C12H8O4
2.8
0
0
136
154.24932 [g/mol]
C10H18O
1.8
1
1
137
196.286 [g/mol]
C12H20O2
3.3
0
2
138
154.24932 [g/mol]
C10H18O
2.9
1
1
139
196.286 [g/mol]
C12H20O2
3.5
0
2
140
204.35106 [g/mol]
C15H24
5.2
0
0
141
456.70032 [g/mol]
C30H48O3
8.2
2
3
142
1571.0982 [g/mol]
C68H50O44
1.3
25
44
143
1202.61124 [g/mol]
C62H111N11O12
7.5
5
12
144
325.767323 [g/mol]
C18H13ClFN3
2.5
0
3
145
343.20998 [g/mol]
C17H12Cl2N4
2.4
0
3
146
471.67344 [g/mol]
C32H41NO2
6.6
2
3
147
670.8408 [g/mol]
C38H50N6O5
4.2
5
7
148
296.40336 [g/mol]
C20H24O2
3.7
2
2
136
149
432.46362 [g/mol]
C23H28O8
2.3
1
8
150
262.2628 [g/mol]
C16H10N2O2
2.3
2
3
151
470.59772 [g/mol]
C28H38O6
3.8
2
6
152
285.33766 [g/mol]
C17H19NO3
0.8
2
4
153
299.36424 [g/mol]
C18H21NO3
1.1
1
4
154
399.437 [g/mol]
C22H25NO6
1
1
6
155
480.6389 [g/mol]
C29H40N2O4
4.7
1
6
156
634.71598 [g/mol]
C35H42N2O9
4.5
1
10
157
326.43264 [g/mol]
C20H26N2O2
1.8
2
4
158
909.0526 [g/mol]
C46H60N4O13S
4.711
5
16
159
824.95764 [g/mol]
C46H56N4O10
2.8
3
12
160
154.24932 [g/mol]
C10H18O
2.5
0
1
161
152.23344 [g/mol]
C10H16O
2.2
0
1
162
136.23404 [g/mol]
C10H16
4.3
0
0
163
148.20168 [g/mol]
C10H12O
3.4
0
1
164
148.20168 [g/mol]
C10H12O
3.3
0
1
165
222.36634 [g/mol]
C15H26O
4.8
1
1
166
86.1323 [g/mol]
C5H10O
1
0
1
167
168
156.2652 [g/mol]
141.2108 [g/mol]
C10H20O
C8H15NO
3.8
0.4
0
1
1
2
169
394.41718 [g/mol]
C23H22O6
4.1
0
6
170
272.25124 [g/mol]
C12H16O7
0.7
5
7
171
382.70636 [g/mol]
C26H54O
12.7
1
1
137
4.3Table no.:3. Quantum, Argus Lab & Hex Docking score:
No.
Name
1 Aminoglutethimide
2 Testolactone (Teslac)
Anastrozole
3 (Arimidex)
4 Letrozole (Femara)
Exemestane
5 (Aromasin)
6 Vorozole (Rivizor)
Formestane
7 (Lentaron)
8 Fadrozole (Afema)
9
3EQM
1
2
3
4
5
6
7
8
9
10
3EQM
11
12
13
14
15
16
17
18
19
1,4,6-androstatrien-3,
17-dione (ATD)
Ascorbic Acid
Chrysin
Phytoestrogens
.Nicotine
Quercetin
Naringenin
Resveratrol
Apigenin
Genistein
Oleuropein
Astragalus
Aloin
Emodin
Campesterol
Lupeol
Sitosterol
Quinic acid
Rutin
Sec-butyl
Quantum
HEX
G value
RMS
E value RMS
-17.57
106.24
-19.95
-1
Argus
lab
-19.6
94.67
-9.87
-18.61
-24.3
96.73
108.85
-23.84
-11.46
-9.2672
-1 10.6252
-1 12.6331
-1 -13.653
-16.32
-20.47
94.67
96.75
-22.24
-17.07
-1
-1
-9.3331
-10.88
-10.37
-23.59
95.66
111.02
-15.75
-11.24
-1
-1
-10.38
-12.95
-26.12
-21.56
-18.7
-21.6
-20.16
-15.73
-23.37
-21.22
-25.08
-22.82
-20.42
-18.2
-22.17
-29.5
-26.01
-25.47
-39.4
-24.5
-21.75
-18.41
-23.69
-13.13
105.86
19.97
110.05
104.61
97.09
93.55
109.47
110.97
105.83
105.37
108.21
102.4
6.37
107.07
111.15
112
112.58
97.55
99.18
107.15
100.41
99.08
-14.6
-1
-14.83
-15.08
-15.27
-15.27
-8.3
-10.65
-9.62
-15.21
-14.93
-22.61
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-10.98
-12.718
-6.59
-10.278
-12.422
-7.34
-9.55
-9.71
-11.46
-10.35
-10.02
0
-27.66
-30.52
-18.43
-22.03
-18.72
-20.99
0
-23.76
-11.69
-1
-1
-1
-1
-1
-1
-1
-1
-1
-14.72
0
-10.85
0
0
0
-7.97
0
-6.56
Lipinsky
rule
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Yes
No
No
No
Yes
No
Yes
138
isothiocyanate
20
3EQM
21
22
23
24
25
26
27
28
29
30
3EQM
31
32
33
34
35
36
37
38
39
40
3EQM
41
42
43
44
45
46
47
48
49
50
3EQM
51
52
Zeaxanthin
Poricoic acid
Zoledronic acid
Beta Carotene
cyanocobalamin
Ar-turmerone
Zingiberene
Cineol
Borneol
α-Phellandrene
Sabinene
ar-curcumene
curcumol
Vitamin D3
Epigallocatechin
gallate
Vitamin E
Myristic acid
Palmitic acid
stearic acid
lauric acid
pentadecanoic acid
Allicin
Ajoene
Allylpropl
Diallyl
S-Allyl cysteine
Ellagic Acid
Folic Acid
Catechin
Theogallin
Caffeine
Theobromine
Theophylline
Fail
-21.2
Fail
-30.24
Fail
Fail
-19.67
-19.86
-15.95
-13.95
Fail
-11.87
-13.29
-24.34
-21.18
-0.91
Fail
21.87
Fail
93.97
Fail
Fail
95.59
98.41
97.32
95.94
Fail
93.01
8.63
112.74
107.98
106.9
-15.75
-1
0
No
-24.7
-16.26
-16.26
-34.96
0
-19.41
0
-19.29
Fail
0
-1
-1
-1
-1
-1
-1
-1
-1
Fail
-1
0
0
0
0
-11.878
-12.27
-8.79
-9.25
0
-9.8
Yes
No
No
No
Yes
Yes
Yes
Yes
Yes
Yes
-11.36
-14.27
-18.09
-1
-1
-1
-12.06
-11.2
0
Yes
Yes
No
-34.27
-15.27
-29.29
-30.78
-26.55
-28.93
112.85
107.04
107.17
115.89
103.72
104.59
-20.48
-11.83
-8.56
-7.98
-6.91
-10.29
-1
-1
-1
-1
-1
-1
No
No
Yes
No
No
Yes
-29.92
-19.86
-14.03
-17.33
-13.2
-12.32
-15.33
-20.41
-26.78
-16.58
-21.75
-11.04
-19.28
-16.32
-17.24
109.48
20.55
92.16
101.13
97.22
99.13
97.58
99.57
101.68
99.67
97.55
94.75
20.17
107.74
108.28
-5.94
-1
0
0
-11.224
-11.93
-12.674
-10.342
116276
-8.64
-10.83
-15.03
-14.13
-7.73
-14.5
-21.42
-10.82
-25.67
-12.95
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-7.7659
-10.599
-8.3388
-9.0611
-7.8112
-8.7216
-9.8648
-9.9197
-9.4515
-4.8859
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Yes
-9.47
-10.55
-1
-1
-4.9326
-4.9077
Yes
Yes
No
139
53
54
55
56
57
58
59
60
3EQM
61
62
63
64
65
66
67
68
69
70
3EQM
71
Thiamine
Riboflavin
Pyridoxine
Gallic acid
Melatonin
Indole-3-carbinol
linoleic
α-Linolenic acid
oleic acids
Linamarin
Azadirachtin
Nimbin
Gedunin
Salannin
Ryanodine
Daidzein
Pratensein
Methyl salicylate
Eugenol
Androsta-1,4,6-triene3
,17-dione
Raffinose
stachyose
Stevioside
steviol
Amygdalin
Vitamin K
Arachidonic acid
Pteropodine
72
73
74
75
76
77
78
79
80
3EQM
81 Rhynchophylline
82 Hirsutine
Quinovic acid
83 glycosides
84 Biotin
85 cinnamic acid
-24.94
104.65
-5.14
-1
-32.14
110.37
-16.96
-1
-18.72
105.68
-15.19
-1
-20.78
107.19
-15.04
-1
-18.97
110.83
-17.48
-1
-16.42
107.68
-13.71
-1
-31.08
108.22
-18.67
-1
Stopped Stopped stopped Stopped
8.46
7.36
-29.86
116.52
-9.01
-1
-13.41
105.28
-10.39
-1
Fail
Fail
-19.73
-1
-8.8
105.5
-32.33
-1
-18.52
85.75
-18.34
-1
-19.14
102.26
-15.34
-1
-16.78
98.79
-31.17
-1
-27.42
105.96
-16.03
-1
-27.29
112.45
-13.16
-1
-18
108.85
-7.48
-1
-18.26
5.67
-17.61
106.05
-16.2
-1
-8.6123
-7.1596
-6.9589
-8.2819
-8.5331
-8.1227
-14.727
0
Yes
Yes
Yes
Yes
Yes
Yes
No
No
0
-7.7826
0
0
0
0
0
-9.7565
-9.5339
-8.1565
Yes
Yes
No
No
Yes
No
Yes
Yes
Yes
No
-9.4711
Yes
-13.56
-5.2
14.54
676.96
13.68
-8.64
-26.19
-24.64
-27.13
-25.72
-25.35
-21.86
96.08
113.64
110.35
106.43
115.02
110.04
111.69
115.42
110.87
5.15
109.5
95.14
-13.98
-26.76
-22.94
-34.05
-19.42
-21.47
-12.42
-14.42
-14.41
-1
-1
-1
-1
-1
-1
-1
-1
-1
-12.633
0
0
0
-13.525
0
0
-14.944
0
Yes
No
No
No
Yes
No
Yes
Yes
Yes
-8.5
0
-1
-1
-8.9502
-10.133
Yes
Yes
-21.54
-30.56
-22.39
94.35
111.71
102.59
-20.33
-15.75
-17.9
-1
-1
-1
0
-9.0323
-9.6275
No
Yes
Yes
140
86
87
88
89
90
3EQM
91
92
93
94
95
96
97
98
Cinnamyl alcohol
Vanillin
Caffeic acid
Isalpinin
Pinocembrin
-15.81
-16.78
-24.9
-20.98
-19.23
-17.03
-21.57
-24.07
-15.09
-16.23
-22.55
-19.22
-20.35
-13
102.72
103.27
112.03
102.11
105.93
19.05
108.3
109.71
108.35
111.31
114.29
111.92
96.81
111.72
-18.97
0
-17.73
-9.43
-13.11
-1
-1
-1
-1
-1
-9.1332
-7.6467
-9.3408
-9.7448
-10.058
Yes
Yes
Yes
Yes
Yes
0
-8.46
-9.57
0
-14.98
-9.58
-31.43
-11.13
-1
-1
-1
-1
-1
-1
-1
-1
-10.005
-8.9502
-6.1938
-6.7902
-11.544
-10.43
-10.272
-8.3353
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
-17.43
-15.51
-25.57
108.64
113.82
5.74
-9.41
-7.37
-1
-1
-8.3353
-12.14
Yes
Yes
-15.66
113.97
-24.07
-1
-9.9604
Yes
109.49
106.31
110.14
Fail
109.5
112.43
111.7
109.54
109.14
15.06
115.65
-5.81
-15.14
-14.84
-11.24
-14.27
-21.56
-10.74
0
0
-1
-1
-1
-1
-1
-1
-1
-1
-1
######
-12.105
0
-12.729
-12.133
-9.575
-9.5945
-9.2665
9.475
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
Limonene.
-21.39
-16.2
-27.28
Fail
-20.58
-15.62
-18.5
-18.53
-11.58
-19.16
-12.72
-24.07
-1
-9.4172
Yes
Zingiberol
Camphene
Gingerol
Shogaol
Astaxanthin
-16.92
-12.28
-20.46
-22.34
Fail
112.56
122.31
111.92
123.8
Fail
-16.41
-19.97
-18.32
-14.41
-15.57
-1
-1
-1
-1
-1
-11.34
-9.5637
-10.843
-11.829
0
Yes
Yes
Yes
Yes
No
Galangin
ferulic acid.
Galactose
Rhamnose
lapachol
xyloidone
Sesquiterpene lactone
Benzaldehyde
99 Alpha-copaene
100 Myrcene
3EQM
Hexyl acetate
101
102
103
104
105
106
107
108
109
110
3EQM
111
112
113
114
115
116
ethyl decanoate
Alpha-humulene
Ergosterol
Octacosanol
Bisabolene
Phellandrene
Citral
Citronellol
Citral
141
117
118
119
120
3EQM
121
122
123
124
125
126
127
128
129
130
3EQM
131
132
133
134
135
136
137
138
139
140
3EQM
141
142
143
144
145
146
147
148
149
150
3EQM
151
Phytol
wogonin
luteolin
hispidulin
Annotemoyin-2
squamocin-I
Squamocin O2
Motrilin
Squamostatin B
cherimolin-1
Solanine
coumarine
Vernodalin
Vernolide
Hydroxyvernolide
Irofulven
Kolavenol
Alpha Pinene
Alpha Bergaptnen
Alpha Terpineol
Linalyl Acetate
Nerol
Neryl Acetate
Beta Bisabolene
Betulinic Acid
Oenothein B
Cyclosporin A
Midazolam
Triazolam
Terfenadine
Saquinavir
Ethynylestradiol
Pseudolaric Acid B
Indirubin
Withaferin A
-25.42
111.91
-20.5
112.9
-23.3
110.9
-16.7
113.69
-17.62
18.44
-22.82
96.36
Fail
Fail
Fail
Fail
Fail
Fail
Fail
Fail
Fail
Fail
-21.89
108.69
Fail
Fail
Stopped Stopped
Fail
Fail
-17.75
20.12
-27.59
108.6
-13.78
94.69
-22.92
111.57
-12.7
95.53
-15.64
98.58
-15.39
109.3
-17.84
96.68
-20.73
105.72
-32.67
102.76
-24.84
103.96
-12.54
3.33
Fail
Fail
Fail
Fail
Fail
Fail
-20.97
109.69
-22.4
108.29
-16.1
113.27
60.42
106.22
-23.17
113.63
-21.52
109.37
-23.21
102.91
-22.61
6.05
41.93
106.19
-12.12
-16.32
-13.93
-12.61
-1
-1
-1
-1
-12.187
-9.7062
-9.8326
-6.604
No
Yes
Yes
No
-17.71
-23.1
-23.1
-16.79
-30.49
0
-22.82
-12.14
-14.6
-17.25
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-10.094
0
0
0
0
-14.406
0
-8.7796
0
10.895
Yes
No
No
No
No
Yes
No
Yes
Yes
Yes
-19.39
-12.14
-10.57
-9.63
-20.48
-20.48
-12.54
-8.59
-10.27
-9.76
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-9.5551
-11.093
-13.951
-9.1635
-9.6195
-9.6195
9.7154
-9.5316
-10.342
-12.418
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
Yes
No
-22.03
-28.72
-25.17
-18.9
-16.06
-11.44
-31.81
-10.82
-25.67
-14.32
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
0
0
0
-11.158
-12.5
0
0
-9.9196
-9.4515
-10.25
Yes
No
No
Yes
Yes
No
No
Yes
Yes
Yes
-23.21
-1
0
Yes
142
152
153
154
155
156
157
158
159
160
3EQM
161
162
163
164
165
166
167
168
169
170
171
Morphine
Codeine
Colchine
Emetine
Rescinnamine
Ajmaline
Vinblastine
Vincristine
Eucalyptol
Camphor
Cis-ocimene
Estragole
Anethole
farnesol
Isovalerylaldehyde
Decanaldehyde
Pelletierine
Rotenone
Arbutin
Hexacosanol-1
-19.3
4.26
-19.08
19.87
54.32
Fail
Fail
-18.72
-15.45
-12.96
-12.75
-17.57
-17.19
-18.38
-18.87
-12.56
-17.12
-14.95
-22.41
-24.85
Fail
114.19
112.09
109.45
102.94
102.65
Fail
Fail
106.75
103.2
5.66
111
112.37
108.23
112.61
115.58
116.94
107.71
109.79
105.1
104.41
Fail
-18.36
-18.72
-25.01
-16.1
-30.7
-12.83
-27.32
-22.9
-9.2
-1
-1
-1
-1
-1
-1
-1
-1
-1
-10.649
0
-8.947
0
0
-11.446
0
-14.72
-8.7985
Yes
Yes
Yes
Yes
No
Yes
No
No
Yes
-16.77
-16.2
-21.27
-20.48
-15.32
-17.59
0
-28.75
-16.13
-16.5
-7.7
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-1
-9.5953
-9.8392
-9.364
-9.5443
-12.464
7
-9.0682
-7.403
0
-9.2137
-14.102
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
4.4Table no.:4 Docking score:
Sr.no
No.
1
2
3
4
5
6
7
8
Name
Aminoglutethimide
Testolactone (Teslac)
Anastrozole
(Arimidex)
Letrozole (Femara)
Exemestane
(Aromasin)
Vorozole (Rivizor)
Formestane
(Lentaron)
Fadrozole (Afema)
1,4,6-androstatrien-3,
9 17-dione (ATD)
Quantum
-17.57
-19.6
-18.61
-24.3
-16.32
-20.47
-10.37
-23.59
-26.12
143
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
1
2
3
5
6
7
8
9
11
13
25
36
39
46
53
54
56
61
68
69
81
82
84
85
91
92
97
115
139
150
169
170
Ascorbic Acid
Chrysin
Phytoestrogens
Quercetin
Naringenin
Resveratrol
Apigenin
Genistein
Astragalus
Emodin
Ar-turmerone
Myristic acid
lauric acid
Ellagic Acid
Thiamine
Riboflavin
Gallic acid
oleic acids
Daidzein
Pratensein
Rhynchophylline
Hirsutine
Biotin
cinnamic acid
Galangin
ferulic acid.
Sesquiterpene lactone
Shogaol
Neryl Acetate
Indirubin
Rotenone
Arbutin
-18.7
-21.6
-20.16
-23.37
-21.22
-25.08
-22.82
-20.42
-29.5
-25.47
-19.67
-29.29
-28.93
-20.41
-24.94
-32.14
-20.78
-29.86
-27.42
-27.29
-25.35
-21.86
-30.56
-22.39
-21.57
-24.07
-20.35
-22.34
-32.67
-23.21
-22.41
-24.85
144
4.5Table no.:5. IC50 Result:
Sr.no No.
1
1
2
2
3
3
4
5
5
6
6
7
7
8
8
9
9
11
10
13
11
25
12
36
13
39
14
46
15
53
16
54
17
56
18
61
19
68
20
69
21
81
22
82
23
84
24
85
25
91
26
92
27
28
29
30
31
32
97
115
139
150
169
170
Name
Ascorbic Acid
Chrysin
Phytoestrogens
Quercetin
Naringenin
Resveratrol
Apigenin
Genistein
Astragalus
Emodin
Ar-turmerone
Myristic acid
lauric acid
Ellagic Acid
Thiamine
Riboflavin
Gallic acid
oleic acids
Daidzein
Pratensein
Rhynchophylline
Hirsutine
Biotin
cinnamic acid
Galangin
ferulic acid.
Sesquiterpene
lactone
Shogaol
Neryl Acetate
Indirubin
Rotenone
Arbutin
Quantum
IC50
e value
-18.7
9.83
-1
-21.6
1.88
1
-20.16
6.45
2
-23.37
6.86
-1
-21.22
2.09
-1
-25.08
3.43
-1
-22.82
2.65
-1
-20.42
3.6
-1
-29.5
1.05
0
-25.47
1.78
0
-19.67
8.31
-1
-29.29
1.75
0
-28.93
2.37
0
-20.41
1.11
0
-24.94
2.56
-2
-32.14
2.97
-1
-20.78
9.89
-1
-29.86
3.45
0
-27.42
4.13
-1
-27.29
5.1
-6
-25.35
1.39
0
-21.86
9.08
-1
-30.56
2.74
-1
-22.39
1.17
0
-21.57
3.38
-1
-24.07
9.88
-1
-20.35
-22.34
-32.67
-23.21
-22.41
-24.85
5.25
2.32
1.9
1.53
6.25
2.29
0
0
0
0
-1
0
145
4.6Table no.:6 ADME-Tox properties:
S.
N
o
MOLECULE
ADME Properties
Oral
bioavailabi
lity
1
Ascorbic
Acid
2
Chrysin
3
Phytoestrog
ens
4
Quercetin
5
Naringenin
Probability
that
compound
has:
%F(Oral) >
30%: 0.033
%F(Oral) >
70%: 0.008
Probability
that
compound
has:
%F(Oral) >
30%: 0.811
%F(Oral) >
70%: 0.756
Probability
that
compound
has:
%F(Oral) >
30%: 0.849
%F(Oral) >
70%: 0.409
Probability
that
compound
has:
%F(Oral) >
30%: 0.033
%F(Oral) >
70%: 0.008
Probability
that
compound
has:
%F(Oral) >
30%: 0.350
%F(Oral) >
70%: 0.049
Solu
bility
-1.65
TOXICITY Properties
Drug
binding
to
plasma
protein
vd
(volum
e of
distrib
ution)
AME
S
%PPB:
98.22%
0.34
L/kg
Health effect
B*
C*
G*
K*
Li*
Lu*
0.960
0.78
0.36
0.61
0.07
0.12
0.91
0.65
L/kg
0.549
0.48
0.5
4
0.2
7
0.5
2
0.3
5
0.2
7
0.64
L/kg
0.194
0.46
0.61
0.46
0.59
0.51
0.25
0.60
L/kg
0.69
0.69
0.27
0.41
0.54
0.09
0.38
0.65
L/kg
0.072
0.34
0.66
0.48
0.81
0.32
0.27
LogKaHSA:
3.49
-3.37
%PPB:
87.10%
LogKaHSA
: 4.11
-3.38
%PPB:
87.87%
LogKaHSA
: 3.91
-3.88
%PPB:
93.68%
LogKaHSA
: 3.92
-3.10
%PPB:
86.32%
LogKaHSA
: 3.89
146
%PPB:
76.03%
6
-3.09
LogKaHSA:
3.76
%PPB:
76.03%
0.76
L/kg
0.124
0.56
0.50
0.36
0.13
0.53
0.13
0.64
L/kg
0.309
0.71
0.38
0.06
0.46
0.34
0.25
0.64
L/kg
0.194
0.46
0.61
0.46
0.59
0.51
0.25
%PPB:
99.86%
%PPB:
99.86%
1.000
0.94
0.82
0.98
0.63
0.90
0.98
LogKaHSA
: 6.05
LogKaHS
A
: 6.05
%PPB:
93.90%
0.92
L/kg
0.991
0.19
0.57
0.56
0.21
0.33
0.68
2.15
L/kg
0.299
0.21
0.33
0.08
0.17
0.15
0.20
0.40
L/kg
0.009
0.15
0.08
0.05
0.11
0.06
0.04
0.38
L/kg
0.015
0.14
0.19
0.06
0.04
0.05
0.05
LogKaHSA
: 3.76
Resveratrol
7
Apigenin
8
Genistein
9
Astragalus
10
Emodin
11
Arturmerone
12
Myristic
acid
13
lauric acid
Probability
that
compound
has:
%F(Oral) >
30%: 0.849
%F(Oral) >
70%: 0.409
Probability
that
compound
has:
%F(Oral) >
30%: 0.849
%F(Oral) >
70%: 0.409
Probability
that
compound
has:
%F(Oral) >
30%: 0.811
%F(Oral) >
70%: 0.205
Probability
that
compound
has:
%F(Oral) >
30%: 0.722
%F(Oral) >
70%: 0.209
Probability
that
compound
has:
%F(Oral) >
30%: 0.811
%F(Oral) >
70%: 0.302
Probability
that
compound
has:
%F(Oral) >
30%: 0.854
%F(Oral) >
70%: 0.450
Probability
that
compound
has:
%F(Oral) >
30%: 0.854
-3.35
%PPB:
87.87%
LogKaHSA
: 3.91
-3.38
%PPB:
87.87%
LogKaHSA
: 3.91
-7.31
-3.54
LogKaHSA
: 4.34
-3.73
%PPB:
92.71%
LogKaHSA
: 3.84
-5.68
%PPB:
99.02%
LogKaHSA
: 4.87
-4.38
%PPB:
96.77%
LogKaHSA
: 4.54
147
%F(Oral) >
70%: 0.450
14
Ellagic Acid
15
Thiamine
16
Riboflavin
17
Gallic acid
18
oleic acids
19
Daidzein
20
Pratensein
Probability
that
compound
has:
%F(Oral) >
30%: 0.033
%F(Oral) >
70%: 0.010
Probability
that
compound
has:
%F(Oral) >
30%: 0.363
%F(Oral) >
70%: 0.097
Probability
that
compound
has:
%F(Oral) >
30%: 0.479
%F(Oral) >
70%: 0.193
Probability
that
compound
has:
%F(Oral) >
30%: 0.033
%F(Oral) >
70%: 0.010
Probability
that
compound
has:
%F(Oral) >
30%: 0.854
%F(Oral) >
70%: 0.450
Probability
that
compound
has:
%F(Oral) >
30%: 0.811
%F(Oral) >
70%: 0.756
Probability
that
compound
has:
%F(Oral) >
30%: 0.722
%F(Oral) >
70%: 0.248
-3.63
%PPB:
88.99%
0.61
L/kg
0.391
0.90
0.27
0.95
0.95
0.83
0.73
0.96
L/kg
0.384
0.47
0.47
0.87
0.78
0.19
0.66
0.76
L/kg
0.182
0.88
0.90
0.91
0.53
0.91
0.66
0.29
L/kg
0.157
0.25
0.06
0.21
0.09
0.08
0.09
0.49
L/kg
0.011
0.19
0.17
0.06
0.21
0.10
0.49
1.03
L/kg
0.057
0.48
0.54
0.45
0.58
0.35
0.29
0.63
L/kg
0.448
0.40
0.86
0.77
0.80
0.41
0.20
LogKaHSA
: 3.61
-3.76
%PPB:
9.97%
LogKaHSA
: 1.46
-3.00
%PPB:
56.33%
LogKaHSA
: 2.44
-1.02
%PPB:
15.40%
LogKaHSA
: 3.38
-6.46
%PPB:
99.89%
LogKaHSA
: 5.52
-3.50
%PPB:
85.22%
LogKaHSA
: 3.75
-3.35
%PPB:
89.25%
LogKaHSA
: 3.87
148
21
Rhynchoph
ylline
22
Hirsutine
23
Biotin
24
cinnamic
acid
25
Galangin
26
Ferulic acid.
27
Sesquiterpe
ne lactone
28
Shogaol
Probability
that
compound
has:
%F(Oral) >
30%: 0.350
%F(Oral) >
70%: 0.060
Probability
that
compound
has:
%F(Oral) >
30%: 0.350
%F(Oral) >
70%: 0.060
Probability
that
compound
has:
%F(Oral) >
30%: 0.944
%F(Oral) >
70%: 0.600
Probability
that
compound
has:
%F(Oral) >
30%: 0.950
%F(Oral) >
70%: 0.838
Probability
that
compound
has:
%F(Oral) >
30%: 0.596
%F(Oral) >
70%: 0.409
Probability
that
compound
has:
%F(Oral) >
30%: 0.950
%F(Oral) >
70%: 0.924
Probability
that
compound
has:
%F(Oral) >
30%: 0.350
%F(Oral) >
70%: 0.207
Probability
that
compound
has:
%F(Oral) >
30%: 0.759
-2.30
%PPB:
77.85%
1.82
L/kg
0.104
0.89
0.89
1.00
0.92
0.68
0.83
2.97
L/kg
0.031
0.37
0.98
0.96
0.87
0.62
0.70
0.26
L/kg
0.005
0.55
0.07
0.38
0.08
0.32
0.39
0.35
L/kg
0.102
0.11
0.12
0.04
0.05
0.07
0.25
0.62
L/kg
0.864
0.37
0.78
0.15
0.45
0.16
0.30
0.34
L/kg
0.057
0.12
0.41
0.01
0.04
0.12
0.38
1.03
L/kg
0.162
0.88
0.87
0.98
0.95
0.87
0.86
2.05
L/kg
0.116
0.26
0.60
0.05
0.12
0.21
0.82
LogKaHSA
: 3.69
-3.14
%PPB:
85.07%
LogKaHSA
: 3.91
-1.90
%PPB:
52.22%
LogKaHSA
: 3.74
-2.36
%PPB:
71.37%
LogKaHSA
: 4.05
-3.47
%PPB:
92.59%
LogKaHSA
: 4.26
-2.02
%PPB:
55.51%
LogKaHSA
: 3.82
-1.43
%PPB:
12.93%
LogKaHSA
: 2.36
4.02
%PPB:
71.11%
LogKaHSA
: 3.98
149
%F(Oral) >
70%: 0.205
29
Neryl
Acetate
30
Indirubin
31
Rotenone
32
Arbutin
Probability
that
compound
has:
%F(Oral) >
30%: 0.350
%F(Oral) >
70%: 0.039
Probability
that
compound
has:
%F(Oral) >
30%: 0.950
%F(Oral) >
70%: 0.773
Probability
that
compound
has:
%F(Oral) >
30%: 0.811
%F(Oral) >
70%: 0.286
Probability
that
compound
has:
%F(Oral) >
30%: 0.060
%F(Oral) >
70%: 0.025
-2.79
%PPB:
93.25%
2.02
L/kg
0.261
0.14
0.72
0.14
0.10
0.06
0.30
1.55
L/kg
0.207
0.32
0.42
0.81
0.35
0.25
0.31
2.07
L/kg
0.454
0.94
0.98
1.00
0.98
0.95
1.00
0.75
L/kg
0.031
0.48
0.33
0.17
0.08
0.03
0.17
LogKaHSA
: 3.55
-4.14
%PPB:
74.27%
LogKaHSA
: 4.04
-4.87
%PPB:
94.84%
LogKaHSA
: 3.67
-1.03
%PPB:
31.05%
LogKaHSA
: 1.85
150
5. Discussion:
5.1Table: 7: structure similarity with commercial drug:
Testolactone (Teslac)
Chrysin
Quercetin
Phytoestrogens
151
Naringenin
Resveratrol
Apigenin
Genistein
Astragalus
152
Emodin
Ellagic Acid
Riboflavin
List of most similar structure with Testolactone (Teslac)
Astragalus
Ellagic Acid
153
Around 170 potential chemo preventive molecules were selected from
pubchem.The docking scores were compared with the reference ligand
score (-17.57kcal/mol).following 32 molecules had lower docking score
than reference molecule.
Diagnosis of Breast cancer is quite difficult. In addition to copying
with potentially life-threatening illness, one must make complex
decision about treatment. Hence its better to prevent cancer than curing
it. At present the best-known options for chemoprevention of Breast
cancer is selective estrogen receptor modulator (SERM)- Testolactone
(Teslac),Anastrozole (Arimidex), Letrozole (Femara), Exemestane
(Aromasin).
This project find out those naturally inhibitor of Breast cancer as better
as Testolactone (Teslac) or better than that. Using various bioinformatics
databases & software we have checked the efficiency of these
chemoprevention molecules & the results suggest that out of 171
molecules. We docked 32 potent Chemo preventive molecules had
lower docking score than the reference molecule. Lower docking score
(more negative) of a molecule the more well it’s effectiveness as an
inhibitor. Thus, sequence which has the least docking score may be an
effective chemo preventive inhibitor of breast cancer.
Now comparing the structure of natural compound & Testolactone
(Teslac).After this comparison I get two structures which are more match with
Testolactone (Teslac) structure. Two natural compounds are Astragalus &
Ellagic Acid.
Conclusion:
Natural molecules include all the small molecules that are
biologically active and those natural molecules which can treat diseases
are under specific trails. Metabolites are also natural molecules which
are used in various treatments like cancer, diabetes, nervous disorders,
etc. metabolites are organic compounds that are not directly involved in
the normal growth, development or reproduction of organisms.
Metabolites among which flavonoids, alkaloids and glycosides were
taken for analysis of their effectiveness against Aromatase for breast
cancer.
Flavonoids, alkaloid and glycosides among the metabolites, were
taken for the docking studies on breast cancer. Testolactone (Teslac) is
one of the successful drugs for breast cancer in spite of its side effects
154
was considered as a reference drug. All the 171 natural molecules were
docked against Aromatase, which is one of the causatives for breast
cancer. Testolactone (Teslac) when docked against 3EQM, gave a
docking score of –(19.6) Testolactone (Teslac) Since is the effective drug,
the molecules showing values close to docking score of Testolactone
(Teslac) were taken as the effective molecules against breast cancer. A
total of top 32 molecules with their docking score were taken. Ascorbic
Acid, Chrysin,
Quercetin, Genistein, Astragalus, Ellagic Acid show the nearest docking
scores to Testolactone (Teslac).So these molecules may possibly prevent
breast cancer.
After comparing the structure of natural compound & Testolactone
(Teslac).After this comparison I get two structures which are more match with
Testolactone (Teslac) structure. Two natural compounds are Astragalus &
Ellagic Acid.
Thus, the appropriate use of a Screening of natural compounds, An
insilico approach on the understanding of its mechanism of action at all
levels, namely at the molecular, cellular, tissue & organ levels as well as
in the animal as a whole without this knowledge we can only make
intuitive decision in selective preventive agent & hope that a useful
chemical result will be forthcoming. More data are needed to accurately
access risk.
TERMINOLOGY:
DRUG --Drug is defined as "a chemical substance used in the treatment,
cure, prevention, or diagnosis of disease or used to otherwise enhance
physical or mental well-being." Drugs may be prescribed for a limited
duration, or on a regular basis for chronic disorders.
DOCKING-- Docking is a method which predicts the preferred
orientation of one molecule to a second when bound to each other to
form a stable complex.Knowledge of the preferred orientation in turn
may be used to predict the strength of association or binding affinity
between two molecules using for example scoring functions.
155
ENERGY MINIMISATION--Energy Minimisation is a non-dynamical
calculation used to find a local potential energy minimum near the
starting structure.Even though molecular dynamics are not being
performed. It is usually needed to do one of these calculations before
attempting MD, because the process of solvating your initial solute
structure usually introduces some bad contacts that need to be relaxed
before you attempt to heat the system, ie. give it kinetic energy.
LIGAND--Ligand (latin ligare = to bind) is a substance that is able to
bind to and form a complex with a biomolecule to serve a biological
purpose. In a narrower sense, it is a signal triggering molecule binding
to a site on a target protein, by intermolecular forces such as ionic
bonds, hydrogen bonds and Van der Waals forces
RECEPTOR--Receptor is a molecule of protein, embedded in a
membrane, to which a mobile signaling (or "signal") molecule may
attach. The membrane may be on the surface of a cell (the "cell
membrane" or "plasma membrane"), or within it, in the cytoplasm
LIPINSKI RULE OF FIVE--Lipinski's Rule of Five is a rule of thumb to
evaluate druglikeness, or determine if achemical compound with a
certain pharmacological or biological activity has propertiesthat would
make it a likely orally active drug in humans. The rule was formulated
by Christopher A. Lipinski in 1997, based on the observation that most
medication drugs are relatively small and lipophilic molecules.The rule
describes molecular properties important for a drug's pharmacokinetics
in the human body, including their absorption, distribution,
metabolism, and excretion ("ADME"). However, the rule does not
predict if a compound is pharmacologically active
Lipinski's Rule of Five states that, in general, an orally active drug has
no more than one violation of the following criteria:
 Not more than 5 hydrogen bond donors (nitrogen or oxygen
atoms with one or more hydrogen atoms)
 Not more than 10 hydrogen bond acceptors (nitrogen or oxygen
atoms)
 A molecular weight under 500 g/mol
 A partition coefficient log P less than 5
156
PARTITION COEFFICIENT(LogP)--The partition coefficient is a ratio
of concentrations of un-ionized compound between the two solutions.
To measure the partition coefficient of ionizable solutes, the pH of the
aqueous phase is adjusted such that the predominant form of the
compound is un-ionized. The logarithm of the ratio of the
concentrations of the un-ionized solute in the solvents is called log P:
AGONIST--An agonist is a substance that binds to a specific receptor
and triggers a response in the cell. It mimics the action of an endogenous
ligand (such as hormone or neurotransmitter) that binds to the same
receptor.
ANTAGONIST--An antagonist acts against and blocks an action
7. References
1. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=644104&loc=ec_rcs
Milewich, L., et al., Ascorbic acid inhibition of aromatase activity in human placenta
tissue. J Steroid Biochem, 1981. 14(2): p. 185-193.
Marsit, J.L., et al., Effects of ascorbic acid on serum cortisol and testosterone:corisol
ratio in junior weightlifters. J Strength and Cond Res, 1998. 12(3): p. 179-184.
• Banhegyi G, Mandl J: The hepatic glycogenoreticular system. Pathol Oncol Res.
2001;7(2):107-10.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B73GT-47F2CVC170&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_sea
rchStrId=1162663291&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVe
rsion=0&_userid=10&md5=0495a60f9858003b07570816cac5aca4
2. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281607&loc=ec_rcs
http://www.tmuscle.com/free_online_article/sports_body_training_performance_bodyb
uilding_supplements/naturally_occurring_aromatase_inhibitors
157
http://www.naturalnews.com/026086_cancer_chrysin_estrogen.html
http://www.vrp.com/articles.aspx?ProdID=art1208&zTYPE=2
3.http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=11821800&loc=ec_rcsA
dlercreutz, H., et al., Inhibition of human aromatase by mammalian lignans and
isoflavonoid phytoestrogens. J Steroid Biochem Mol Biol, 1993. 44(2): p. 147-153.
Chen, S., Y.C. Kao, and C.A. Laughton, Binding characteristics of aromatase inhibitors
and phytoestrogens to human aromatase. J Steroid Biochem Mol Biol, 1997. 61(3-6): p.
107-115
http://www.tmuscle.com/free_online_article/sports_body_training_performance_bodyb
uilding_supplements/naturally_occurring_aromatase_inhibitors
4. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=942&loc=ec_rcs
PDRhealth
Kadohama, N., K. Shintani, and Y. Osawa, Tobacco alkaloid derivatives as
inhibitors of breast cancer aromatase. Cancer Lett, 1993. 75(3): p. 175-182.
Bullion, K., S. Ohnishi, and Y. Osawa, Competitive inhibition of human
placental aromatase
by N-n- octanoylnornicotine and other nornicotine
derivatives. Endocr Res, 1991. 17(3-4): p. 409-419.
Osawa, Y., et al., Aromatase inhibitors in cigarette smoke, tobacco leaves and
other plants. J Enzyme Inhib, 1990. 4(2): p. 187-200
5. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280343&loc=ec_rcs
http://www.drugbank.ca/drugs/DB04216 ………..
Preventive effect of the flavonoid, quercetin, on hepatic cancer in rats via
oxidant/antioxidant activity: molecular and histological evidences.
http://www.raysahelian.com/aromatase.html
6. http://www.drugbank.ca/drugs/DB03467
http://www.ncbi.nlm.nih.gov/sites/entrez
http://www.raysahelian.com/aromatase.html
7. http://www.drugbank.ca/drugs/DB0270
http://www.raysahelian.com/aromatase.html
8. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280443&loc=ec_rcs
http://en.wikipedia.org/wiki/Apigenin
http://www.raysahelian.com/aromatase.html
9. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280961&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01645
http://www.raysahelian.com/aromatase.html
10. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281544&loc=ec_rcs
http://www.naturalnews.com/026514_cancer_brst_cancer_food.html
http://www.raysahelian.com/aromatase.html
11. http://www.drugbank.ca/drugs/DB00668
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=150091&loc=ec_rcs#Prop
erties
http://findarticles.com/p/articles/mi_7396/is_317/ai_n45060728/
http://www.herbalremedies.com/estrogenex.html
158
12http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3220&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=537
13. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=173183&loc=ec_rcs
http://www.freepatentsonline.com/y2004/0156926.html
14. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=73040&loc=ec_rcs
http://pubs.acs.org/doi/abs/10.1021/jm050129p
15. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=22012&loc=ec_rcs
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1241380/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1241194/
16. http://www.quinine-buchler.com/quinicacid.htm. Retrieved September 5 2005
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=1560034&loc=ec_rcs
http://www.drugbank.ca/search/search?query=quinic+acid
http://cancerres.aacrjournals.org/cgi/reprint/42/8_Supplement/3342s.pdf
http://www.freepatentsonline.com/CCL-514-470-p3.html
17. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280805&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01698#
http://www.herbalextractsplus.com/rutin.cfm
http://www.citeulike.org/user/hendryluis/article/5019053
http://www.faqs.org/patents/app/20090053341
18. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280805&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01698#
http://www.herbalextractsplus.com/rutin.cfm
http://www.citeulike.org/user/hendryluis/article/5019053
http://www.faqs.org/patents/app/20090053341
19. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=78151&loc=ec_rcs
http://www.patentstorm.us/patents/4767770/description.html
20. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280899&loc=ec_rcs
http://www.herbalremedies.com/estrogenex.html
http://www.vitamindeal.com/novedex-xt.html
21. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5471851&loc=ec_rcs
http://www.google.co.in/search?hl=en&client=firefoxa&channel=s&rls=org.mozilla:en-US:official&q=poricoic+acid&start=30&sa=N
22. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=68740&loc=ec_rcs
http://www.docguide.com/news/content.nsf/news/852571020057CCF6852573B10077AC
55
23. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280489&loc=ec_rcs
http://www.drugbank.ca/search/search?query=beta-carotene+
http://jnci.oxfordjournals.org/cgi/content/full/98/2/90
24. http://www.drugbank.ca/cgi-bin/show_drug.cgi?CARD=APRD00326
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5479203
http://www.life-enhancement.com/product.asp?ID=80
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
25. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=160512&loc=ec_rcs
http://www.thegoodscentscompany.com/data/rw1384401.html
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
159
26. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=92776&loc=ec_rcs
http://budgetitc.dmsc.moph.go.th/research/pdf/19925.pdf
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1436&Title=Using%20natural%20compounds%20to%20fight%20cancer
http://www.wipo.int/pctdb/ja/ia.jsp?ia=AU2008%2F000908&IA=AU2008000908&DISP
LAY=DESC
27. http://budgetitc.dmsc.moph.go.th/research/pdf/19925.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2758&loc=ec_rcs
http://www.citeulike.org/user/hendryluis/article/5019053
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
28. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6552009&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
http://www.ars-grin.gov/duke/syllabus/module18.htm
http://www.freepatentsonline.com/ACC-514-455.html
29. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=7460&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
30. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=18818&loc=ec_rcs
Beilstein. 5, IV, 451
http://www.ebmonline.org/cgi/rapidpdf/0902-MR-78v1.pdf
31. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=442360&loc=ec_rcs
http://www.wipo.int/pctdb/ja/ia.jsp?ia=AU2008%2F000908&IA=AU2008000908&DISP
LAY=DESC
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
32. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5316215&loc=ec_rcs
http://www.ebmonline.org/cgi/rapidpdf/0902-MR-78v1.pdf
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
33. http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
http://www.drugbank.ca/drugs/DB00169
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280795&loc=ec_rcs
http://clinicaltrials.gov/ct2/show/NCT00867217
http://www.medpagetoday.com/MeetingCoverage/SABCS/17546
http://vmsnews.blogspot.com/2009/12/vitamin-d-reduces-aromatase-inhibitor.html
34. http://www.thedietchannel.com/Is-Essiac-Tea-Helpful-For-Cancer-Treatment.htm
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=65064&loc=ec_rcs
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T6P-4JTXCTN1&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searc
hStrId=1162798882&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVers
ion=0&_userid=10&md5=6ccc1c41f8224ed69ea89c62bdb98de5
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
35. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=14985&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00163Miller ER 3rd, Pastor-Barriuso R, Dalal D,
Riemersma RA, Appel LJ, Guallar E: Meta-analysis: high-dosage vitamin E
supplementation may increase all-cause mortality. Ann Intern Med. 2005 Jan
4;142(1):37-46. Epub 2004 Nov 10.
36. http://eprints.hec.gov.pk/661/1/435.htm
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=11005&loc=ec_rcs
160
http://www.drugbank.ca/drugs/DB02767
http://cancerres.aacrjournals.org/cgi/content/full/66/24/12026?maxtoshow=&HITS=10
&hits=10&RESULTFORMAT=&titleabstract=mushroom&searchid=1&FIRSTINDE
X=0&resourcetype=HWCIT
37. http://www.drugbank.ca/drugs/DB03796
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=985&loc=ec_rcs
http://cancerres.aacrjournals.org/cgi/content/full/66/24/12026?maxtoshow=&HITS=10
&hits=10&RESULTFORMAT=&titleabstract=mushroom&searchid=1&FIRSTINDE
X=0&resourcetype=HWCIT
38. http://www.drugbank.ca/drugs/DB03193
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281&loc=ec_rcs
39. http://www.drugbank.ca/drugs/DB03017
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3893&loc=ec_rcs
http://cancerres.aacrjournals.org/cgi/content/full/66/24/12026?maxtoshow=&HITS=10
&hits=10&RESULTFORMAT=&titleabstract=mushroom&searchid=1&FIRSTINDE
X=0&resourcetype=HWCIT
40. Wolk A, Furuheim M, Vessby B. 2001 Fatty acid composition of adipose tissue and
serum lipids are valid biological markers of dairy fat intake in men. J Nutr
Mar;131(3):828-33.
Smedman AE, Gustafsson IB, Berglund LG, Vessby BO. 1999 Pentadecanoic acid in
serum as a marker for intake of milk fat: relations between intake of milk fat &
metabolic risk factors. Am J Clin Nutr Jan;69(1):22-9.
http://www.lipomics.com/fatty_acids/pentadecanoic-acid
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=13849&loc=ec_rcs
http://pubs.acs.org/doi/abs/10.1021/np000620d
41. http://www.canceractive.com/cancer-active-page-link.aspx?n=525
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=65036&loc=ec_rcs
42, http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5386591&loc=ec_rcs
http://www.foodconsumer.org/newsite/2/Cancer/foods_that_fight_cancer_25102009062
4.html
http://www.canceractive.com/cancer-active-page-link.aspx?n=525&Title=Garlic
43. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=16591&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=525&Title=Garlic
44. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=16590&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=525&Title=Garlic
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
45. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=98280&loc=ec_rcs
http://www.ncbi.nlm.nih.gov/pubmed/9154166
http://www.canceractive.com/cancer-active-page-link.aspx?n=525&Title=Garlic
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
46. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=529&Title=Ellagic%20Acid
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281855&loc=ec_rcs
http://www.physorg.com/news181894265.html
161
47. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6037&loc=ec_rcs
http://www.canceractive.com/cancer-active-pagelink.aspx?n=852&Title=Folic%20Acid
48. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=9064
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T6P-45D13P11&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searc
hStrId=1164188155&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVers
ion=0&_userid=10&md5=9b6087c2d388367984bcd19046c393a2
http://www.ejcancer.info/article/S0959-8049%2804%2900499-X/abstract
http://www.ncbi.nlm.nih.gov/pubmed/12065214
49. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=442988&loc=ec_rcs
http://www.amazing-green-tea.com/green-tea-ingredients.html
http://etd.sun.ac.za/jspui/bitstream/10019/1256/1/Van%20der%20Merwe,%20J%20D.
pdf
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
50. http://www.amazing-green-tea.com/green-tea-ingredients.html
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2519&loc=ec_rcs
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
51http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5429&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01412
http://www.amazing-green-tea.com/green-tea-ingredients.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
52. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
http://www.drugbank.ca/drugs/DB00277
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2153&loc=ec_rcs
http://www.amazing-green-tea.com/green-tea-ingredients.html
53. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=1130&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00152
http://www.amazing-green-tea.com/green-tea-ingredients.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
http://www.ncbi.nlm.nih.gov/pubmed/10215034
54. http://www.drugbank.ca/drugs/DB00140
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=493570&loc=ec_rcs
http://www.amazing-green-tea.com/green-tea-ingredients.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T3G-3VYTKJ0-
162
K&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_sear
chStrId=1164203281&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVer
sion=0&_userid=10&md5=88329c79aba355db8a0f298030360032
55. http://www.amazing-green-tea.com/green-tea-ingredients.html
http://www.drugbank.ca/drugs/DB00165
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=1054&loc=ec_rcs
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
56. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=370&loc=ec_rcs
http://www.amazing-green-tea.com/green-tea-ingredients.html
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1007&Title=Green%20Tea
57. http://www.drugbank.ca/drugs/DB01065
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=896&loc=ec_rcs
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://www.naturalnews.com/022981_melatonin_cancer_brst_cancer.html
58. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3712&loc=ec_rcs
http://www.watchfulwarrior.org/foods.pdf
http://www.canceractive.com/cancer-active-pagelink.aspx?n=1417&Title=Indole%203%20Carbinol%20%20the%20safer,%20natural%20Tamoxifen?
http://www.vrp.com/articles.aspx?ProdID=art1208&zTYPE=2
59. http://www.nutriherb.net/flaxseed.html
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280450&loc=ec_rcs
http://cat.inist.fr/?aModele=afficheN&cpsidt=18378418
60. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280934&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00132
http://www.nutriherb.net/flaxseed.html
http://cat.inist.fr/?aModele=afficheN&cpsidt=18378418
61. http://www.drugbank.ca/drugs/DB04224
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=445639&loc=ec_rcs
http://www.nutriherb.net/flaxseed.html
http://cancerres.aacrjournals.org/cgi/content/full/66/24/12026?maxtoshow=&HITS=10
&hits=10&RESULTFORMAT=&titleabstract=mushroom&searchid=1&FIRSTINDE
X=0&resourcetype=HWCIT
62. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=11128&loc=ec_rcs
http://www.nutriherb.net/flaxseed.html
63. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281303&loc=ec_rcs
http://www.organeem.com/activeconstituentsofneem.html
http://www.canceractive.com/cancer-active-page-link.aspx?n=520&Title=Neem
163
64. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=232706&loc=ec_rcs
http://www.organeem.com/activeconstituentsofneem.html
http://www.canceractive.com/cancer-active-page-link.aspx?n=520&Title=Neem
65. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=200744&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=520&Title=Neem
66. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281310&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=520&Title=Neem
67. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=27439&loc=ec_rcs
http://www.canceractive.com/cancer-active-pagelink.aspx?n=519&Title=Noni%20Juice
68. http://www.citeulike.org/user/hendryluis/article/5019053
http://www.canceractive.com/cancer-active-pagelink.aspx?n=2592&Title=Red%20Clover
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281708&loc=ec_rcs
http://www.medicinehunter.com/redclover.htm
69. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281803&loc=ec_rcs
http://www.medicinehunter.com/redclover.htm
http://www.canceractive.com/cancer-active-pagelink.aspx?n=2592&Title=Red%20Clover
70. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=4133&loc=ec_rcs
http://www.medicinehunter.com/redclover.htm
http://www.canceractive.com/cancer-active-pagelink.aspx?n=2592&Title=Red%20Clover
71. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3314&loc=ec_rcs
http://www.medicinehunter.com/redclover.htm
http://www.canceractive.com/cancer-active-pagelink.aspx?n=2592&Title=Red%20Clover
72. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=104880
http://www3.interscience.wiley.com/journal/109568209/abstract
73.http://www.wipo.int/pctdb/ja/ia.jsp?ia=US2007%2F085385&IA=US2007085385&DI
SPLAY=DESC
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=439242&loc=ec_rcs
http://journals.cambridge.org/action/displayFulltext?type=1&fid=667976&jid=PNS&v
olumeId=41&issueId=01&aid=667968
74. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=91455&loc=ec_rcs
http://journals.cambridge.org/action/displayFulltext?type=1&fid=667976&jid=PNS&v
olumeId=41&issueId=01&aid=667968
http://www.wipo.int/pctdb/ja/ia.jsp?ia=US2007%2F085385&IA=US2007085385&DISP
LAY=DESC
75. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=442089&loc=ec_rcs
http://www.canceractive.com/cancer-active-page-link.aspx?n=516&Title=Stevia
76. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=452967&loc=ec_rcs
164
http://www.canceractive.com/cancer-active-page-link.aspx?n=516&Title=Stevia
77. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=34751
http://www.canceractive.com/cancer-active-pagelink.aspx?n=512&Title=Vitamin%20B-17
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
78. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=4812&loc=ec_rcs
http://www.canceractive.com/cancer-active-pagelink.aspx?n=507&Title=Vitamin%20K
79. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=444899&loc=ec_rcs
https://circle.ubc.ca/handle/2429/2029
80. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5320775&loc=ec_rcs
http://neuro.vetmed.ufl.edu/Alt_med/Library/Monograph/catsclaw.html
http://www.chemspider.com/Search.aspx?q=pteropodine
http://www.jmbblog.com/cat-claw-to-fight-cancer-and-aids/
81. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3033948&loc=ec_rcs
http://www.chemspider.com/Search.aspx?q=pteropodine
http://neuro.vetmed.ufl.edu/Alt_med/Library/Monograph/catsclaw.htm
http://www.jmbblog.com/cat-claw-to-fight-cancer-and-aids/
82. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3037884&loc=ec_rcs
http://neuro.vetmed.ufl.edu/Alt_med/Library/Monograph/catsclaw.html
http://www.jmbblog.com/cat-claw-to-fight-cancer-and-aids/
83. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=120678&loc=ec_rcs
http://neuro.vetmed.ufl.edu/Alt_med/Library/Monograph/catsclaw.html
84. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=171548&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00121
http://www.canceractive.com/cancer-active-page-link.aspx?n=851&Title=Biotin
85. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=444539&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
http://informahealthcare.com/doi/abs/10.3109/14756369009040741?journalCode=enz
86. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5315892&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
http://informahealthcare.com/doi/abs/10.3109/14756369009040741?journalCode=enz
87. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=1183&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
http://www.ebmonline.org/cgi/content/abstract/232/9/1181
88. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=689043&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
http://www.ebmonline.org/cgi/content/abstract/232/9/1181
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
89. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5318691
http://www.draperbee.com/info/propolis.htm
http://www.ebmonline.org/cgi/content/abstract/232/9/1181
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
165
90. http://www.ebmonline.org/cgi/content/abstract/232/9/1181
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=68071&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
91. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281616&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
92. http://www.ebmonline.org/cgi/content/abstract/232/9/1181
http://www.canceractive.com/cancer-active-page-link.aspx?n=2412
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=445858&loc=ec_rcs
http://www.draperbee.com/info/propolis.htm
93. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6036&loc=ec_rcs
http://www.naturalstandard.com/index-abstract.asp?createabstract=/monographs/herbssupplements/chlorella.asp
http://www.breastcancer.org/treatment/hormonal/new_research/20080903b.jsp
http://www.articlesbase.com/health-articles/refined-carbohydrates-the-enemy-ofweight-loss-281071.html/9461024403
94. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=439710&loc=ec_rcs
http://www.naturalstandard.com/index-abstract.asp?createabstract=/monographs/herbssupplements/chlorella.asp
http://www.drugbank.ca/drugs/DB01869
http://www.vrp.com/pdf/mayNews2004.pdf
95. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3884&loc=ec_rcs
http://health.centreforce.com/health/graviola_brochure.html
http://sify.com/news/cancer-patients-should-beware-supplement-use-news-healthjgma4cceiga.html
http://www.wipo.int/pctdb/ja/ia.jsp?ia=US2008%2F005656&IA=US2008005656&DISP
LAY=DESC
96. http://sify.com/news/cancer-patients-should-beware-supplement-use-news-healthjgma4cceiga.html
http://www.wipo.int/pctdb/ja/ia.jsp?ia=US2008%2F005656&IA=US2008005656&DISP
LAY=DESC
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=72734&loc=ec_rcs
http://www.herbs2000.com/herbs/herbs_pau_darco.htm
97. http://jpet.aspetjournals.org/content/297/3/1099.full
http://www.healthkey.com/hw-hn-2172008,0,5669573.healthwisestory
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=338659&loc=ec_rcs
http://en.wikipedia.org/wiki/Sesquiterpene_lactone
98. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=240&loc=ec_rcs
http://wwwchem.uwimona.edu.jm/lectures/guava.html
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TF8-4PYJS8FC&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searc
hStrId=1167121897&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVers
ion=0&_userid=10&md5=b621507f3a33f82dc6ccdeaafbac3bae
99. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=19725&loc=ec_rcs
http://www.pherobase.com/database/compound/compounds-detail-alpha-copaene.php
166
http://www.naturalstandard.com/index-abstract.asp?createabstract=/monographs/herbssupplements/cassia.asp
100. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=31253&loc=ec_rcs
http://health.centreforce.com/health/graviola_brochure.html
http://www.ema.europa.eu/pdfs/human/hmpc/humulus_lupulus/51361806en.pdf
101. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8908&loc=ec_rcs
http://wwwchem.uwimona.edu.jm/lectures/guava.html
http://www.wipo.int/pctdb/ja/wo.jsp?WO=2006041939&IA=US2005035861&DISPLAY
=DESC
102. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8048&loc=ec_rcs
http://wwwchem.uwimona.edu.jm/lectures/guava.html
http://www.wipo.int/pctdb/ja/wo.jsp?WO=2006041939&IA=US2005035861&DISPLAY
=DESC
103. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281520&loc=ec_rcs
http://wwwchem.uwimona.edu.jm/lectures/guava.html
http://www.wipo.int/pctdb/ja/wo.jsp?WO=2006041939&IA=US2005035861&DISPLAY
=DESC
104. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=4835
http://www.springerlink.com/content/p34l573502002287/
http://www.dickcontino.com/beet-root.htm
105. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=68406&loc=ec_rcs
http://www.ncbi.nlm.nih.gov/pubmed/17238068
106.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=3033866&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://books.google.co.in/books?id=g_OCl6P0vsYC&pg=PA58&lpg=PA58&dq=Bisabol
ene+%2Baromatase+inhibitor&source=bl&ots=audEFALPs&sig=hQ3i2Z5ak3B68XE4963YH_za5no&hl=en&ei=jvZPS1HgebsA_PG7aMK&sa=X&oi=book_result&ct=result&resnum=1&ved=0CAkQ6AEw
AA#v=onepage&q=Bisabolene%20%2Baromatase%20inhibitor&f=false
107. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=7460&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
108. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=638011&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
109. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8842&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
110. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
167
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6549&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
111. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=22311&loc=ec_rcs
http://www.friedli.com/herbs/ginger.htm
112. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6455496&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
113. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6616&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
114. http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=90942&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
115. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281794&loc=ec_rcs
http://www.friedli.com/herbs/ginger.html
http://breastcancerisabitch.blogspot.com/2009/02/foods-that-fight-cancer.html
http://humupd.oxfordjournals.org/cgi/reprint/13/5/487.pdf
116. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281224&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
117. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280435&loc=ec_rcs
http://www3.interscience.wiley.com/journal/114300439/abstract?CRETRY=1&SRETR
Y=0
118. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281703&loc=ec_rcs
http://www3.interscience.wiley.com/journal/114300439/abstract?CRETRY=1&SRETR
Y=0
119. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5280445&loc=ec_rcs
http://www3.interscience.wiley.com/journal/114300439/abstract?CRETRY=1&SRETR
Y=0
120. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281628&loc=ec_rcs
http://www3.interscience.wiley.com/journal/114300439/abstract?CRETRY=1&SRETR
Y=0
121.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=24199357&loc=ec_rcs
http://www.wholeworldbotanicals.com/herbal_graviola
122. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5321483&loc=ec_rcs
http://www.wholeworldbotanicals.com/herbal_graviola
123.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?sid=75289216&loc=es_rss
http://www.wholeworldbotanicals.com/herbal_graviola
124. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=178503&loc=ec_rcs
http://www.wholeworldbotanicals.com/herbal_graviola
168
125. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5321489&loc=ec_rcs
http://www.wholeworldbotanicals.com/herbal_graviola
126. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=312871&loc=ec_rcs
http://www.wholeworldbotanicals.com/herbal_graviola
127. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6537493&loc=ec_rcs
http://www.ispub.com/journal/the_internet_journal_of_third_world_medicine/volume_
8_number_2_16/article/assessment-of-the-antibacterial-activity-of-vernoniaamygdalina-and-occimum-gratissimum-leaves-on-selected-food-borne-pathogens.html
128. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=323&loc=ec_rcs
http://www.ispub.com/journal/the_internet_journal_of_third_world_medicine/volume_
8_number_2_16/article/assessment-of-the-antibacterial-activity-of-vernoniaamygdalina-and-occimum-gratissimum-leaves-on-selected-food-borne-pathogens.html
129. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?sid=11767
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T0P-47XN7JCH&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_sear
chStrId=1158395478&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVer
sion=0&_userid=10&md5=0cb3522f6aa069b7d886acf6dad29033
130. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6436299&loc=ec_rcs
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T0P-47XN7JCH&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_sear
chStrId=1158395478&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVer
sion=0&_userid=10&md5=0cb3522f6aa069b7d886acf6dad29033
131. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5281472&loc=ec_rcs
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T0P-47XN7JCH&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_sear
chStrId=1158395478&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVer
sion=0&_userid=10&md5=0cb3522f6aa069b7d886acf6dad29033
132. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=148189
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://www.springerlink.com/content/t6vqp6u0ta6gawjn/
133. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6442554&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://science.thomsonreuters.com/m/pdfs/mgr/ddf_thesaurus3.pdf
134. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=440968&loc=ec_rcs
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://dietmedics.com/pages/qck_main.html?c=k&s=
135. http://www.chemspider.com/Chemical-Structure.2265.html
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://www.technology-catalysts.com/pdf/nut4bro.pdf
136. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=17100&loc=ec_rcs
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://www.technology-catalysts.com/pdf/nut4bro.pdf
169
137. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8294&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
138. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=637566&loc=ec_rcs
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://www.technology-catalysts.com/pdf/nut4bro.pdf
139. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=1549026&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://dietmedics.com/pages/qck_main.html?c=k&s=
140. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=403919&loc=ec_rcs
http://www.organicfacts.net/health-benefits/essential-oils/health-benefits-of-bergamotessential-oil.html
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://dietmedics.com/pages/qck_main.html?c=k&s=
141. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=64971&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
142.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=16132398&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
143. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2909&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00091
http://jpet.aspetjournals.org/content/293/1/230.full
144. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=4192&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00683
http://jpet.aspetjournals.org/content/293/1/230.full
145. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5556&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00897
http://jpet.aspetjournals.org/content/293/1/230.full
146. http://www.drugbank.ca/drugs/DB00342
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5405&loc=ec_rcs
http://jpet.aspetjournals.org/content/293/1/230.full
147. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=60787&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01232
http://jpet.aspetjournals.org/content/293/1/230.full
148. http://www.drugbank.ca/drugs/DB00977
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5991&loc=ec_rcs
http://jpet.aspetjournals.org/content/293/1/230.full
149. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6475943&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://www.medworm.com/rss/search.php?qu=%22breast+cancer%22&t=Breast%20C
ancer&f=cancer&s=Search&r=Any&ftc=13&page=7&o=d
170
150. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5359405&loc=ec_rcs
http://www.wipo.int/pctdb/ja/ia.jsp?ia=US2008%2F053798&IA=US2008053798&DISP
LAY=CLAIMS
151. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=265237&loc=ec_rcs
http://www.technology-catalysts.com/pdf/nut4bro.pdf
http://mct.aacrjournals.org/content/early/recent
152. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5288826&loc=ec_rcs
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.drugbank.ca/drugs/DB00295
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
153. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5284371&loc=ec_rcs
http://www.drugbank.ca/drugs/DB00318
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
154. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6167
http://www.ikisan.com/mp/cache/a_introduction.asp .
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
155. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=10219&loc=ec_rcs
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
156. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=32681&loc=ec_rcs
http://www.drugbank.ca/drugs/DB01180
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
157. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=20367&loc=ec_rcs
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
158. http://www.drugbank.ca/drugs/DB00570
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8935
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
159. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5978&loc=ec_rcs
http://www.ikisan.com/mp/cache/a_introduction.asp
http://www.articlesbase.com/health-articles/aromatherapy-massage-oils-to-use-andrelax-1425694.html
160. http://augmentinforce.50webs.com/ANTI%20ESTROGEN%20SOLUTIONS.htm
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2758&loc=ec_rcs
http://ezinearticles.com/?Basil-Oil-%28Ocimum-Basilicum%29---What-It-Is-And-
171
Why-To-Use-It&id=655944
http://www2.sbc.edu/marcus/2004conf_abstracts.html
161. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2537&loc=ec_rcs
http://ezinearticles.com/?Basil-Oil-%28Ocimum-Basilicum%29---What-It-Is-AndWhy-To-Use-It&id=655944
http://augmentinforce.50webs.com/ANTI%20ESTROGEN%20SOLUTIONS.htm
http://www2.sbc.edu/marcus/2004conf_abstracts.html
162. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=5463455&loc=ec_rcs
http://ezinearticles.com/?Basil-Oil-%28Ocimum-Basilicum%29---What-It-Is-AndWhy-To-Use-It&id=655944
http://augmentinforce.50webs.com/ANTI%20ESTROGEN%20SOLUTIONS.htm
http://www2.sbc.edu/marcus/2004conf_abstracts.html
163. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8815&loc=ec_rcs
http://ezinearticles.com/?Basil-Oil-%28Ocimum-Basilicum%29---What-It-Is-AndWhy-To-Use-It&id=655944
http://augmentinforce.50webs.com/ANTI%20ESTROGEN%20SOLUTIONS.htm
http://www2.sbc.edu/marcus/2004conf_abstracts.html
164. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=637563&loc=ec_rcs
http://www.holistic-online.com/Herbal-med/_Herbs/h55.htm
http://www3.interscience.wiley.com/journal/41923/abstract
165. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=445070&loc=ec_rcs
http://derrida.en.ecplaza.net/214.asp
http://www.jenniferhaas.com/aromatherapy.htm#02usages
166. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=11552&loc=ec_rcs
http://derrida.en.ecplaza.net/214.asp
http://www.jenniferhaas.com/aromatherapy.htm#02usages
167. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8175&loc=ec_rcs
http://derrida.en.ecplaza.net/214.asp
http://www.jenniferhaas.com/aromatherapy.htm#02usages
168. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=92987&loc=ec_rcs
http://chestofbooks.com/health/materia-medica-drugs/PharmacologyTherapeutics/Granatum-Granati-Pomegranate-U-S-P.html
169. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=6758&loc=ec_rcs
http://toxsci.oxfordjournals.org/cgi/reprint/82/1/70.pdf
170. http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=440936&loc=ec_rcs
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T8D-4TJ1HKH1&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searc
hStrId=1167173779&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVers
ion=0&_userid=10&md5=1cf4890b5ce3187792e795ab0086ed37
http://skincare-cosmeceuticals.com/formulations/index.php?topic=18.0
171. http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T8D-4TJ1HKH1&_
user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_searchStr
I
d=1167173779&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVersion=0
172
&_userid=10&md5=1cf4890b5ce3187792e795ab0086ed37
http://skincare-cosmeceuticals.com/formulations/index.php?topic=18.0
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=68171&loc=ec_rcs