Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
CA 02538494 2012-07-25
COMPOSITION FOR MODULATING BLOOD PARAMETERS
FIELD OF THE INVENTION
This invention relates to the field of nutritional supplements.
BACKGROUND OF THE INVENTION
Phytosterols are fats that are present in all plants, including fruits and
vegetables. Although structurally similar to cholesterol, the sterols
synthesized by
animals and plants differ in the nature of their side chain (see Figure 1)
(Allayee et al.
2000, Science 290: 1709-1711).
In animals, cholesterol is the most abundant sterol. In plants, more than 40
sterols have been identified, of which beta-sitosterol, stigmasterol, and
campesterol
are the most common (Hicks and Moreau 2001, Food Technol 55: 63-67). These
phytosterols occur in free form or are esterified to free fatty acids, sugar
moieties or
phenolic acids (Baker et al. 1999, Food Chem Toxicol 37: 13-22). Studies have
demonstrated that beta-sitosterol possesses anti-inflammatory and immune
modulating properties. It has been estimated that consumption of 3 g/day of
phytosterols could reduce the risk of heart disease by 15% to 40%, depending
on age
and other dietary factors (Hicks and Moreau 2001). However, the western diet
typically contains 100-300mg of plant sterols (Nguyen 1999, J Nutr 129: 2109-
2112).
EnzogenolTM is an example of a water soluble extract of monomeric and
oligomeric proanthocyanidins, flavonoids, flavonoid glycosides, esters and
natural
organic acids prepared from the bark of Pinus radiate by a pure water
extraction
process (Shand et al. 2003, Phytother Res 17: 490-494). The extract has been
shown
to have in vitro antioxidant action as measured by inhibition of micelle
oxidation, red
blood cell hemolysis and a nitro blue tetra zolium enzymatic method (Gieseg
and
Baird 1998, Free Rad Biol Med 25: S104; Wood et al. 2002, Food Chem 77: 155-
161).
CA 02538494 2012-05-23
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Omega 3, 6 and 9 fatty acids, such as found in Cellasate, have a structural
role
in phospholipids of all cell membranes in the body, influencing membrane
viscosity
and permeability (Drevon 1992, Nutr Rev 50: 38-45). Eicosapentaenoic acid
(EPA)
and docosahexaenoic acid (DHA) are thought to be responsible for beneficial
effects,
such as prevention and management of coronary heart disease and hypertension
(Simopoulos 1999, Am J Clin Nutr 70: 560S-569S).
SUMMARY OF THE INVENTION
According to a first aspect of the invention, there is provided the use of an
effective amount of a composition comprising:
10% to 90% phytosterols;
5% to 85% essential fatty acid complexes; and
5% to 85% proanthocyanidins, flavonoids and polyphenols for modifying blood
lipid parameters in an individual.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
Unless defined otherwise, all technical and scientific terms used herein have
the same meaning as commonly understood by one of ordinary skill in the art to
which
the invention belongs. Although any methods and materials similar or
equivalent to
those described herein can be used in the practice or testing of the present
invention,
the preferred methods and materials are now described. All publications
mentioned
hereunder are incorporated herein by reference.
As used herein, the term "treating" in its various grammatical forms refers to
curing, reversing, attenuating, alleviating, minimizing, suppressing or
halting the
deleterious effects of a disease state, disease progression, disease causitive
agent or
other abnormal condition.
Described herein is a new composition or nutritional supplement composed of
natural products. In some embodiments, the supplement may be used by
individuals
desirous of regulating their immune 'system, for example, to prevent and treat
diseases thereof. As discussed below, the composition combines phytosterols
with
anti-oxidants and fatty acid complexes.
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The composition has also been demonstrated to be beneficial in the
modification of blood lipid parameters, as discussed below. Specifically, the
composition or supplement has been shown to reduce the total cholesterol (TC)
to
high density lipoprotein (HDL) ratio (TC/HDL) as well as to reduce the low
density
lipoprotein (LDL) to HDL ratio (LDUHDL) compared to a placebo-treated control,
as
discussed below. As will be appreciated by one of skill in the art, the
composition or
supplement at formulae and/or dosages described herein is in some embodiments
of
the invention administered to an individual in need of such treatment or used
for
modifying the blood lipid parameters of said individual, for example, reducing
the
TC/HDL ratio or reducing the LDUHDL ratio. As will be appreciated by one of
skill in
the art, examples of such individuals include but are by no means limited to
individuals having, at risk of having or suspected of having
hypercholesterolemia or
otherwise having or suspected of having elevated cholesterol levels or reduced
HOL
levels.
As discussed below, administration of the composition or supplement to an
individual in need of such treatment reduced basophil and IL-6 levels in said
individual
compared to a second individual administered a placebo control.
In another embodiment, administration of the supplement as described herein
to an individual in need of such treatment reduced LDL levels, increased HDL
levels,
reduced the LDUHDL ratio or reduced the total cholesterol to HDL ratio in said
individual compared to a second individual administered a placebo control.
In one embodiment, the composition or supplement comprises: 10% to 90%
phytosterols, 5% to 85% essential fatty acid complexes and 5-85% antioxidants.
In
other embodiments, the supplement comprises 20% to 90% phytosterols, 5% to 75%
essential fatty acid complexes and 5-75% antioxidants. In another embodiment,
the
composition or supplement comprises: 10% to 80% phytosterots, 10% to 80%
essential fatty acid complexes and 10-80% antioxidants. In other embodiments,
the
supplement comprises 20% to 80% phytosterols, 10% to 80% essential fatty acid
complexes and 10 to 80% antioxidants. In a preferred embodiment, the
supplement
comprises 29% to 75% phytosterols; 10% to 50% essential fatty acid complexes;
and
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5% to 50% antioxidants. The supplement or composition may be arranged In the
form
of a capsule or other suitable, ingestible format, such as a tablet, pill or
other similar
form known in the art.
In a preferred embodiment, the supplement or composition comprises 29% to
75% beta-sitosterol; 10% to 50% essential fatty acid complexes; and 5% to 50%
antioxidants.
In a yet further preferred embodiment, there is provided a single-dose
supplement or composition comprising: 300 mg phytosterols; 50 mg essential
fatty
acid complexes; 20 mg antioxidants; and 30 mg micro-cellulose filler. As
discussed
herein, this composition may be taken once daily or twice daily.
In some embodiments, the above-described compositions are used for treating,
preventing or ameliorating symptoms associated with an immune system disease.
In
these embodiments, the composition or supplement at a dosage, concentration or
range described above is administered to an individual in need of such
treatment.
Examples of immune system diseases include but are by no means limited to
colds,
flu, diabetes, allergies, asthma, pneumonia, fibromyalgia, HIV infection,
hepatitis C
infection, and multiple sclerosis. In a preferred embodiment, the composition
at
dosages, concentrations or ranges described above is used to treat or prevent
pneumonia, fibromyalgia or a hepatitis C infection. In a yet preferred
embodiment, the
composition or supplement described above is used to treat an immune system
disease as described above by administering the composition at a concentration
or
dosage as described to an individual in need of such treatment.
Accordingly, in yet another embodiment, there is provided a method of
decreasing the LDUHDL ratio or reducing the TC/HDL ratio in an individual in
need of
such treatment comprising administering to said individual an effective amount
of a
supplement comprising 10% to 90% phytosterols, 5% to 85% essential fatty acid
complexes and 5-85% antioxidants. In other embodiments, the supplement
comprises
20% to 90% phytosterols, 5% to 75% essential fatty acid complexes and 5-75%
antioxidants. In another embodiment, the composition or supplement comprises:
10%
to 80% phytosterols, 10% to 80% essential fatty acid complexes and 10-80%.
CA 02538494 2006-02-24
antioxidants. In other embodiments, the supplement comprises 20% to 80%
phytosterols, 10% to 80% essential fatty acid complexes and 10 to 80%
antioxidants.
In a preferred embodiment, the supplement comprises 29% to 75% phytosterols;
10%
to 50% essential fatty acid complexes; and 5% to 50% antioxidants. In a
further
5 preferred embodiment, the supplement or composition comprises 29% to 75%
beta-sitosterol; 10% to 50% essential fatty acid complexes; and 5% to 50%
antioxidants. In a yet further preferred embodiment, there is provided a
single-dose
supplement or composition comprising: 300 mg phytosterols; 50 mg essential
fatty
acid complexes; 20 mg antioxidants; and 30 mg micro-cellulose filler. As
discussed
herein, this composition may be taken once daily or twice daily.
The inherent ingenuity of the invention Is the discovery of the range of the
amount of beta-sitosterol that is the most effective to achieve the goal of
regulating
the Human Immune System and the prevention and/or treatment of diseases
thereof.
As a result, this new composition need only be taken once a day to maintain
general
health.
The further uniqueness of the invention is the discovery that combining
phytosterols with anti-oxidants and fatty acid complexes produces a
synergistic effect
whereby the cumulative total effect of the composition is greater than the sum
of the
individual effects of the respective components, As a result, this new
composition
optimizes the immune system to deal with viral and bacterial infections, while
also
protecting the body from degeneration due to free radical damage.
Additionally, the invention is original and novel because it can not only
assist in
preventing possible causes of cancer, but, it inhibits or reduces cancer cell
growth in
breast, colon and prostate cancer cases, and facilitates the removal of the
dead tissue
arising from the death of the said tumors. That is, the composition can
modulate or
optimize, as opposed to only boosting, the immune system, as discussed below.
The utility of said invention is further demonstrated and confirmed by the
fact
that it accomplishes its goals with none of the physical or psychological side
effects of
traditional treatments such as radiation, chemotherapy and surgery.
The basis of the present invention is the use of a combination of
phytosterols,
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anti-oxidants and fatty acid complexes for the following purposes:
1, To modulate the Human Immune System;
2. To prevent or treat diseases of the Human Immune System, for example
but by no means limited to:
Common cold and flu;
Microorganism infections, for example viral, yeast and bacterial infections;
Allergies;
Rheumatoid Arthritis;
Lupus;
Chronic Fatigue Syndrome;
Fibromyalgia;
Asthma;
Diabetes; and
Eczema.
3. To inhibit or reduce cancer cell growth in breast, colon and prostate
cancer cases;
4. To maintain CD4 Lymphocyte counts in HIV patients;
5. To treat enlarged prostate problems;
(5, As an anti-inflammatory agent; and
7. Modifying blood lipid parameters, as discussed below.
As discussed above, the compositions described herein act to modulate the
immune system. Specifically, the compositions act to restore normal functions
in an
individual in need thereof. As will be apparent to one of skill in the art,
this explains
how the compositions are able to treat diseases or conditions characterized by
immune activation, such as autoimmune disorders such as arthritis and
inflammation,
as well as diseases characterized by immune suppression or deficiency, such as
acquired immune deficiency syndrome. While not wishing to be bound to a
specific
theory, the inventors believe that the composition acts to modulate immune
function
such that an immune system that is acting abnormally, that is, is either
depressed or
chronically activated, returns to normal functioning, that is, returns to the
state of
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activation found in most individuals who are not currently undergoing a
challenge to
their immune system.
Although preferred compositions to achieve these ends are listed below, they
are by no means to be interpreted as limiting the broad scope of the present
invention
as various other combinations of phytosterols, anti-oxidants and fatty acid
complexes
may achieve the same beneficial results.
The preferred composition of the present application consists of 400mg
capsules containing 300mg of phytosterols (a minimum of which is 117mg of
beta-sitosterol), 50mg essential fatty acid complexes, 20mg broad spectrum
antioxidants, and 30mg micro-cellulose filler. As will be appreciated by one
of skill in
the art, this represents an example of an effective amount of the composition.
Other
suitable concentrations and dosages may be determined using the ranges
described
herein. As will be apparent to one of skill in the art, as used herein,
'effective amount"
refers to an amount that achieves the desired effect.
In another preferred form, the composition comprises from 29% to 75%
beta-sitosterol, 10% to 50% essential fatty acid complexes and from 5% to 50%
antioxidants. As will be appreciated by one of skill in the art, other
suitable,
pharmaceutically acceptable fillers may also be used in the invention.
In other embodiments, there is provided a composition comprising: 1-95%
phytosterols; 1-95% essential fatty acid complexes; and 1-95% antioxidants.
The
composition may further include 1-95% filler, for example, micro-cellulose
filler. In
other embodiments, 5-05% of the phytosterols may be beta-sitosterol.
A further innovative feature of the invention is the discovery that the
essential
fatty acid complexes together are not only themselves an immune system
modulator,
but also facilitates the body's absorption of beta-sitosterol and
antioxidants.
This invention is a composition of natural products that is designed to
stimulate
the body to marshal its own natural defenses against allergies and other
immune
related disorders, breast, colon and prostate cancer, and AIDS, as opposed to
alleviating or preventing these disorders with chemical or synthetic products.
Many of
the traditional treatments have severe side effects, which in the case of
prostate
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cancer can include impotence and incontinence, and in the case of
chemotherapy,
severe depression of the immune system. Traditional treatments also only treat
the
tumors, i.e. the symptoms, and not the underlying cause, and in the case of
chemotherapy, leave dead tissue to be removed from the body.
One of the suspected causes of cancer is free radical damage on cellular
components such as DNA or the cell membranes. Free radicals are the natural
by-products of many processes within and among cells. They are also created by
exposure to various environmental factors such as tobacco smoke and radiation.
Once formed these highly reactive radicals can start a chain of reactions that
can
result in damage to cell walls, certain cell structures, and the genetic
material within
the cells such as DNA.
In the preferred form this composition contains one of the world's most
advanced anti-oxidant complexes, EnzogenolT"", produced by Enzo Nutraceuticals
through a patented process that creates an aqueous extract from Pinus Radiata
(pine
bark), which is a superior source of proanthocyanidins and fiavonoids. This
technically
advanced anti-oxidant complex provides an excellent source of free radical
scavengers:
In addition to scavenging free radicals, the EnzogenoJT"" in this composition
can
stimulate phagocytosis, an immune system response in which cells ingest and
eliminate harmful micro-organisms and foreign heavy metals, and hence
facilitates the
removal of the by-products of cell death.
Research has shown that the Human Immune System can be stimulated by
beta-sitosterol, This composition contains this remarkable phytonutrient which
has
been shown to be effective In Inhibiting cancer cell growth and accelerating
cancer
cell death in breast, prostate and colon cancer. It can also reduce metastases
to
lymph nodes.
Research at the University of New York at Buffalo by Awad, Downey and Fink
has shown that beta-sitosterol can inhibit breast cancer cell growth by 66%
after 3
days and 80% after 5 days. It can also increase the rate of cell death by 600%
(Int. J.
Mol. Med 2000 May 5(5)). Research by the same group has also shown that
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beta-sitosterol can inhibit prostate cancer cell growth by 24% and increase
cell death
by 400% (Nutr Cancer 1998; 32 (1)). Similar research on colon cancer cells
showed a
reduction In cell growth of 55% (Anti Cancer Res, 1998 Mar-Apr 18 (2a)). The
exact
mechanism behind the inhibition of cancer cell growth and the increase in
apoptosis is
not yet fully understood, but may be related to the increase in T-cell
proliferation
produced by beta-sitosterol (Int. JnI. Immuno Pharmacol 1996 vol 18 Bouic at
al).
Beta-sitosterol has been shown to maintain CD4 lymphocyte count in AIDS
patients
(AIDS Bulletin Summer 97).
The potential benefits of the antioxidants together with the beta-sitosterol
to
scavenge free radicals and to kill and remove tumors can only be realized if
the
human body actually absorbs both the anti-oxidant complex and the
betasitosterol.
The role of the essential fatty acid complex is to ensure that the antioxidant
complex
and the beta-sitosterol are absorbed. The essential fatty acid complex itself
is a blend
of essential fatty acids, amino acids, praline rich peptides, short chain
polypeptides
and enzymes. This complex also happens to be an immune system modulator, and
consequently also supports the body's attack on cancer by promoting and
supporting
the T-cell proliferation initiated by the beta-sitosterol.
The combined effects of the antioxidant complex, the beta-sitosterol and the
essential fatty acid complex on breast, colon and prostate cancers are to
reduce the
risk of the formation of new cancers through the more effective scavenging of
free
radicals, to inhibit the growth and increase the rate of cell death of the
tumors, and to
assist the body in disposing of dead tumor cells.
This formulation has been used to treat a 68 year old male with a PSA of 17.
Over a six month period his PSA, (the normal measure of prostate cancer which
would normally increase over time), dropped to 11.8, a reduction of over 30%,
with no
side effects.
The combined immune modulating properties of beta-sitosterol, antioxidant
complex, and the essential fatty acid complex also make this formulation an
excellent
anti-allergen with no side effects which Is also new and unique in the field
of
anti-allergens. This formulation has been used to treat several adults with
ages
CA 02538494 2006-02-24
ranging between 22 and 52 with excellent results occurring in the space of
five to six
days, again with none of the side effects normally associated with anti-
allergens.
Early trials of this unique formulation have also shown the capacity to
alleviate
such immune related disorders as rheumatoid arthritis, again with none of the
adverse
5 side effects normally associated with the remedies traditionally used in
this area.
In addition, the effectiveness of beta-sitosterol in maintaining the CD4
lymphocyte count in AIDS patients can be increased by the addition of the
antioxidant
and essential fatty acid complexes in this formulation, again providing an
important
new application for this new and unique formulation.
10 Normally capsules are taken once a day, in a preferred embodiment, at least
one hour or at least half an hour before and at least 2 hours after food, with
water or
fruit juice (NOT milk or any other fat containing liquid). As will be
appreciated by one
of skill the art, these time frames and frequencies are suggested for most
users, but
other users may find best results taking the capsule more frequently or less
frequently, or, for example, with meals or closer to meals.
The term "phytosterols" is to be construed to include, but not to be
restricted to,
beta-sitosteroi, atigmasterol, campesterol, brassicasterol together with their
associated glucosides.
The term "antioxidant complex" is to be construed to include, but not to be
restricted to, a broad range of proanthocyanidins, flavonoids and polyphenols.
The term "essential fatty acid complex" is to be construed to include, but not
restricted to, amino acids, essential fatty acids, peptides, proline rich poly
peptides,
and various digestive enzymes.
The following are examples of usages, showing the utility of the composition.
Example I - Allergies
The composition was used by an individual to treat seasonal allergies. The
individual found that one or more of the symptoms associated with allergies,
including
runny nose, red itchy eyes, aching joints and a feeling of always being tired,
were
ameliorated following taking the composition. Specifically, users found that
the
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symptoms were ameliorated after 24 hours and remained symptom free after 2
weeks
by taking one dosage or capsule per day.
Another individual who suffered from moderate asthma and allergies to dust
and animal hair found that taking the composition relieved or ameliorated
symptoms
including itchy and watery eyes and a runny nose. This individual also
reported
increased energy and an improved sense of well-being.
Another individual who suffered from allergies to dust mites which led to a
chronically stuffed nose that would drip constantly found that these symptoms
were
ameliorated following taking the composition over an initial regimen and then
taking
the composition daily on an as needed basis.
Another individual reported allergies to pollen and other seasonal allergies
and
found that taking the composition over a period of a few weeks ameliorated the
allergy-related symptoms, including itchy eyes and sneezing.
Example 2 - ASTHMA
An individual who suffered from asthma found that following taking the
composition regularly for 3 months, the asthma-associated symptoms were
ameliorated.
Another individual who suffered from asthma and food allergies and had
symptoms including itchy eyes and a runny nose found that within 2 weeks of
taking
the composition on a regular basis, she was off the antihistamines and her
stuffy nose
was completely gone. Furthermore, she found that her asthma progressively
improved and was able to eat foods that she couldn't eat before without having
immediate allergic reactions.
Example 3. COLDS & FLU
One individual who had contracted pneumonia took two capsules of the
composition per day over a five day period and found that the symptoms
associated
with pneumonia were ameliorated,
Another individual who also had a cold develop into pneumonia found that the
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symptoms associated with the illness were ameliorated during a two month
regimen
of taking the composition daily.
Another individual took the composition regularly to prevent colds or "flu".
This
individual found less frequent occurrences of colds and "flu" and also found
that taking
the composition on a regular basis provided "an all over feeling of
wellbeing".
Example 4 - DIABETES
A diabetic who was using insulin 3 times per day for the last 12 years, at an
average of 50 units a day pf insulin, consisting of 20 units in the morning,
20 units in
the afternoon and 10 units before bed found that after having taken the
composition
daily for 3 weeks that the individual was able to eliminate the night-time
units.
Example 5 - ECZEMA
An individual who suffered from eczema, particularly under her armpits on both
sides, would also get eczema on her hands whenever her hands were exposed to
any
chemicals or detergents (ie doing any cleaning) and even after folding the
laundry.
She managed the eczema with cortisone cream-a few times per week. After 2
months
of taking the composition along with eliminating certain food allergies, her
eczema
was 90% gone (in places where it had been chronic for years) and her hands
(which
used to be effected almost constantly) were completely clear,
Example 6. FIBROMYALGIA
An individual who suffered from fibromyalgia found that taking one dose of the
composition one hour before breakfast helped ameliorate symptoms including
pain
and insomnia.
Another individual with fibromyalgia reported wakening 3-4 times per night
with
severe cramps in their feet and calves. Following taking the composition on a
regular
dosage regimen, this individual found amelioration of many of the symptoms
associated with the fibromyalgia.
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Example 7 - HEPATITIS C
An individual who suffers from hepatitis C reported a progressive increase in
energy during the first 2 weeks of taking the composition.
Example 8 - HIV
An individual having an HIV infection reported that their CD-4 level climbed
by
300 points following a 3 month regimen of taking the composition.
Example 9 - MULTIPLE SCLEROSIS
An individual with multiple sclerosis reported increased energy and improved
balance on taking the composition.
Example 10 - URINARY FLOW
An individual suffering from prostate enlargement reported that following
taking
the composition, the time between urinations was increased and also reported
increased flow.
Example 11 - PELVIC INFLAMMATORY DISEASE
Several users have reported that the composition as described above
alleviated one or more of the symptoms associated with pelvic inflammatory
disease.
Example 12 - RHEUMATOID ARTHRITIS
An individual with rheumatoid arthritis reported that following taking two
capsules of the composition per day for two weeks, many of the symptoms
associated
with the rheumatoid arthritis were ameliorated.
Dosage arxd Administration
The recommended adult dosing regimen is currently one capsule per day by
oral ingestion. In most embodiments, the supplement is taken with water or
juice (not
milk), on an empty stomach 45 minutes before eating. One capsule a day
normally
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maintains general health. More serious conditions may require two capsules per
day,
one in the morning and one at night. The dosing regimen can be changed, for
example, to two capsules per day for the first week, followed by one capsule
per day.
Pharmacology
Phytosterols:
Beta-sitosterol is not synthesized endogenously in mammals. Animal studies
have demonstrated its intestinal absorption in mammals Is minimal, possibly as
little
as 5% of total dietary beta-sitosterol consumed. In contrast, intestinal
absorption of
cholesterol is 45-54% of intake. Unlike cholesterol, beta-sitosterol is
secreted into the
bile and is esterified outside the intestinal wall at a much slower rate.
After secretion
into the bile, beta-sitosterol is stored in the gallbladder, then released
intermittently
into the duodenum, and subsequently incorporated into feces.
It has been proposed that phytosterols inhibit the uptake of dietary and
endogenously produced cholesterol from the gut, causing a decrease in serum
cholesterol levels (Nguyen 1999). One theory suggests that cholesterol in the
intestine, already marginally soluble, is precipitated into a non-absorbable
state by the
presence of added phytosterols.(Hicks and Moreau 2001). A second theory
proposes
that cholesterol must enter bile salt and phospholipid containing "mixed
micelles" to
be absorbed into the bloodstream. Cholesterol is only marginally soluble in
these
micelles and is displaced by phytosterols, preventing its absorption (Hicks
and
Moreau 2001). Due to the limited capacity in the micelles for carrying
cholesterol,
compounds with similar structures, such as plant sterols, can compete with
cholesterol for this space. Therefore, increasing the amounts of plant sterols
may
result in less cholesterol in mixed micelles and hence, decreased absorption
of
cholesterol from the gut.
Orally ingested flavonolds are largely present as aglycons In the intestine
and
become absorbed with micelles of bile acids into the epithelium and then into
the
blood. Through the portal vein, the major part of the flavonoids would be
delivered to
CA 02538494 2006-02-24
the liver, which decomposes them (Havsteen 1983, Biachem Pharmacol 32: 1141-
1148).
Both EPA and DHA have been found to alter plasma membrane composition,
cell-signaling mechanisms, eicosenoid responses, eytokine release, and immune
cell
5 responses (Arsian at al, 2002, Lipids 37: 935-941). One mechanism for these
effects
may be that EPA and DHA are incorporated into membrane phospholipids by
replacing arachidonic acid (AA, n-6 fatty acid). AA is the substrate for the
synthesis of
eicosanoids, such as fluomboxane A2 and prostaglandin E2 (O'Morain at al.
1990,
Scand J Gastroenterol 31: 267-272). N-3 fatty acids have a greater affinity
for the
10 cyclo- and lipoxygenase enzymes than n-6 fatty acids and they competitively
inhibit
the formation of prostaglandins and leukotrienes (Drevon 1992). Therefore,
increased
dietary intake of n-3 fatty acids may shift the balance of the eicosanoid
production to a
less inflammatory profile (Arsian et al, 2002). Another mechanism for the
effect of n-3
fatty acids is associated with the change in fluidity by incorporation of
fatty acids in the
15 cell membranes and an influence on the activities of membrane-associated
enzymes
or receptors (Vognild et al. 1998, Lipids 33: 427-436).
This study was designed to investigate whether supplementation with the
above-described supplement comprising plant sterols, antioxidants and
essential fatty
acids could, in subjects with non-food allergies.
a) Decrease Immunological response to allergens
b) Beneficially modify blood lipid parameters
Subjects
Twenty subjects consisting of 16 females and 4 males, aged from 20-50, and who
fit
specific inclusion/exclusion criteria were recruited, The inclusion/exclusion
criteria
were as follows:
Inclusion Criteria:
= Male and female subjects aged 18 years or older
CA 02538494 2006-02-24
16
= For females, confirmation (via pregnancy test) of non-pregnancy at baseline,
and
use of acceptable birth control method in females of childbearing potential
= No food allergies
= Willing and able to give informed written consent
Exclusion Criteria:
= Pregnancy/breast feeding
= History of clinical significant and unstable cardiovascular, pulmonary,
renal,
neurological dermatological, hepatic or endocrine disease in the past 6 months
= Change in medication 4 weeks before entry into study
= History of frequent respiratory infections
= Diabetes or immune disorder such as lupus erythematosis or HIV/ AIDS
= Known allergy to the supplement or any of its components
= History of drug, alcohol or substance abuse in the past 5 months
= Participation in any clinical trial within 6 weeks preceding day 1 of the
study
All participants were recruited from the University of Guelph or the City of
Guelph. At the study screening, participants were given a letter of
information and the
consent form to sign. Respondents received a verbal briefing of the study
protocol
and the same information in writing. They signed an informed consent and
completed
a questionnaire to ensure compliance with the inclusion/exclusion criteria.
All participants were questioned on the nature of their allergies. Eligible
subjects were required to have non-food allergies, but were required to have
allergies
to dust, animal dander, animal saliva, molds, etc. In addition, the study
required that
all subjects had received an allergy test by their physician to verify
allergen response,
or had been prescribed medication in the past to treat their allergies.
Experimental Design
This study was conducted as a double blind, placebo-controlled clinical trial.
Subjects in the treatment group consumed two capsules of the above-described
supplement for the first seven days In order.to reach phytosterol threshold
levels,
CA 02538494 2006-02-24
17
followed by a maintenance dosage of 1 capsule per day. In these experiments,
each
capsule contained 300mg of phytosterols (117mg of j -Sitosterol) derived from
soy,
20mg of EnzogenolN (antioxidants) extracted from pine bark, and 50mg of
Cellasatel' (a mixture of proteins and essential fatty acids) from seeds and
fish oils,
although as will be appreciated by one of skill in the art, other suitable
formulations as
discussed above may also be used. These include embodiments wherein the
composition or supplement comprises: 10-90% phytosterols, 5-85% essential
fatty
acid complexes, and 5 to 85% antioxidant; or 29% to 75% phytosterols; 10% to
50%
essential fatty acid complexes; and 5% to 50% antioxidants as well as
embodiments
wherein the supplement or composition comprises 29% to 75% beta-sitosterol;
10%
to 50% essential fatty acid complexes; and 5% to 50% antioxidants as well as
other
embodiments described herein.
The placebo consisted of a rice flour mixture, identical in appearance to the
supplement, and the consumption pattern for the placebo was the same for the
placebo group as for the treatment group. The supplement and placebo
supplementation phase lasted 28 days.
Subjects were required to visit the HNRU human clinical testing unit on three
separate occasions. Subjects came in for baseline testing (day 0) and returned
on day
7 of the study and again on day 28, the last day of the study. On the initial
visit,
height, weight and a blood sample were taken. Blood was analyzed for complete
blood cell count (CHC), plasma DHEA, Cortisol, total HDL and LDL-cholesterol
concentrations, and triglycerides. A Quantikine high sensitivity Elisa kit was
used in
order to measure human IL-6 in the plasma collected. Throughout the study,
subjects
were required to complete a daily journal listing specific allergy symptoms,
non-allergy
symptoms and any medications taken during the supplementation phase.
RESULTS
Immune Parameters
Basophils
CA 02538494 2006-02-24
18
The effects of the supplement on immune parameters are presented in Table
1. A number of studies support the belief that human basophils play an
important role
in allergic inflammation. Mast cells and basophils express the high affinity
receptor for
IgE (FcepsilonRl) and play a central role for IgE-associated immediate
hypersensitivity reactions and allergic disorders. During allergic reactions,
basophils
migrate from the blood compartment to inflammatory sites, where they act as
effector
cells in concert with eosinophils. Basophils release histamine during
inflammation and
allergic reactions.
The participants in the treatment group, when compared to the control group,
showed a significant reduction in basophil count, while the reduction seen in
the
control group was non-significant. A reduction in basophil count may indicate
a
reduction in histamine release.
lntetleukin-6
The immune system also responds to stressors by causing certain immune
cells to secrete the pro-inflammatory cytokines, Interleukin-1 (IL-1) and
Interleukin-6
(IL-6). These cytokines are both involved in inflammation and IL-8 in
particular is
thought to worsen the symptoms of autoimmune diseases and fibromyalgia.
Interleukin-6 has been found to act as a growth factor in several tumors and
some
viruses also use IL-6 to replicate. Interleukin-6 also causes calcium to be
released
from bone, promoting osteoporosis. We must control the release of these
cytokines if
we want to enhance immunity and reduce degenerative diseases.
It was noted in the pilot trial that the pro-inflammatory cytokine IL-6 levels
showed a substantial reduction in the treated group when compared to the
control
group,
The supplement has demonstrated that it has an effect on histamine-containing
basophil counts and a reduction of IL-6 levels, and consequently may
substantially
alleviate symptoms associated with airborne allergens, asthma and allergic
rhinitis.
CortisollDHEA Levels
CA 02538494 2006-02-24
19
The body has developed mechanisms to protect it from the damaging effects of
stress. The "fight-or-flight" response is one way the body deals with extreme
situations of stress. Upon realizing we are in danger, the brain sounds an
alarm,
telling our adrenal glands to secrete adrenaline and cortisol, which mobilizes
the body
to fight or run. This response is supposed to be a short-lived reaction yet
today most
of us are In and out of this state continually. As a result, our immune system
becomes
imbalanced, sending out too many inflammatory cytokines. Our adrenal glands
become exhausted, weakening several body systems, especially the
cardiovascular
and endocrine systems.
DHEA is an abbreviation for dehydroeplandrosterone. It is a hormone made
primarily by the adrenal or stress glands. Hormones are messenger molecules
that
influence the function of cells and tissues all over the body. DHEA and
cortisol are the
body's long-acting stress hormones and are antagonistic to each other to some
degree. Whereas DHEA has an anabolic or building influence, cortisol has a
catabolic
or tearing down effect on the body. Both of these effects are essential and
these two
hormones must be in proper balance for optimal health. How do these hormones
become imbalanced?
By stress maladaptation. Stress maladaptation is the body's inappropriate
response to prolonged stress. The normal reaction of the body to stress is to
produce
greater quantities of both cortisol and RHEA. When the stress is gone, the
body
reduces its output of cortisol and DHEA to resting levels and everything is
fine. This is
what happens with short episodes of stress. However, when the stress is
prolonged,
the body prefers to make increasingly greater amounts of cortisol and less
DHEA.
How long does it take for this to occur?
One study showed that after just 28 days of continuous stress cortisol levels
had climbed to 240 percent of starting values and DHEA had dropped to 15
percent of
initial levels, What's even worse is that even after the stress is removed,
the body
sometimes does not recover and bring these hormones back to normal levels, but
instead, remains in the stress response mode with high cortisol and low DHEA
output.
The consequences of elevated cortisol and reduced DHEA levels are devastating:
the
CA 02538494 2006-02-24
immune system is compromised with increased risk to infections, certain
cancers,
allergies and autoimmune diseases.
A tremendous body of research has shown that when cortisol goes up, DHEA
drops and when DHEA is normal, cortisol also normalizes. Low DHEA levels are
seen
5 in those that are immune compromised, have arteriosclerosis (hardening of
the
arteries), diabetes and lupus.
Cortisol helps the body maintain homeostasis in the face of stressors,
counteracts inflammatory and allergic reactions and controls the metabolism of
protein and carbohydrates. Cortisol Is a very misunderstood hormone. Balance
is the
10 key. In naturally low doses, it stimulates the immune system and in high
doses, as
prescribed in synthetic drug form, it can be immune suppressing. Remember that
cortisol plays a role in counteracting inflammatory responses in the immune
system
and when cortisol is not available because the adrenal glands have become
exhausted from too much stress, inflammation is allowed to continue unchecked.
15 Conversely, too much Cortisol and you have immune suppression.
In the conventional standard of care, any cortisol level within a very broad
range is considered normal, and anything outside that range indicates disease.
Cortisol production has an ACTH dependant circadian rhythm with peak levels in
the
early morning and a nadir at night (salivary cortisol ranges can vary from 8.0
to 1.0 in
20 the morning and 1.0 to 0.1 in the evening) The factor controlling this
rhythm is not
completely defined and can be disrupted by a number of physical and
psychological
conditions. ACTH and cortisol are secreted independent of circadian rhythm in
response to physical and psychological stress.
In the early stages of adrenal stress, cortisol levels will be too high during
the
day and continue rising in the evening. This is called "hyperadrenia". In the
middle
stages, cortisol may rise and fall unevenly as the body struggles to balance
itself
despite the disruptions of caffeine, carbs and other factors, but levels are
not normal
and are typically too high at night. In advanced stages, when the adrenals are
exhausted from overwork, cortisol will never reach normal levels
("hypoadrenia").
CA 02538494 2006-02-24
21
None of the participants in the trial were known to have any of the autoimmune
diseases that are associated with elevated cortisol levels. The change in
cortisol
levels noted in the pilot trial appear to be in the normal range, and neither
DHEA nor
cortisol levels, nor the ratio of these two parameters, showed significant
changes at
the pa0.05 level.
Carrliovascutar parameters
The effects of the supplement on lipid and lipoprotein parameters are
illustrated
in Table 2 and Table 3.
Cholesterol is an extremely important biological molecule that has roles in
membrane structure as well as being a precursor for the synthesis of the
steroid
hormones and bile acids. Both dietary cholesterol and that synthesized de novo
are
transported through the circulation in lipoprotein particles. The same is true
of
cholesterol esters, the form In which cholesterol is stored in cells.
The synthesis and utilization of cholesterol must be tightly regulated In
order to
prevent over-accumulation and abnormal deposition within the body. Of
particular
importance clinically is the abnormal deposition of cholesterol and
cholesterol-rich
lipoproteins in the coronary arteries. Such deposition, eventually leading to
atherosclerosis, is the leading contributory factor in coronary artery
disease.
Cholesterol is minimally soluble in water; it cannot dissolve and travel in
the
water-based blood stream. Instead, it is transported in the blood stream by
lipoproteins, i.e. protein "molecular-suitcases" which are water soluble and
carry
cholesterol and fats internally. The proteins form part of the surface of the
given
lipoprotein particle and determine from what cells cholesterol will be removed
and
where it will be supplied.
The largest lipoproteins, which primarily transport fats from the intestinal
mucose to the liver, are called chylomicrons. They carry mostly triglyceride
fats and
cholesterol (both from food and especially internal cholesterol secreted by
the liver
into the bile). In the liver, chylomicron particles give up triglycerides and
some
cholesterol and are converted into low-density lipoprotein (LDL) particles
which carry
CA 02538494 2006-02-24
22
triglycerides and cholesterol on to other body cells. In healthy individuals
the
low-density lipoprotein (LDL) particles are large and relatively few in
number.
Conversely, high numbers of small low-density lipoprotein (LDL) particles are
strongly
associated with promoting atheromatous disease within the arteries.
High-density lipoprotein. (HDL) particles transport cholesterol back to the
liver
for excretion, but vary considerably in their effectiveness for doing this.
Having large
numbers of large HDL particles correlates with better health outcomes.
Conversely,
having small amounts of large HDL particles is strongly associated with
atheromatous
disease progression within the arteries.
The cholesterol in LDL cholesterol and the cholesterol in HDL cholesterol are
identical. The only difference between the two is the carrier protein
molecules (i.e. the
lipoprotein).
High cholesterol levels has been shown in many trials to be the source of
cardiovacular disease. High cholesterol is the best known of all the many
threats to a
healthy heart. When excess amounts of this waxy, fat-like substance build up
along
the walls of the arteries, you face a dramatically higher risk of a complete
blockage,
leading to a heart attack or stroke.
At normal levels, cholesterol Is not a bad thing. On the contrary, it's an
essential raw material used by the body to build cell walls and produce
hormones
such as estrogen and testosterone. The body produces its own supply of
cholesterol
in the liver, and its found naturally in all animal products (such as meats,
eggs, milk,
and cheese). It poses a problem only when the body is unable to use or
eliminate
excessive supplies.
As one of a variety of fatty substances in the body, cholesterol is classified
as a
lipid. It is carried through the bloodstream attached to proteins, forming
complexes
called lipoproteins. There are two major types of lipoproteins: the low-
density
lipoproteins (LDL) commonly known as "bad" cholesterol, and the high-density
lipoproteins (HDL) usually dubbed "good" cholesterol. Its the "bad" LDL
cholesterol
that tends to form deposits on the artery walls. HDLs, on the other hand, help
to clear
CA 02538494 2006-02-24
23
excess cholesterol from the bloodstream. The ideal situation to aim for then,
is a low
level of LDL cholesterol, a high level of HDL cholesterol, and a moderate
total of both.
The specific objective of this portion of the trial was to determine the
effects of
the supplement on blood lipid parameters. Significant reduction was noted in
the
overall LDL levels of the treatment group from day 0 to day 28. Perhaps what
is more
interesting is the increase in HDL levels, compared with a relative decrease
in the
placebo group.
However it is the ratios of various lipids and lipid proteins rather than the
absolute values that are important in assessing cardiovascular risk, and
consequently
these ratios were calculated and tabulated.
A significant decrease in the ratio of TC/HDL, and in the ratio of LDL/HDL
cholesterol in the supplement group was noted. A decrease in these ratios
corresponds to an associated decrease in the risk of cardiovascular disease
(CVD).
These ratios are markers for a reduction in the risk of developing
atherosclerosis.
Consequently these results indicate that the supplement is very beneficial to
for
example the health of hypercholesterolemic individuals at risk of developing
CVD.
As discussed above, there is provided a method of modifying blood lipid
parameters comprising administering to an individual in need of such treatment
an
effective amount of the composition or supplement as described herein. As will
be
apparent to one of skill in the art, an effective amount of the supplement
refers to the
quantity thereof necessary to reduce the TC/HDL ratio or reduce the LDLJHDL
ratio of
an individual in need or desirous of such treatment and such dosages and
regimens
are enabled herein.
Specifically, as can be seen in table 2, the blood lipid parameters of those
in
the supplement group were modified in that total cholesterol was reduced by
5.3%,
low density lipoprotein was reduced by 15% and HDL was increased by 4.3%. This
is
in stark contrast with the results observed for the control group fed the
placebo, also
shown in Table 2, wherein total cholesterol actually increased by 8.2%, LDL
increased
by 0.7% and HDL decreased by 4.7%.
CA 02538494 2012-05-23
24
As summarized on Table 3, the test group administered the supplement or
composition showed a TC/HDL ratio reduced by 9.4% whereas the control or
placebo
group had an increased TC/HDL ratio of 1.7%. Similarly, the supplement group
showed an LDUHDL ratio decrease of 17.7% whereas the placebo or control group
had only a 0.5% decrease. Finally, the supplement group showed a total
glycerides to
HDL increase of 3.0% whereas the control group showed a 20% increase.
Thus, administration of the supplement as described herein has been shown to
reduce total cholesterol, increase HDL, decrease LDL, reduce the TC/HDL ratio
and
decrease the LDUHDL ratio in test individuals compared to individuals
administered a
placebo or control. As such, the supplement can be used to reduce total
cholesterol,
increase HDL, decrease LDL, reduce the TC/HDL ratio and decrease the LDUHDL
ratio.
The supplement has an effect on immune parameters and, in particular, on
basophil and possibly IL-6 levels. Given these changes, the supplement has the
potential to substantially alleviate allergic responses.
The supplement could also have an effect in autoimmune diseases such as
Crohn's disease or rheumatoid arthritis, or in the ability of subjects to
resist the
common cold virus.
This study verified that the supplement is effective in reducing circulating
levels
of LDL-cholesterol and increasing circulating levels of HDL cholesterol. It is
of interest
to note that there was a significant decrease in the ratio of TC/HDL, and in
the ratio of
LDUHDL cholesterol, in the supplement group. A decrease in these ratios
corresponds to an associated decrease in cardiovascular disease (CVD) risk,
because these ratios are markers for a reduction in the risk of developing
atherosclerosis.
CA 02538494 2006-02-24
REFERENCES
International Journal of Immuno Pharmacology 1996 Vol 18 Bouic at al.
5 Beta-sitosterol and beta-sitosterof glucoside, stimulate human peripheral
blood
lymphocyte proliferation; implications for their use as an immunomodulatory
vitamin
combination.
International Journal Molecular Medicine 2000 Vol 5 Awad at al. Inhibition of
growth,
10 and stimulation of apoptosis, by beta-sitosterol treatment of human breast
cancer
cells.
Anti Cancer Research 1998 Vol 18 (1 a) Awad, Holtz et al. Beta-sitosterof
inhibits
growth of HT-29 human colon cancer cells.
Nutrition and Cancer 1998 Vol 32 (1) von Holtz, Fink et al. Beta-sitosterol
induces
apoptosis in LNCaP human prostate cancer cells.
AIDS Bulletin 1997 Vol 6 Bouic. Immodulation in HIV/AIDS: The
Tygerberg/Stellenbosch University Experience.
South African Journal of Science, 1997 Vol 93 Pegel. The Importance of
Sitosterol
and Sitosterolin in human and animal nutrition.
British Journal of Urology 1997 Vol 80 Klippel et al. A multi centric, placebo
controlled,
double blind clinical trial of betasitosterol for the treatment of benign
prostatic
hyperplasia,
The Lancet 1995 Vol 345 Berges at al. Randomized placebo controlled double
blind
clinical trial of beta-sitosterol in patients with benign prostatic
hyperplasia.
CA 02538494 2006-02-24
26
The Arthritis Trust of America Summer 98 Bouic. Sterols/Sterolins the natural,
none
toxic immuno-modulators and their role in the control of rheumatoid arthritis.
Nutrition Reviews 1992 Vol 50 NO. 7 Block. The data support a role for
antioxidants in
reducing cancer risks.
Methods in Enzymology 1990 Vol 186 Bors, Heller et al. Flavanoids as anti-
oxidants:
determination of radical scavenging efficiencies.
The Lancet 1998 Vol 344 Cerruti. Oxi-radicals and cancer.
Biochemical Pharmacology 1990 Vol 39 No. 11 DeWhalley, Rankin et al.
Flavanoids
inhibit the oxidative modification of low density lipoproteins by macrophages.
The Lancet 1994 Vol 344 Grishan. Oxidants and free radicals in inflammatory
bowel
disease.
Phytochemistry 1987 Vol 26 No. 9 Husain, Cillard et al. Hydroxyl radical
scavenging
activity of flavanoids.
The Lancet 1994 Vol 344, Jenner. Oxidative damage in neuerodagenerative
disease.
Free Radicals in Diagnostic Medicine, D. Armstrong, Plenum Press New York.
Free radical scavenging and antioxidant activity of plant flavanoids.
New Zealand Journal of Science 1973 Vol 16 Markam and Porter. Extractives of
pinus
radiate bark- phenolic components.
New Zealand Journal of Science 1974 Vol.17 Porter, Extractives of pinus
radiate
bark-procyanidin constituents.
CA 02538494 2006-02-24
27
Free radical Research 1995 Vol 22 No. 4 Rice-Evens, Miller et al. The relative
antioxidant activities of plant derived polyphenolic flavanoids.
Biochemical Pharmacology 1998 Vol 37 No 5 Rbak, Gryglewski. Flavanoids are
scavengers of super oxide anions.
Free Radical Biology in Medicine 1990 Vol 9 Muting, Rongliang et al.
Flavanoids as
super oxide scavengers and antioxidants.
CA 02538494 2006-02-24
28
Table 1. The effects of supplement on specific immune parameters in
experimental
and placebo groups from day 0 to day 28
Immune Supplement Supplement Control Control
Parameter Day 0 Day 28 Day 0 Day 28
IgE 472.00 451.00 1335.00 1127.00
DHEA 6.44 6.44 4.93 4.77
Cortisol 507.00 584.00 490.00 498.00
Cortisol/DHEA 94.06 108.36 160.66 141.44
IL-6 1.261 0.937 1.318 1,179
WBC 7.41 7,24 7.28 7,13
Lymphocyte 2.16 2.24 2.56 2.60
Segmented
Neutrophil 4.65 4.39 4.11 3.97
Monocytes 0.33 0.34 0.35 0.31
Eosinophils 0.24 0.20 0.23 0.20
Basophils 0.23 0.01 0.13 0.04
Table 2 The effects of supplement on blood lipid parameters in experimental
and
placebo groups from day 0 to day 28
Blood Lipid Supplement Supplement Control Control
Parameter Day 0 Day 28 Day 0 Day 28
Total Cholesterol 4.36 4.13 4.87 4.91
LOL 2.27 1.93 2.85 2.87
HDL 1.63 1.70 1.48 1.41
TG 1.00 1.09 1.18 1.38
CA 02538494 2006-02-24
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Table 3. The effects of supplement on specific cardiovascular ratios in
experimental
and placebo groups from day 0 to day 28
Cardiovascular
Parameter Supplement Supplement Control Control
Ratios Day 0 Day 28 Day 0 Day 28
TC/HDL 2.88 2.61 3.50 3.56
LDLIHDL 1.58 1.30 2.11 2.10
TG/HDL 0.66 0.68 0.85 1.02
