U.S. patent application number 14/698659 was filed with the patent office on 2016-08-25 for bioactive-rich concentrates and nutritive and therapeutic products containing same.
The applicant listed for this patent is Kartik Natarajan. Invention is credited to Aravind Cherukuri, Reddy Sastry Cherukuri, Rukmini Cheruvanky, Kartik Natarajan.
Application Number | 20160243185 14/698659 |
Document ID | / |
Family ID | 38605091 |
Filed Date | 2016-08-25 |
United States Patent
Application |
20160243185 |
Kind Code |
A1 |
Natarajan; Kartik ; et
al. |
August 25, 2016 |
Bioactive-rich concentrates and nutritive and therapeutic products
containing same
Abstract
This invention relates generally to the field of compositions
for use as nutraceuticals, food additives or adjuncts to
conventional drug therapies. In particular, the invention relates
to compositions derived from natural oil sources which can be used
for effective and inexpensive treatment of cardiovascular diseases,
hypercholesterolemia, diabetes, cerebrovascular disease,
neurological disorders, or liver abnormalities.
Inventors: |
Natarajan; Kartik; (West
Hartford, CT) ; Cherukuri; Aravind; (Newton, MA)
; Cheruvanky; Rukmini; (Folsom, CA) ; Cherukuri;
Reddy Sastry; (Folsom, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Natarajan; Kartik |
West Hartford |
CT |
US |
|
|
Family ID: |
38605091 |
Appl. No.: |
14/698659 |
Filed: |
April 28, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11406068 |
Apr 18, 2006 |
9034919 |
|
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14698659 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23V 2002/00 20130101;
A23L 33/115 20160801; A61K 31/401 20130101; A61K 31/56 20130101;
A61K 36/00 20130101; A61K 45/06 20130101; A23L 33/11 20160801; C07D
311/72 20130101; A23L 7/117 20160801; A61K 31/22 20130101; A61K
35/00 20130101; A23L 7/126 20160801; A61K 36/899 20130101; A61K
31/366 20130101 |
International
Class: |
A61K 36/899 20060101
A61K036/899; A61K 45/06 20060101 A61K045/06 |
Claims
1. A bioactive-rich concentrate derived from the unsaponifiable
fraction of an oil of rice bran or rice germ, consisting
essentially of the unsaponifiable content of said oil,
substantially free of rice bran contaminants; the unsaponifiable
content being present in said concentrate at a concentration 10 to
100 times greater than in rice bran oil or rice germ oil.
2. The concentrate of claim 1, wherein the unsaponifiable contents
comprises a 4-dimethyl sterol component, a 4-monomethyl sterol
component, a .gamma.-oryzanol component, a polyphenol component, a
tocopherol component, and a tocotrienol component.
3. The concentrate of claim 2, wherein the amount of 4-dimethyl
sterol component is 10-30%, the amount of 4-monomethyl sterol
component is 4-14%, the amount of .gamma.-oryzanol component is
20-40%, the amount of polyphenol component is 5-15%, the amount of
tocopherol component is 5-20%, and the amount of tocotrienol
component is 5-20%; by weight; based on total weight of the
concentrate.
4. A composition comprising a bioactive-rich bioactive concentrate
of claim 1 in combination with at least one of: a cholesterol
lowering drug, herbs, and/or an HMG CoA reductase inhibitor.
5-7. (canceled)
8. The composition of claim 4, wherein the HMG CoA reductase
inhibitor is selected from the group consisting of mevastatin,
lovastatin, pravastatin, simvastatin, fluvastatin, cerivastatin,
atorvastatin, tenivastatin, rosuvastatin, pitavastatin and
combinations thereof.
9. The composition of claim 4, further comprising one or more
pharmaceutically acceptable excipients.
10. (canceled)
11. The composition of claim 4, wherein the composition is intended
for oral administration.
12. The composition of claim 11, wherein the composition is in a
form selected from the group consisting of tablets, capsules,
liquids, suspensions, solutions, and emulsions.
13. The composition of claim 4, wherein the cholesterol lowering
drug is a bile acid sequestrant or a fibric acid derivative.
14. The composition of claim 13, wherein the bile acid sequestrant
is selected from the group consisting of cholestyramine,
colesevelam and colestipol.
15. The composition of claim 13, wherein the fibric acid derivative
is selected from the group consisting of fenofibrate and
gemfibrozil.
16. A method of treating a patient in need of lowering serum
lipids, cholesterol, blood glucose and/or triglycerides, comprising
administering to the patient the bioactive-rich concentrate of
claim 1 in a dosage regime effective to lower the patient's serum
lipid level.
17. A method of treating a patient in need of lowering serum
lipids, cholesterol, blood glucose and/or triglycerides, comprising
administering to the patient the combination product of claim 4 in
dosage regime effective to lower the patient's serum lipid
level.
18. A method of treating a patient in need of increasing HDL-C
levels, comprising administering to the patient the combination
product of claim 4 in a dosage regime sufficient to increase the
patient's HDL-C level.
19. The method of claim 17, wherein the components of the
combination therapy are simultaneously administered.
20-21. (canceled)
22. A method of treating a patient in need of treatment for
hypertension, hyperlipidemia, obesity, inflammatory disease,
arthritis, hypercholesterolemia, cardiovascular disease,
cerebrovascular disease, arteriosclerosis, diabetes mellitus,
immune dysfunction or cancer, neurological disorders, or liver
abnormalities comprising administering to the patient the
bioactive-rich concentrate of claim 1, in a dosage regime effective
to treat the patient.
23. A method of treating a patient in need of treatment for total
serum cholesterol, LDL-C, apolipoprotein B, triglycerides,
improving HDL-C levels, inhibiting platelet aggregation and
dissolving aortic streaks, comprising administering to the patient
the bioactive-rich concentrate of claim 1, in a dosage regime
effective to treat the patient.
24. A food product comprising the bioactive-rich concentrate of
claim 1 and a food substance.
25-27. (canceled)
28. The food product of claim 22, which contains from about 0.1% to
15% of the bioactiverich concentrate.
29. The bioactive-rich concentrate of claim 1, wherein said
bioactive-rich concentrate is in its natural matrix.
30-31. (canceled)
Description
FIELD OF THE INVENTION
[0001] This invention relates generally to the field of
compositions for use as nutraceuticals, food additives or adjuncts
to conventional drug therapies. In particular, the invention
relates to compositions derived from natural oil sources which can
be used for effective and inexpensive prophylaxis of cardiovascular
diseases, hypercholesterolemia and/or diabetes, and management of
lipid metabolism.
BACKGROUND
[0002] Natural phytonutrients, bioactives and antioxidants
("bioactives") are used in the nutraceutical and pharmaceutical
industries for their health benefits. For example, polyphenols from
grape seed, pine bark and garlic are used in nutraceutical
formulations. There are also several potent bioactives present in
rice bran and rice germ oil. The unsaponifiable fraction (i.e., the
non-fat portion) of rice oil has a high concentration of useful
micronutrients and antioxidants such as tocopherols, tocotrienols,
.gamma.-oryzanol, phytosterols, polyphenols and squalene, when
compared to other vegetable oils. Several clinical studies with
rice bran oil (RBO) demonstrated significant hypocholesterolemic,
hypolipidemic and anti-atherogenic properties; see, e.g., Table
1.
TABLE-US-00001 TABLE 1 Selected RBO clinical studies Methodology
Parameters Reading (mg/dl) Change (%) Reference Rice Bran Oil at 60
g/day Total 219-177 -19 Lichtenstein, A. H, et al. 15 moderately
Cholesterol (1994) "Rice Bran oil hypercholesterolomic LDL-C
162-122 -25 consumption and plasma subjects. HDL-C no change no
change lipid levels in moderately 32 days, double blind
Triglycerides 131-109 -17 hyper cholesterolemic crossover latin
square design humans." Arterioscelorsis along with canola, corn and
& Thromobosis v. 14 olive oils. 549556 Rice Bran Oil at 35
g/day. Total 247-183 -25 Raghuram, T. et al. (1989) 12
hyper-cholesterolemic Cholesterol "Studies on hypolipidemic
subjects. Triglycerides 349-212 -35 effects of dietary Rice 30
days, with a control group Bran Oils in humans." of 9
hyper-cholesterolemic Nutrition Reports subjects with peanut oil.
International v. 39(5): 889-895 Rice Bran Oil at 60 g/day. Total
194-164 -15 Suzuki, S. & Oshima, S. 50 healthy females of
normal Cholesterol (1970) "Influence of cholesterol. Total 164-121
-26 blending of edible fat and 7 days without blend. Cholesterol
oils on serum cholesterol 7 days blend with Safflower Total 194-164
-19 levels. Japanese J. Oil (SO). Cholesterol Nutrition v. 28 (1):
Part 1 At RBO:SO of 70:30 pp. 3-6; Part 2 pp. 194-196. At RBO:SO of
85:15
[0003] These beneficial properties are attributed to the potent
phytonutrients, micronutrients and antioxidants present in the
unsaponifiable fraction of the oil (see, e.g., Table 2.) The
individual constituents of the rice bran oil unsaponifiable
fraction have been well studied for their hypolipidemic,
hypoglycemic, hypocholesterolemic, antioxidant and other health
benefits in animals and in human subjects.
TABLE-US-00002 TABLE 2 Selected study on RBO unsaponifiable
fraction Readings Final Initial USF/RBO Methodology Lipid
Parameters (mg/dl) (mg/dl) Change % References 0.4% RBO
Un-Saponifiable Total Cholesterol 374 243/288 -35/-23 Sharma, S. D.
and Fraction (USF) vs. LDL/VLDL 331 195/240 -41/-27.5 Rukmini, C.;
Indian J. equivalent 10% Rice Bran HDL 43 48/48 11.6/11.6 Med. Res.
1987 Mar; Oil (RBO) studied in rates 85: 276-81 for
hypcholesterolemic effect Unsaponifiable fraction: Total
cholesterol, No change Ha. T., Han, S., Kim, S., 277.7 mg/g
.gamma.-oryzanol, 89.99 mg/g Triglycerides No change Kim, I., Lee,
H., and phytosterols, 11.66 mg/g HDL-C Increase Kim, H., (2005)
tocols, 3.36 mg/g Nutrition Research 25: squalene, 0.73 mg/g of
597-606. octocosanol Unsaponifiable fraction Total cholesterol
Decreased Lee, J., Lee, S., Kim,. M., LDL-C Decreased Rhee, C.,
Kim, I., Triglycerides Decreased and Lee, K. (2005) J. HDL-C No
change Sci. Food. Agri 85: 493-498.
[0004] Antioxidant defense mechanisms in biological systems play a
major role in the prevention of a number of diseases such as
cardiovascular, cerebrovascular, carcinogenic, and other metabolic
age-related disorders. Free radicals, such as singlet oxygen, are
highly reactive, attack cellular components, can damage DNA, and
alter normal metabolism, resulting in a disease state. Humans are
under constant challenge by free radicals; unless charged with
sufficient antioxidants to quench these free radicals, the pace of
damage to the body is increased and can result in various disease
states. Antioxidants provide a defense mechanism and help in
preventing and arresting the progression of diseases. It is a
constant battle to maintain the delicate balance between oxidants
and antioxidants in the body. Epidemiological evidence is mounting
on the significant role of natural antioxidants and their vital
role in maintaining health and preventing diseases.
[0005] Cardiovascular disease (CVD) is a leading cause of mortality
in the United States and in developed countries around the world.
The disease is linked with well-defined risk factors, such as lipid
anomalies, arterial hypertension, diabetes, obesity and smoking.
The estimated breakdown of the CVD patient population is as
follows: High blood pressure: 50.0 million; Coronary heart disease:
13.9 million; Congestive heart failure: 4.7 million; Stroke: 4.0
million; Rheumatic heart disease: 1.8 million.
[0006] There is an increasing need to contain this disease
effectively without exponentially increasing the associated
healthcare costs. Prescription medications alone will not suffice
in addressing this need as they have a high direct and indirect
cost. Beyond cost, there are also significant side effects
associated with prescription drugs and medical practitioners are
reluctant to increase dosages unless absolutely needed. For
example, statin drugs for lowering cholesterol are effective, but
can have undesired side effects.
[0007] Rice bran oil (RBO) is obtained from rice bran, i.e., the
mesocarp of paddy. RBO is different from other vegetable oils,
which are obtained from the seed and/or nuts. Palm oil, coconut
oil, olive oil and rice bran oil are obtained from the mesocarp of
the fruit. These oils are rich in several natural antioxidants.
However, when these oils are processed to an edible grade during
normal refining steps, the valuable bioactive micronutrients and
antioxidants contained in the unsaponifiable fraction of the oil
are degraded or destroyed during normal refining steps, thus
reducing both the bioactivity of the micronutrients and
antioxidants and their therapeutic benefit. Obtaining a bioactive
micronutrient and/or antioxidant-rich rice oil derivate has posed
significant challenges to the oil technologist.
[0008] Conventional edible oil refining processes yield several
by-products, including soap stock, distillate and gums. These
by-products contain high concentrations of several of the vegetable
oils' unsaponifiable constituents. Although technologies are
available for isolating desirable individual constituents and
actives, e.g., tocopherols, tocotrienols and .gamma.-oryzanol,
etc., in the unsaponifiable fractions of rice bran and other
vegetable oils, the art has not advanced far enough in developing
therapeutic applications of suitable natural whole food extract
concentrates containing such micronutrients, e.g., in the
unsaponifiable fraction of vegetable oils.
[0009] Although the components of the unsaponifiable fraction of
rice bran oil has been reported for its cholesterol reducing
property, the dose at which the individual components give the
desired effect is very high. Rice bran oil and palm oil is the
predominant natural source of tocopherols and tocotrienols
(tocols), however, phytosterols are available in several vegetable
oils.
[0010] .gamma.-oryzanol is unique to rice bran oil among natural
products. It occurs in the highest concentration in the bioactives
of the unsaponifiable fraction of rice bran oil. .gamma.-oryzanol
is a mixture of at least five components. These five components
include ferulic acid esters of cycloartenol, 24-methylene
cycloartanol, .beta.-sitosterol, 4-methylsterol and methyl
ferulate. .gamma.-oryzanol is a potent antioxidant (Joseph Bruno
"The facts on .gamma.-oryzanol" 1987), demonstrated to have
UV-quenching properties, and activity on the hypothalamus,
relieving stress by increasing DHEA levels and reducing cortisol
levels. In addition, it has been reported to induce fat burning
that results in lean body mass. Furthermore, it has been shown to
have hypocholesterolemic activity, dissolving aortic streaks
(Seetharamiah, G. et al., Atherosclerosis 78: 219-223) and
dissolving blood clots. It has also demonstrated beneficial effects
on cardiovascular risk factors.
[0011] U.S. Pat. No. 5,660,691 discusses isolating tocopherols and
tocotrienols from rice distillate. U.S. Pat. No. 5,288,902
discusses isolating .gamma.-oryzanol, another valuable antioxidant,
from the soap stock, which is typically discarded as a waste
product. However, in these techniques and others, the natural
matrix is disrupted, and thus the potency and bioavailability of
these components is diminished.
SUMMARY
[0012] The present invention provides effective and non-destructive
isolation, concentration and use of the bioactive components, in
their natural matrix, from the unsaponifiable fraction of rice bran
oil or rice germ oil. The importance of the effective isolation and
concentration of these bioactives in their natural matrix, has not
heretofore been achieved or recognized. Components of the
bioactive-rich concentrate acts are believed to act synergistically
to produce a more potent and greater hypolipidemic and
hypocholesterolemic effect than the art has recognized, e.g., when
individual, singly purified bioactives are employed. In accordance
with the invention, these natural micronutrients and antioxidants
are captured, extracted and concentrated without disrupting or with
minimal disruption of the natural matrix within which they exist.
Since these micronutrients and antioxidants are in their natural
lipid matrix, they are more bioavailable and potent than in their
purified isolated form.
[0013] One embodiment of the invention features a bioactive-rich
concentrate derived from the unsaponifiable fraction of rice germ
oil, which includes the unsaponifiable content of rice oil. The
concentrate is substantially free of rice bran contaminants and may
be substantially free of oil matter such as free fatty acids, fats,
and triglycerides, and the unsaponifiable content is present at a
concentration 10 to 100 times greater than in raw recovered rice
bran oil or rice germ oil. The unsaponifiable contents typically
include a 4-dimethyl sterol component, a 4-monomethyl sterol
component, a .gamma.-oryzanol component, a polyphenol component, a
tocopherol component, and a tocotrienol component.
[0014] In an embodiment, the amount of 4-dimethyl sterol component
may be about 10-30%, the amount of 4-monomethyl sterol component
may be about 4-14%, the amount of .gamma.-oryzanol component may be
about 20-40%, the amount of polyphenol fraction component may be
about 5-15%, the amount of tocopherol component may be about 5-20%,
and the amount of tocotrienol component may be about 5-20%; by
weight (based on total weight of the concentrate.)
[0015] The bioactive-rich concentrate may be used alone or as part
of a formulation, e.g., in combination with a cholesterol lowering
drug and/or an HMG CoA reductase inhibitor. Such compositions may
be advantageously used for, e.g., lowering serum lipids,
cholesterol, blood glucose, triglycerides, and/or HDL-C levels. In
formulations or combination therapies, the bioactive-rich
concentrate may be present in an amount from about 250 mg to about
1000 mg; the cholesterol lowering drug is present in amount from
about 10 mg to about 50 mg, or from about 10 mg to about 100 mg.
The HMG CoA reductase inhibitor, e.g., mevastatin, lovastatin,
pravastatin, simvastatin, fluvastatin, cerivastatin, atorvastatin,
tenivastatin, rosuvastatin, pitavastatin and combinations thereof,
is generally present in amount from about 1 mg to about 80 mg.
Cholesterol lowering drugs may include bile acid sequestrants such
as cholestyramine, colesevelam and colestipol; or fibric acid
derivatives such as fenofibrate and gemfibrozil.
[0016] Other embodiments of the invention include methods
comprising administering the bioactive-rich concentrate, for
example, orally, to treating a patient in need thereof suffering
from hypertension, hyperlipidemia, obesity, inflammatory disease,
arthritis, hypercholesterolemia, cardiovascular disease,
cerebrovascular disease, arteriosclerosis, diabetes mellitus,
immune dysfunction or cancer, wherein the patient is administered a
bioactive-rich concentrate derived from the unsaponifiable fraction
of rice germ oil, in a effective dosage regime sufficient to treat
the patient.
[0017] Thus, the method may comprise treating a patient in need of
treatment for total serum cholesterol, LDL-C, apolipoprotein B,
triglycerides, improving HDL-C levels, inhibiting platelet
aggregation and dissolving aortic streaks, by administering to the
patient a bioactive-rich concentrate derived from the
unsaponifiable fraction of rice bran oil or rice germ oil, or a
composition comprising such a bioactive rich concentrate, in a
effective dosage regime sufficient to treat the patient.
[0018] Especially useful embodiments of the invention include food
products containing a bioactive-rich concentrate derived from the
unsaponifiable fraction of rice bran oil or rice germ oil, e.g.,
breakfast cereals, snack or energy bars, butter substitutes,
margarines, salad dressings, mayonnaises, or beverages. Food
products of the invention generally contain from about 0.1% to 15%
of the bioactive-rich concentrate.
[0019] In still another aspect, the invention provides methods for
the manufacture of the concentrate disclosed herein comprising
thoroughly separating rice germ component solids from remaining
solids in rice germ or bran, extracting the oil from the rice germ
or bran, and separating the saponifiable from the unsaponifiable
components. The latter step may by accomplished by any known method
that can produce a micronutrient concentrate in its natural matrix,
and preferably is done in the cold immediately after the oil
recovery step. The effect is to minimize damage to the
micronutrients in the unsaponifiable concentrate otherwise induced
by enzymes present in the oil and non germ rice bran components,
and greatly to increase the concentration of these micronutrients,
while maintaining the microcomponents in their natural chemical
state. The concentrate permits the manufacture of convenient dose
forms that can be consumed in a pill or capsule to achieve health
benefits heretofore obtainable only by consuming much larger
quantities of rice bran oil.
BRIEF DESCRIPTION OF THE FIGURES
[0020] FIG. 1 illustrates the steps in the cholesterol homeostasis
pathway.
[0021] FIG. 2 illustrates the performance of a bioactive-rich
concentrate of the invention as compared to a placebo high
cholesterol diet. The comparison is based on a measure of total
cholesterol reduction in hamsters after 9 weeks of administering
the treatment.
[0022] FIG. 3 illustrates the performance of a bioactive-rich
concentrate of the invention as compared to a placebo high
cholesterol diet. The comparison is based on a measure of
LDL-cholesterol reduction in hamsters after 9 weeks of
administering the treatment.
[0023] FIG. 4 illustrates the performance of a bioactive-rich
concentrate of the invention as compared to a placebo high
cholesterol diet. The comparison is based on a measure of
HDL-cholesterol in mg/dL in hamsters after 9 weeks of administering
the treatment.
[0024] FIG. 5 illustrates the performance of a bioactive-rich
concentrate of the invention as compared to a placebo high
cholesterol diet. The comparison is based on a measure of blood
glucose levels in hamsters after 9 weeks of administering the
treatment.
DETAILED DESCRIPTION
Definitions
[0025] For convenience, certain terms used in the specification,
examples, and appended claims are collected here.
[0026] "Phytonutrient" as used herein is includes nonnutritive
bioactive plant substance, such as a flavonoids or carotenoids,
considered to have a beneficial effect on human health.
[0027] "Micronutrients" include phytonutrients, vitamins and
minerals which are essential for healthy body function, as well
components present in, for example, rice germ oil in very small
concentrations that have not been fully characterized but
contribute to the known health effects of rice oil
preparations.
[0028] "Bioactives" collectively include phytonutrients,
micronutrients and antioxidants.
[0029] "Rice bran contaminants" include all components of rice bran
excepting the rice germ component.
[0030] "(Natural) matrix" refers to the proportion of
naturally-occurring components and the form in which the components
are present in rice germ oil. For example, rice oil includes
phytosterols that are present naturally as a range of phytosterols
of specific chemical structures, and particular esters thereof
"Matrix," in this case, refers to the phytosterol fraction as just
described. The natural matrix represents the bioactives occurring
in their native form in their natural lipid environment. A key
concept of the present invention is that the bioactives, e.g., the
micronutrients, in their natural matrix are biologically more
potent than their isolated (purified or separately extracted)
forms. Conventional techniques used by others to isolate these
micronutrients separate them from their lipid substrate and degrade
their molecular structure. For example, phytosterols that are
present in RBO exist as free sterols as well as sterol esters. They
occur in a dissolved state in the natural lipids of RBO. Together,
the micronutrient (in its original form) and the lipid in which it
exists are termed as the natural matrix. The present invention
ensures that the bioactives, e.g., micronutrients, are maintained
as they are in their natural lipid environment.
[0031] Combination therapy" (or "co-therapy") includes the
administration of a bioactive-rich concentrate of the invention and
at least a second agent as part of a specific treatment regimen
intended to provide the beneficial effect from the co-action of
these therapeutic agents. The beneficial effect of the combination
includes, but is not limited to, pharmacokinetic or pharmacodynamic
co-action resulting from the combination of therapeutic agents.
Administration of these therapeutic agents in combination typically
is carried out over a defined time period (usually minutes, hours,
days or weeks depending upon the combination selected).
"Combination therapy" may, but generally is not, intended to
encompass the administration of two or more of these therapeutic
agents as part of separate monotherapy regimens that incidentally
and arbitrarily result in the combinations of the present
invention. "Combination therapy" is intended to embrace
administration of these therapeutic agents in a sequential manner,
that is, wherein each therapeutic agent is administered at a
different time, as well as administration of these therapeutic
agents, or at least two of the therapeutic agents, in a
substantially simultaneous manner. Substantially simultaneous
administration can be accomplished, for example, by administering
to the subject a single capsule having a fixed ratio of each
therapeutic agent or in multiple, single capsules for each of the
therapeutic agents. Sequential or substantially simultaneous
administration of each therapeutic agent can be effected by any
appropriate route including, but not limited to, oral routes,
intravenous routes, intramuscular routes, and direct absorption
through mucous membrane tissues. The therapeutic agents can be
administered by the same route or by different routes. For example,
a first therapeutic agent of the combination selected may be
administered by intravenous injection while the other therapeutic
agents of the combination may be administered orally.
Alternatively, for example, all therapeutic agents may be
administered orally or all therapeutic agents may be administered
by intravenous injection. The sequence in which the therapeutic
agents are administered is not narrowly critical. "Combination
therapy" also can embrace the administration of the therapeutic
agents as described above in further combination with other
biologically active ingredients and non-drug therapies (e.g.,
surgery or radiation treatment.) Where the combination therapy
further comprises a non-drug treatment, the non-drug treatment may
be conducted at any suitable time so long as a beneficial effect
from the co-action of the combination of the therapeutic agents and
non-drug treatment is achieved. For example, in appropriate cases,
the beneficial effect is still achieved when the non-drug treatment
is temporally removed from the administration of the therapeutic
agents, perhaps by days or even weeks.
[0032] One embodiment of the invention features a bioactive-rich
concentrate derived from the unsaponifiable fraction of rice bran
oil or rice germ oil, which includes the unsaponifiable content of
rice oil; the concentrate is substantially free of rice bran
contaminants; and the unsaponifiable content is present at a
concentration 10 to 100 times greater than in rice bran oil or rice
germ oil. The unsaponifiable contents include a 4-dimethyl sterol
component, a 4-monomethyl sterol component, a .gamma.-oryzanol
component, a polyphenol component, a tocopherol component, and a
tocotrienol component.
[0033] In an embodiment, the amount of 4-dimethyl sterol component
may be 10-30%, the amount of 4-monomethyl sterol component may be
4-14%, the amount of .gamma.-oryzanol component may be 20-40%, the
amount of polyphenol component may be 5-15%, the amount of
tocopherol component may be 5-20%, and the amount of tocotrienol
component may be 5-20%; by weight (based on total weight of the
concentrate.)
[0034] The bioactive-rich concentrate may be used alone or as part
of a formulation, e.g., in combination with a cholesterol lowering
drug and/or an HMG CoA reductase inhibitor. Such compositions may
be advantageously used for, e.g., lowering serum lipids,
cholesterol, blood glucose, triglycerides, and/or HDL-C levels. In
formulations or combination therapies, the bioactive-rich
concentrate may be present in an amount from about 250 mg to about
1000 mg; the cholesterol lowering drug is present in amount from
about 250 mg to about 1000 mg. The HMG CoA reductase inhibitor,
e.g., mevastatin, lovastatin, pravastatin, simvastatin,
fluvastatin, cerivastatin, atorvastatin, tenivastatin,
rosuvastatin, pitavastatin and combinations thereof, is generally
present in amount from about 1 mg to about 80 mg. Cholesterol
lowering drugs may include bile acid sequestrants such as
cholestyramine, colesevelam and colestipol; or fibric acid
derivatives such as fenofibrate and gemfibrozil.
[0035] Other combination compositions contemplated herein include a
composition that includes a bioactive-rich concentrate and natural
components such as herbs and/or herbal ingredients. Such herbs
and/or herbal ingredients may include those used to lower
cholesterol levels, such as guggul gum resin derived from or in the
form of gum from Commiphora mukul, policosanol that may be derived
from sugar cane wax, curcumin that may be derived from turmeric,
garlic, psyllium, green tea and/or green tea extracts, and licorice
and/or licorice extracts. Other combination compositions can
include such ingredients as DHA, EPA, CoQ10, fish oil, olive oil,
vitamin E and vitamin C.
[0036] Especially useful embodiments of the invention include food
products containing a bioactive-rich concentrate derived from the
unsaponifiable fraction of rice bran oil or rice germ oil. Such
food products may include, for example, breakfast cereals, snack or
energy bars, butter substitutes, margarines, salad dressings,
mayonnaises, or beverages. Food products of the invention generally
contain from about 0.1% to about 15% of the bioactive-rich
concentrate.
[0037] Pure rice germ may be used as one preferred source of oil
from which the bioactive-rich concentrates of the invention may be
obtained. Rice germ is a by-product of the rice milling industry
and may comprise a portion of rice bran. When the lipases present
in the germ come into contact with the oil during milling, they can
hydrolyze the triglycerides into free fatty acids, which damage the
micronutrients. Preferably, the rice germ is polished repeatedly,
for example polished with three or four passes or more, and
delicately. Hullers should be avoided, and the mills should
separate the bran generated from the polishes as well as
stringently separate stone and grit from the paddy. Raw rice
milling may be more desirable. Rice bran should be separated from
the germ after the initial polishing by sieving and air
classification, or other density based separation technique, more
desirably within 24 hours after polishing, followed by cold
storage, e.g., 3-5.degree. C., to stabilize the germ and halt any
residual lipase activity.
[0038] The germ should be processed as soon as possible after cold
storage is initiated. The rice germ oil may be extracted from the
germ by cold pressing; solvent extraction, e.g., with non-polar
solvents such as hexane, or polar solvents such ethanol or
isopropanol; or supercritical fluid extraction techniques, e.g.,
super critical fluid fractionation, CO.sub.2, fluorohydrocarbons,
or propane. Such extraction techniques are known by those of skill
in the art.
[0039] Alternately, the bioactive-rich concentrates of the
invention may be obtained from rice bran oil deodorizer distillate
(DOD). DOD is obtained as a by-product of the rice bran oil
refining industry. Deodorization is the final step of the rice oil
refining process; the deodorization step removes the unwanted
impurities in crude rice bran oil, such as aldehydes, ketones and
sulfur compounds. This process consists of heating the degummed,
deacidified, and bleached rice bran oil to about 250.degree. C.
under high vacuum and steam injection. During this process, several
key micronutrients in rice bran oil such as tocotrienols and
tocopherols can be distilled away with the impurities. There is
also a significant amount of free fatty acids (FFAs, up to about
60%) in rice bran oil DOD. As such, rice bran oil DOD is less
desirable than rice germ oil, but may be used in the invention
under controlled conditions. DOD for use in the invention should
employ minimal alkali treatment during de-acidification, so that
the micronutrients in the oil are preserved. Further, it should be
used within a short time, e.g., one week, of its creation, to
minimize FFA micronutrient degradation.
[0040] The bioactive-rich concentrates of the invention may be
produced from crude rice germ oil by, for example any method that
would yield the heretofore unobtained high concentration of the
components from the unsaponifiable fraction components. Such
methods include preferential concentration (selective solubility)
using polar solvents such as ethyl alcohol, isopropyl alcohol,
etc.; fractionation of the bioactives of the germ oil after
converting the FFAs present in the germ oil to methyl esters
followed by SCF treatment; and/or FFA conversion to the ethyl or
methyl esters using conventional techniques, followed by short path
distillation.
[0041] These techniques, which are familiar to those in the art,
yield a product, which is 5 to 10 times higher in concentration of
the key bioactives (e.g., tocotrienols, tocopherols,
.gamma.-oryzanol, phytosterols, polyphenols, etc.)
[0042] If DOD is used as a raw material, the following techniques
may be used to obtain a product with a high concentration of
unsaponifiable fraction components: neutralization of FFAs present
in the germ oil with sodium bicarbonate, sodium carbonate, sodium
hydroxide or calcium hydroxide, followed by filtration of the FFA
salts; or converting the FFAs into its (m)ethyl ester, followed by
short path distillation or super critical fluid treatment such as
super critical fluid fractionation.
[0043] These techniques, which are familiar to those in the art,
yield the product of the invention, which is about 10, about 20,
about 30, about 40 or about 50 to 100 times greater in
concentration of certain key micronutrients, such as tocopherols
and tocotrienols.
[0044] As a reference, the composition of the unsaponifiable
content of crude RBO is shown below:
TABLE-US-00003 Constituent Concentration Total Unsaponifiable
Matter 4.2% 4-dimethyl sterols 1.8% 4-monomethyl sterols 0.4%
.gamma.-oryzanol 1.2% Polyphenols and hydrocarbons 0.8% (squalene,
.gamma.-amino butyric acid) Tocopherols 0.04% Tocotrienols 0.07%
Ref: Uniliver paper on "Minor constituents of RBO by Deckere and
Kover, Nutrition Reviews 54(11) S120-S126
The relative concentration of the phytosterol components of RBO are
campasterol: 0.506 (mg/100 g); stigmasterol: 0.271 (mg/100 g);
betasitosterol: 0.885 (mg/100 g); other sterols: 0.3 (mg/100
g).
[0045] The compositions of the invention (i.e., after processing as
described hereinabove) are characterized as follows:
TABLE-US-00004 Constituent Concentration Total Unsaponifiable
Matter 25-70% 4-dimethyl sterols (phytosterols) 10-30% 4-monomethyl
sterols 4 to 14% .gamma.-oryzanol 20 to 40% Polyphenols and
hydrocarbons (e.g., 5 to 15% squalene, gamma-amino butyric acid)
Tocopherols (e.g., including .alpha.-, .beta.-, .gamma.- and 5 to
20% .delta.-isomers) Tocotrienols (e.g., including .alpha.-,
.beta.-, .gamma.- and 5 to 25% .delta.-isomers)
[0046] It should be noted that within each class of micronutrient
(tocotrienols, tocopherols, phytosterols, oryzanol, polyphenols),
the relative concentration of individual isomers and components is
the same as the original source (either rice bran/germ). This
"signature" cannot be obtained by mixing isolated purified
components.
[0047] The bioactive-rich concentrates of the present invention may
be produced at an estimated cost of $0.02 to $0.03 per serving,
which is several times lower than the individual pharmaceutical
compounds targeted at the same ailments. As an example,
tocotrienols retail for around $200/kg, and .gamma.-oryzanol
retails for around $75/kg.
[0048] The preparation of pharmaceutical or pharmacological
compositions containing the compositions of matter of the invention
will be known to those of skill in the art in light of the present
disclosure. Typically, such compositions may be prepared as orally
ingestible preparations such as tablets or other solids for oral
administration. Alternatively, they may be formulated as
injectables, either as liquid solutions or suspensions; solid forms
suitable for solution in, or suspension in, liquid prior to
injection; as time release capsules; or in any other form currently
used, including creams, lotions, mouthwashes, inhalants and the
like.
[0049] While the compositions of the invention may take many forms,
such as capsules, pills and gelcaps, for personal daily or other
intermittent use, they also are especially useful when formulated
as food additives, e.g., in spreads, frozen desserts, beverages and
nutritional bars.
[0050] Examples of food products comprising the compositions of the
invention include margarines or other spreads of oil based
products, bakery products, dairy products, e.g., yogurt, cheese and
milk-based drinks, beverages, e.g., soft drinks, fruit juices and
tea- and coffee-based drinks, sauces, dressings and mayonnaise and
confectionery products, e.g., frozen confectionery products such as
water-ice or ice-cream, and dietary supplements such as health
bars.
[0051] The compounds described herein can be used in compositions
comprising fat and non-fat components to provide general health
benefits, including cardiovascular benefits, such as lowering
cholesterol in the consumer, treating, preventing, and/or
inhibiting heart disease (e.g., atherosclerosis, restenosis,
thrombosis) and treating other conditions such as hypertension,
poor circulation, complications associated with diabetes,
cerebrovascular disease, neurological disorders, and liver
abnormalities.
[0052] The compounds can be used in the production of baked goods
in any form, such as mixes, shelf-stable baked goods (including
health bars), and frozen baked goods. Applications include cakes,
brownies, muffins, bar cookies, health bars, wafers, biscuits,
pastries, pies, pie crusts, and cookies, including sandwich cookies
and chocolate chip cookies. The baked goods can contain fruit,
cream, or other fillings. Other baked good uses include breads and
rolls, crackers, pretzels, pancakes, waffles, ice cream cones and
cups, yeast-raised baked goods, pizzas and pizza crusts, and other
baked salted snacks.
[0053] The compositions herein can be used alone or in combination
with fats to make shortening and oil products. The fats can be
synthetic or derived from animal or vegetable sources, or
combinations of these. Shortening and oil products include
shortenings, margarines, spreads, butter blends, lards, cooking and
frying oils, salad oils, popcorn oils, salad dressings, mayonnaise,
and other edible oil products. In a particular embodiment of the
present invention, the compositions include margarines, butter,
dressings and spreads.
[0054] Other uses for the compositions of the present invention
include partial or complete replacement fats and/or oils present in
peanut butter, frozen desserts such as ice cream and ice cream
coatings, whipped toppings, frosting products, processed meat
products such as vegetable protein-based meat analog products,
sauces, gravies, and dairy products such as milkshakes, milk
products, coffee whiteners, and cheese products.
[0055] The compounds described herein may also be used in beverage
compositions, e.g., dilute water beverages (also called
"near-water" beverages), milks, coffees, teas, colas, and fruit
juices.
[0056] Compositions of the invention may be used as the
pharmaceutical, nutraceutical, cosmeceutical and health food
dietary supplements for treating health disorders including high
blood pressure, hypercholesterolemia, hyperlipidemia,
cardiovascular disease, cerebrovascular disease, diabetes, cancer,
obesity, inflammatory diseases, arthritis, improve immune function;
as a sports and weight loss formulations in improving lean body
mass and liver disorders; and other uses including skin care, hair
growth, UV protection, antidandruff agents, and cosmeceuticals.
EXEMPLIFICATION
[0057] The invention now being generally described, it will be more
readily understood by reference to the following examples which are
included merely for purposes of illustration of certain aspects and
embodiments of the present invention, and are not intended to limit
the invention in any way.
[0058] Composition A:
[0059] Composition A is derived from two starting materials: rice
germ and deodorizer distillate. Purified and stabilized rice germ
from the bran is obtained from the first two polishes of rice using
2 successive steps. First, size based separation or sieving is
conducted. Density based separation (air classification) is then
performed. These operations are carried out within 24 Hrs of
polishing operation at the mill. These successive steps yield pure
rice germ (over 90% purity). This rice germ is maintained in a cold
storage facility at 3 to 5 degrees Celsius to arrest the lipase
activity and keep it stable. A crude rice germ oil is extracted
from this pure rice germ by solvent extraction with hexane. Hexane
is then removed to leave behind crude rice germ oil. This crude
rice germ oil is then subject to a degumming process, a dewaxing
process, and then esterified. This esterified oil is subject to
short path distillation and a concentrated fraction of the rich
germ oil is collected.
[0060] Rice bran oil deodorizer distillate (DOD) is obtained as a
by-product of the rice bran oil refining industry. This distillate
is subjected to short path distillation and a concentrated fraction
is collected.
[0061] The two collected fractions from rice germ and DOD are
analyzed for their micronutrient content and on the basis of the
analysis are combined in proportions to yield Composition A.
[0062] Composition A is characterized by HPLC and GC. Within each
class of nutrient, the relative concentration of individual isomers
and components is the same as the original source (rice
bran/germ).
[0063] The table below describes a sample preparation of a
composition ("Composition A") made in accordance with the
invention, and which was tested in an animal study described
below:
TABLE-US-00005 Ri-Active .TM. Components g/100 g .gamma.-oryzanol
30 g/100 g Tocopherols (.alpha., .beta., .gamma., d isomers) 4
g/100 g Tocotrienols (.alpha., .beta., .gamma., d isomers) 7 g/100
g Phytosterols (.beta.-sitosterol, sitosterol, campestrol, 7 g/100
g stigmasterol and 27 other sterol derivatives Polyphenols (trans
ferulic acid, protocatechuic acid, 1.0 g/100 g epicatechin,
p-coumaric acid, sinapic acid, tricin, a flavonol, GABA and 8 other
polyphenols) Squalene and other hydrocarbons Trace amounts
[0064] Hypolipidemia, Hypocholesterolemia and Hypoglycemia Study in
Syrian Hamsters
[0065] This study was undertaken to evaluate the efficacy and
safety of a composition of the invention noted above. Syrian golden
hamsters (60) were fed semi-purified diets containing coconut oil
and 0.5% cholesterol to induce moderate hypercholesterolemia and
divided into 6 groups of 10 animals each. Group 1 (Placebo)
continued to be fed the same control diet. Group 2 (Positive
control) was fed 0.1% phytosterols in the control diet (FDA
approved natural hypocholesterolemic agent) Group 3 was fed 0.5%
phytosterols in the control diet, Group 4 was fed 0.1%
Ri-Active.TM. in the control diet, Group 5 was fed 0.5%
Ri-Active.TM. in the control diet, and Group 6 was fed 1%
Composition A in the control diet. All diets were prepared by the
Research Diets, Pennsylvania USA. All animals were fed the
corresponding diets for 9 weeks. Weekly body weights were
monitored. Physical symptoms, if any and gastrointestinal function
were also monitored in all the animals. Blood was drawn by retro
orbital sinus at the end of 4, 6 and 9 weeks, from each animal and
analyzed for total cholesterol, LDL-C, Triglycerides, HDL-C, serum
glucose levels AST and ALT.
[0066] Table 3 shows results of the Watson clinical study,
investigating the effects of RBO performed using a RBO
unsaponifiable fraction, using rice bran oil as a whole. This study
was conducted on 50 human subjects over a 12 month period showed
that administering 3.1 g/day of RBO unsaponifiables resulted in a
14.1% reduction in total cholesterol and a 20.5% reduction in LDL-C
in addition to an increase in HDL-C levels and significant decrease
in triglycerides levels
TABLE-US-00006 TABLE 3 Effect of RBO unsaponifiable fraction in
hypercholesterolemic subjects Methodology Lipid Parameters
Reduction/Increase Reference 50 hypercholesterolemic subjects Total
Cholesterol -14.1% (p < 0.05) T. R. Watkin, M. Geller, D. K.
Kooyenga received in random blinded LDL-Cholesterol -20.6% (p <
0.05) and M. L. Bierenbaum; fashion 3.1 g of RBO non- HDL-C/Total
Cholesterol +41.17% (p < 0.025) Environmental saponifiables or
placebo for 12 Triglycerides/HDL-C -43.98% (p < 0.05) and
Nutritional Interactions, months 3: 115-122, 1999
[0067] The current studies were undertaken at least in part to
investigate if Composition A, with ten times the concentration of
that used in Watkin's study, and having multiple bioactives from
the RBO unsaponifiable fraction would act synergistically resulting
in the overall cardiovascular risk benefits and would lower LDL
cholesterol to an even greater extent while increasing the HDL-C
levels.
[0068] Test Results: (Comparison Between Groups)
[0069] The results indicated significant hypocholesterolemic,
hypolipidemic and hypoglycemic effect of Composition A compared to
two groups: 1) phytosterols, which is the positive control or "gold
standard", and are currently the only natural products in the
market with a FDA health claim for cardiovascular health (at least
800 mg of phytosterols or 1300 mg of phytosterol esters in two
meals per day); and 2) a placebo control (no treatment) group.
Phytosterols may compete with cholesterol uptake releasing
cholesterol for excretion. The average decline in LDL-c levels
through the use of phytosterols is thought to be between 8%-15% in
humans.
[0070] 0.1% Composition A (.about.10 mg dosage):
[0071] In the 0.1% Composition A group, total cholesterol levels
were 16% lower (242 mg/dl) and statistically significant
(P<0.008) when compared to placebo group (287 mg/dl) and more
than 6% greater reduction than the 0.1% phytosterols group (260
mg/dl). The significance of this result is further underscored
because the 0.1% Composition A group contains less than 1/10th the
concentration of phytosterols as compared to the 0.1% phytosterol
group. FIG. 2 shows a comparative performance of Composition A as
measured by total cholesterol reduction.
[0072] The decrease in LDL-C in this group (135 mg/dl) was
remarkable and significant (P<0.001) with 22% reduction when
compared to placebo group (171 mg/dl) and nearly 8% greater
reduction when compared with 0.1% phytosterol group (146 mg/dl) as
seen in FIG. 3.
[0073] Triglycerides also showed an excellent reduction of 21% (242
mg/dl) when compared with placebo (306 mg/dl) and this was 12%
greater to reduction shown by the 0.1% phytosterol group (279
mg/dl).
[0074] 0.5% Composition A (.about.50 mg dosage):
[0075] Increasing Composition A dosage by 5 times showed a
significant increase in HDL-C levels. HDL-C levels in this group
(127 mg/dl) increased by 10% more than the placebo group (115
mg/dl) and 17.6% more than the 0.5% phytosterol group (108 mg/dl).
These results were statistically significant with P<0.022 (FIG.
4). This indicates a natural product that demonstrates an increase
in HDL-C along with a significant decrease in LDL-C. This finding
may be supported by previously published human clinical studies
conducted over a 12-month period with RBO unsaponifiable fraction
(Watkins et al. in 1990: Table 3). The phytosterol groups showed a
reduction in HDL-C levels as predicted by previous studies.
[0076] Increasing Composition A dosage by 5 times also showed
significant hypoglycemic effect (P<0.015) by lowering blood
glucose levels to 93 mg/dl which was 23% lower compared to the
placebo diet (121 mg/dl) and 15.6% reduction compared to the 0.5%
phytosterol treatment group (111 mg/dl). These results were
statistically significant with P<0.030 (See FIG. 5).
[0077] Increasing the phytosterol dosage by 5 times (0.5%
phytosterol group) did not show any statistically significant
reduction in either total cholesterol or LDL-C as compared to the
0.1% Composition A group (0.1% Composition A group had 1/50th the
concentration of phytosterols as compared to the 0.5% phytosterol
group). Further, there was no statistical significance in the total
cholesterol, LDL-C and Triglycerides reduction between the 0.5%
phytosterol group and 0.5% Composition A groups.
[0078] Animals as their Own Control:
[0079] The group comparisons above indicate a statistically
significant improvement in lipid parameters for the 0.1%
Composition A group, the 0.5% Composition A group and the 0.5%
phytosterol group (0.5% phytosterol group is superior to 0.1%
phytosterol group as predicted by prior studies).
TABLE-US-00007 TABLE 6 Lipid parameters of Placebo group Parameter
Peak (Week 4) Final (Week 9) Change(%) Total Cholesterol 396 .+-.
87 287 .+-. 26 -27.5 LDL-C 272 .+-. 77 171 .+-. 18 -37.1 HDL-C 124
.+-. 30 115 .+-. 15 -7.2 Triglycerides 470 .+-. 220 306 .+-. 111
-34.9 Glucose 125 .+-. 28 121 .+-. 15 -3.2
TABLE-US-00008 TABLE 7 Lipid parameters of 0.1% Ri-Active Peak
Final Change Parameter (Week 4) (Week 9) (%) Total Cholesterol 381
.+-. 141 243 .+-. 36 -36.2 LDL-C 271 .+-. 166 135 .+-. 21 -50.1
HDL-C 110 .+-. 20 108 .+-. 20 N.C. Triglycerides 379 .+-. 286 242
.+-. 69 -36.1 Glucose 126 .+-. 67 110 .+-. 17 -12.8
TABLE-US-00009 TABLE 8 Lipid parameters of 0.5% Ri-Active Peak
Final Change Parameter (Week 4) (Week 9) (%) Total Cholesterol 436
.+-. 126 279 .+-. 61 -36 LDL-C 322 .+-. 109 153 .+-. 41 -52.4 HDL-C
115 .+-. 24 127 .+-. 21 10.3 Triglycerides 532 .+-. 224 291 .+-.
148 -54.7 Glucose 121 .+-. 42 93 .+-. 8 -22.7
TABLE-US-00010 TABLE 9 Lipid parameters of 0.5% Phytosterols group
Peak Final Change Parameter (Week 4) (Week 9) (%) Total Cholesterol
340 .+-. 143 279 .+-. 26 -18 LDL-C 233 .+-. 134 134 .+-. 18 -42.5
HDL-C 108 .+-. 16 108 .+-. 11 N.C. Triglycerides 440 .+-. 346 259
.+-. 126 -41.1 Glucose 122 .+-. 47 111 .+-. 21 -9.5
TABLE-US-00011 TABLE 10 Ratio Comparison of Treatment Groups
TC/HDL-C LDL-C/HDL-C Treatment Initial Final Initial Final Placebo
3.28 2.51 2.28 1.51 0.1% Ri-Active 3.40 2.25 2.45 1.25 0.5%
Ri-Active 3.78 2.19 2.79 1.20 0.5% Phytosterols 3.15 2.58 2.16
1.34
[0080] Safety
[0081] Composition A showed good animal growth performance and
health maintenance at very high doses. It did not show any adverse
effects or any physical signs and symptoms of toxicity. Further,
the AST and ALT measurement made by increasing the Composition A
dosage by 10 times (1% Composition A) were not different from the
control group and indicated its safety.
[0082] Observations
[0083] 1. Composition A at 0.1% (10 mg Composition A in the diet)
contains 1.1 mg Tocos (Tocopherol and Tocotrienols), 3.0 mg Gamma
Oryzanol and less than 1.0 mg of phytosterols. In the light of
these low bioactive concentrations, the observed significant
hypolipidemic effect in 0.1% Composition A group is significant. It
shows the synergistic effect of RBO bioactives acting in concert at
very low concentration in their natural matrix. Previous studies on
individually purified RBO bioactives have never shown any
hypocholesterolemic or hypolipidemic at such low
concentrations.
[0084] 2. Composition A at 5 times the concentration (0.5%)
demonstrated a significant increase in HDL-C, which is a positive
factor to reduce the risk of CVD.
[0085] 3. Composition A at 5 times the concentration (0.5%)
demonstrated a significant hypoglycemic effect, which is again risk
factor of cardiovascular disease.
[0086] 4. This study demonstrates that Composition A is safe and
superior to plant phytosterols (the gold standard) for the
reduction of CVD risk factors. These results indicate significant
hypocholesterolemic, hypolipidemic and hypoglycemic effect of 0.1%
Composition A compared with 0.1% phytosterols which may be a "gold"
standard, as well as compared with placebos. Total cholesterol
levels were lower and statistically significant (P value between
groups is 0.010). It has shown a reduction of 16% compared to
placebo; and more than 6% reduction compared to phytosterols @
1/10th the sterol concentration, which is statistically significant
(P value between groups less than 0.001). The decrease in LDL-C was
22% when compared with placebo; and nearly 8% decrease when
compared with 0.1% phytosterols @ 1/10th the sterol concentration.
Increasing Composition A dosage by 5 times (0.5% Composition A
group) showed a significant increase of HDL-C levels by 10% over
the placebo (high cholesterol diet) group. This natural product
demonstrates an increase in HDL-C, which is a well-recognized
positive factor in controlling CVD. Increasing Composition A dosage
by 5 times (0.5% Composition A group) also showed significant
hypoglycemic effects by lowering blood glucose levels by 23% over
the placebo group. Composition A showed good growth performance and
did not show any adverse effects by any signs and symptoms or by
the AST and ALT measurement which were not different from the
control group indicating its safety.
[0087] The results shown in the above examples suggest that:
[0088] 1. 0.1% Composition A (10 mg of Composition A in the diet)
includes 1.1 mg Tocos, 3.0 mg Gamma Oryzanol and less than 1.0 mg
of phytosterols. The observed results show a significant
synergistic effect of RBO bioactives acting in concert at very low
concentration in their natural matrix.
[0089] 2. Composition A at 5 times the concentration (of 0.1%,),
demonstrate significant raise in HDL-C, which is a positive factor
to reduce the risk of CVD.
[0090] 3. Composition A at 5 times the concentration (of 0.1%)
demonstrated significant hypoglycemic effect, which is a factor to
reduce the risk of cardiovascular disease..
[0091] 4. Composition A at 5 times the concentration (of 0.1%),
demonstrate significant raise in HDL-C, which is a positive factor
to reduce the risk of CVD.
[0092] 5. Composition A at 5 times the concentration (of 0.1%)
demonstrated significant hypoglycemic effect, which is again a
factor to reduce the risk of cardiovascular disease.
[0093] 6. Composition A demonstrated to be safe without any adverse
effects and more efficacious than a "gold" standard,
phytosterol.
EQUIVALENTS
[0094] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of the invention
and are covered by the following claims. Various substitutions,
alterations, and modifications may be made to the invention without
departing from the spirit and scope of the invention as defined by
the claims. Other aspects, advantages, and modifications are within
the scope of the invention. The contents of all references, issued
patents, and published patent applications cited throughout this
application are hereby incorporated by reference. The appropriate
components, processes, and methods of those patents, applications
and other documents may be selected for the invention and
embodiments thereof.
[0095] Also incorporated by reference are the following: Nicolosi
R. J., Ausman L. M., and Hegstead M. (1991) "Rice bran oil lowers
serum and LDL lipoprotein cholesterol and apo-B levels in non-human
primates" Atherosclerosis. 88(2-3) 133-142; Rukmini C., and
Raghuram T. C., (1991) "Nutritional and biochemical aspects of the
hypolipidemic action of rice bran oil: A review" J. Amer. Coll.
Nutrition. 10: 366-375; Sugano M., and Tsuji E., (1997) "Rice bran
oil and cholesterol metabolism". J. of. Nutrition, 127(3):
521S-524S; Qureshi A. A., Qureshi N., Hasler-Rapacz J. O., (1991)
"Dietary tocotrienols reduce concentrations of plasma cholesterol,
apolipoprotein B, thromboxane B2 and platelet factor 4 in pigs with
inherited hyperlipidemias." Am. J. Clin. Nutr. 53:1021S; Qureshi A.
A., Bradlow B. A., Salser W. A., Brace L. D., (1997), "Novel
tocotrienols of rice bran modulate cardiovascular disease risk
parameters of hypercholesterolemic humans," J. Nutr. Biochem 8:
1-9; Nesaratnam K., Stephen R., Dils R., Darbre P., (1998)
"Tocotrienols inhibit the growth of breast cancer cells
irrespective of estrogen receptor status." Lipids, 33(5): 461-469;
Tomeo A. C., Geller M., Watkins T. R., (1995) "Antioxidant effects
of tocotrienols in patients with hyperlipidemia and carotid
stress." Lipids, 30:1179; Xu Z, Hua N., Godber J. S., (2001),
"Antioxidant activity of tocopherols, tocotrienols, and gamma
oryzanol components from rice bran against cholesterol oxidation
accelerated by 2,21-azobis(2-methylpropionamideine)
dihydrochloride." J. Agric. Food. Chem, 49(4): 2077-81.; Ando Y.,
Pharcos I., (1984) "Effect of gamma oryzanol as sunscreen agent",
Jpn. J. Cosmetic Sci. Soc, (1) 41; Akihisa T., Yasukawa K., Yamaura
M., Ukiya M., Kimura Y., Shimizu N., and Arai K., (2000)
"Triterpene alcohol and sterol ferulates from rice bran and their
antiinflammatory effects", J. Agri. Food. Chem, 48:2313-2319; Ieiri
T., Kase N., Hashigami Y., Nakumura T., Shimoda S., (1982) "Effect
of gamma oryzanol on the hypothalamo-pituitary axis in the rat",
Nippon Naibunpi Gokkai Zasshi 58(10):1350-6; Bouic P. J. D.,
Etsbeth S., Liebenberg R. W., Albrecht C. F, Pegel G., Van
Jaarsveld P. P., (1996) "Beta-sitosterol and beta-sitosterol
glucoside stimulate human peripheral blood lymphocyte
proliferation: Implication for their use as an immunomodulatory
vitamin combination", International. J. Immunopharmacology, 18(12):
693-700; Ann Hudson, Dinh P. A., Kokubun T., Simmonds S. J., and
Gescher A., (2000) "Characterization of potentially chemopreventive
phenols in extracts of brown rice that inhibit the growth of human
breast and colon cancer cells". Cancer Epidemiology, Biomarkers
& Prevention. 9: 1163-1170; Sunita T. Manorama R., and Rukmini
C. (1996)"Lipid profile on blended oils with rice bran oil--A study
in human subjects". Report to Dabur India Ltd. Asia Pacific Journal
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[0096] Commonly-owned patent application U.S. Ser. No. 11/251,875,
which claims priority to provisional patent application 60/619,879
filed Oct. 18, 2004 are both hereby incorporated by reference.
* * * * *