U.S. patent application number 09/783884 was filed with the patent office on 2001-11-29 for anticholesterolemic edible oil.
Invention is credited to Berry, Christopher J., Bierenbaum, Marvin L..
Application Number | 20010046548 09/783884 |
Document ID | / |
Family ID | 27359040 |
Filed Date | 2001-11-29 |
United States Patent
Application |
20010046548 |
Kind Code |
A1 |
Berry, Christopher J. ; et
al. |
November 29, 2001 |
Anticholesterolemic edible oil
Abstract
An edible oil is provided that decreases the synthesis,
absorption and blood level of cholesterol by a human patient,
increases the excretion of cholesterol from the human patient,
curtails accumulation of peroxidized material in the blood of the
human patient, and also increases the blood level of vitamin E in
said human patient.
Inventors: |
Berry, Christopher J.;
(Phuket, TH) ; Bierenbaum, Marvin L.; (Montclair,
NJ) |
Correspondence
Address: |
ROD S. BERMAN, ESQ.
JEFFER, MANGELS, BUTLER & MARMARO LLP
Tenth Floor
2121 Avenue of the Stars
Los Angeles
CA
90067
US
|
Family ID: |
27359040 |
Appl. No.: |
09/783884 |
Filed: |
February 14, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09783884 |
Feb 14, 2001 |
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09411591 |
Oct 4, 1999 |
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6277431 |
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60104227 |
Oct 14, 1998 |
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Current U.S.
Class: |
426/601 ;
514/182 |
Current CPC
Class: |
A23L 33/11 20160801;
A23L 33/115 20160801; A23D 9/007 20130101 |
Class at
Publication: |
426/601 ;
514/182 |
International
Class: |
A23D 007/005; A23D
009/007 |
Claims
We claim:
1. An edible oil that reduces the synthesis, absorption and blood
level of cholesterol by a human patient and increases the excretion
of cholesterol from said human patient.
2. The edible oil of claim 1 which comprises at least one compound
that reduces cholesterol absorption in said human patient.
3. The edible oil of claim 1 which comprises at least one compound
that reduces cholesterol synthesis by said human patient.
4. The edible oil of claim 1 which comprises at least one compound
that increases cholesterol excretion from said human patient.
5. The edible oil of claim 1 which is substantially free of trans
fatty acids.
6. The edible oil of claim 1 which attenuates the blood level of
peroxides.
7. The edible oil of claim 1 which provides at least one compound
in the oil that limits the formation and accumulation of TBARS in
hypercholesterolemic human subjects.
8. The edible oil of claim 1 which provides increased levels of
tocopherol or tocotrienol antioxidant activity to the blood, and
other tissues, of human subjects.
9. The edible oil of claim 1 which significantly decreases the
blood level of total cholesterol, LDL cholesterol and
triglyceride/HDL cholesterol in a hypercholesterolemic human
subject, with blood cholesterol in excess of 5.6 .mu.mol./L.
10. The edible oil of claim 1 which significantly increases the
blood level of HDL cholesterol, expresses as the ratio of blood HDL
cholesterol/total cholesterol, in hypercholesterolemic
subjects.
11. The edible oil of claim 1 which lowers the blood level of
peroxides, measured as TBARS, in hypercholesterolemic subjects.
12. The edible oil of claim 1 which significantly raises the blood
level of vitamin E activity in hypercholesterolemic human
subjects.
13. A food product comprising the oil of claim 1.
14. The food product of claim 13 which is selected from the group
consisting of butter, margarine, ice cream and mayonnaise.
15. The food product of claim 13 which is a chocolate product.
16. The food product of claim 13 which is a liquid.
17. The food product of claim 13 which is selected from the group
consisting of milk, soybean milk and rice milk.
18. The food product of claim 13 which is a water-based drink.
19. The food product of claim 13 which is selected from the group
consisting of wines and mineral waters.
20. The edible oil of claim 1 which is a vegetable oil or mixture
of vegetable oils.
21. The edible oil of claim 1 which is a refined rice bran oil.
22. The edible oil of claim 1 which is a mixture of rice bran oil
and a palm oil.
23. A soft gel including a fill comprising the edible oil of claim
1.
24. An edible oil comprising i) about 10 to 30% of tocopherols,
tocotrienols or combinations thereof, ii) about 2 to 20% of free
sterols; iii) about 2 to 20% of sterol esters; iv) about 0.1 to
1.0% of cycloartenols; and, v) about 7 to 19% of saturated fats,
wherein all percentages are weight/weight.
25. The edible oil of claim 24 comprising about 70 to 80% of total
fats.
26. The edible oil of claim 24 comprising less than 2% trans fatty
acids.
27. The edible oil of claim 24 comprising about 20 to 60% of
non-saponifiable components.
28. The edible oil of claim 24 which is a vegetable oil or a
mixture of vegetable oils.
29. The edible oil of claim 24 which is a refined rice bran
oil.
30. The edible oil of claim 24 which is a mixture of a rice bran
oil and a palm oil.
31. A food product comprising the oil of claim 24.
32. An edible oil comprising about 20 to 60% of non-saponifiable
components, said non-saponifiable components comprising (i) at
least one tocotrienol or tocopherol, (ii) at least one free sterol
or sterol ester, and iii) at least one cycloartenol, wherein the
ratio of the amounts of said components i): (ii): (iii) is from
about 1:0.5:0.05 to about 1:5:0.05.
33. The oil of claim 33 wherein said ratio is about 1:1:0.05.
34. A method of reducing total serum cholesterol and serum LDL
cholesterol and raising serum HDL cholesterol in a human patient
comprising the step of administering to said patient an effective
amount of the edible oil of claim 1.
35. The method of claim 34 which also reduces the blood
triglyceride level in a human patient.
36. The method of claim 34 wherein said oil is administered in the
form of a food product.
37. The method of claim 34 wherein said oil is administered in an
amount from about 1 to 10 mg per kg of body weight per day to said
patient.
38. The method of claim 34 wherein total serum cholesterol is
decreased by about to 25%.
39. The method of claim 34 wherein serum LDL cholesterol is
decreased by about to 25%.
40. The method of claim 34 wherein serum HDL cholesterol is raised
by about 10 to 30%.
41. The method of claim 34 wherein the ratio of HDL to total
cholesterol in said human patient is raised from about 10 to
30%.
42. The method of claim 34 wherein the serum level of peroxides,
measured as TBARS, is decreased by about 10% to 60%.
43. The method of claim 34 wherein the serum level of tocopherol or
tocotrienol is raised by 20% to 110%.
44. A method of decreasing total blood cholesterol in a human
patient, by administering to said patient an effective amount of an
edible oil that decreases the synthesis, absorption and blood level
of cholesterol by said human patient and increases the excretion of
cholesterol from said human patient.
45. The method of claim 44 wherein said oil comprises at least one
compound that decreases cholesterol synthesis in said human
patient.
46. The method of claim 44 wherein said oil comprises at least one
compound that increases cholesterol excretion from said human
patient.
47. The method of claim 44 wherein said oil comprises a mixture of
(i) at least one tocotrienol or tocopherol; (ii) at least one free
sterol or sterol ester; (iii) at least one cycloartenol.
48. The mixture of claim 47 wherein the ratio of ingredients
(i)-(iii) of said mixture effectively decreases the absorption,
synthesis and blood levels of cholesterol by a human patient and
also increases the excretion of cholesterol from said human
patient.
49. A method of making an anti-cholesterolemic edible oil
comprising the steps of a) providing an edible oil, and b)
adjusting the content of tocopherols, tocotrienols, free sterols,
sterol esters and cycloartenols of said edible oil, such that said
oil comprises about (i) 10 to 30% of tocopherols, tocotrienols or
combinations thereof, (ii) about 2 to 20% of free sterols; (iii)
about 2 to 20% of sterol esters; (iv) about 0.1 to 1.0% of
cycloartenols, wherein all percentages are weight/weight.
50. The method of claim 49 wherein in step a) said edible oil
includes at least one saturated fat, and in step b) the content of
saturated fat of said edible oil is adjusted such that said edible
oil comprises about 7 to 19% of saturated fat.
51. The method of claim 49 wherein after step a) said edible oil is
substantially free of trans fatty acids.
Description
[0001] This application is based on U.S. Provisional Patent
Application Ser. No. 60/104,227, filed Oct. 14, 1998, the
disclosure of which is incorporated in its entirety herein by
reference.
FIELD OF THE INVENTION
[0002] The present invention relates to an edible oil that is
useful in improving blood lipid levels in a human patient, and to
methods for making and using the oil.
BACKGROUND OF THE INVENTION
[0003] More than 750,000 people in the United States die from
coronary heart disease and strokes every year. About 1.25 million
people have heart attacks every year, half of which occur without
warning. Coronary heart disease is the most frequent killer of men
and women in the United States. Despite a century of drug
development, ten times as many Americans die of heart attacks as at
the turn of the century.
[0004] According to the American Heart Association, cholesterol
levels are the major predictors of cardiovascular disease.
Cholesterol, a soft, waxy substance found among the lipids in the
blood stream, is an important part of a healthy body because it is
used to form cell membranes, some hormones and other needed
tissues. However, a high level of cholesterol in the blood
(hypercholesterolemia) is a major risk factor for coronary heart
disease, which leads to heart attack.
[0005] Cholesterol is insoluble in the blood, and must be
transported to and from the cells by a special carrier of lipids
and proteins called lipoproteins. There are several kinds of
lipoproteins, the most important of which are low-density
lipoprotein (LDL) and high-density lipoprotein (HDL).
[0006] Low-density lipoprotein is the major cholesterol carrier in
the blood. Excess LDL cholesterol circulating in the blood can
slowly build up within the walls of the arteries feeding the heart
and brain. Together with other substances it can form plaque, a
thick, hard deposit that can clog those arteries. This condition is
known as atherosclerosis. The formation of a clot (or thrombus) in
the region of this plaque can block the flow of blood to part of
the heart muscle and cause a heart attack. If a clot blocks the
flow of blood to part of the brain, the result is a stroke. A high
level of LDL cholesterol reflects an increased risk of heart
disease. Thus, LDL cholesterol is often called "bad
cholesterol."
[0007] High density lipoprotein ("HDL") carries about one-third to
one-fourth of blood cholesterol. It is believed that HDL carries
cholesterol away from the arteries and back to the liver, from
which it is ultimately passed from the body. Some experts believe
HDL removes excess cholesterol from atherosclerotic plaques and
thus slows their growth. HDL is known as "good cholesterol" because
a high level of HDL seems to protect against heart attack. The
opposite is also true: a low HDL level indicates a greater
risk.
[0008] Cholesterol comes from two sources. It is produced in the
body, mostly in the liver (about 1,000 milligrams a day), and is
also found in foods that come from animals, such as meat, poultry,
fish, seafood and dairy products. Foods from plants (fruits,
vegetables, grains, nuts and seeds) do not contain cholesterol.
[0009] Saturated fatty acids are the chief culprit in raising blood
cholesterol, which increases the risk of heart disease. But dietary
cholesterol also plays a part. The average American man consumes
about 360 milligrams of cholesterol a day; the average American
woman, between 220 and 260 milligrams.
[0010] One hundred (100) million adults have blood cholesterol
levels of 200 milligrams per deciliter (mg/dl) or higher, and
nearly 40 million Americans have levels of 240 mg/dl or above. It
is estimated that there are 53 million Americans with LDL levels
that require treatment, but that less than one-third of those in
need are receiving the necessary treatment. Furthermore, most
patients who are treated fail to attain treatment goals. The yearly
cost of treatment is estimated at more than $100 billion, yet
coronary heart disease still remains the No. 1 killer of
Americans.
[0011] Thus, the risk of having a heart attack or stroke is
strongly predicted by the amounts of low-density lipoprotein (LDL),
high-density lipoprotein (HDL), and triglycerides in the blood.
[0012] Cholesterol and triglyceride levels can be reduced through
medical intervention and/or dietary modification, such as reduction
of the dietary intake of cholesterol and saturated fats. However,
some dietary modifications have given rise to new problems. For
example, in recent years the substitution of margarine for butter
has been promoted. Butter is high in cholesterol and saturated
fats. Stick margarine, on the other hand, has a semi-solid
consistency based on their content of hydrogenated oils. The
hydrogenation process, however, forms trans fats. Clinical studies
have demonstrated that trans fats are atherogenic, causing two to
three times the cardiovascular risk of the naturally saturated fats
which give butter its stability. The health advantage of margarine
over butter is now suspect in that margarine, particularly stick
margarine, can contain 20% to 30% of trans fats. The American Heart
Association now recommends soft margarine. Such margarine, so
called trans-free margarine, which is formulated from either
completely hydrogenated palm oil or palm oil fractions, has been
introduced recently. This margarine, while free of trans fats,
contain increased levels of saturated fats, the second most
dangerous component of margarine.
[0013] Other compounds have been reported to reduce cholesterol
levels in humans. For example, plant sterols, particularly
beta-sitosterol, have been reported to have anticholesterolemic
effects, and are believed to inhibit cholesterol absorption in the
small intestine. Plant sterols are thought to displace cholesterol
in bile salt micelles. Approximately half of the dietary
cholesterol ingested is absorbed whereas less than 5% of
beta-sitosterol is absorbed. When the plant sterols displace
cholesterol of the bile salt micelles, the cholesterol is fecally
excreted.
[0014] Plant sterols exist naturally in saturated and unsaturated
forms, as free alcohols and as esters. The unsaturated forms
dominate. It is known that natural sitosterols may be converted to
sitostanols by hydrogenation, and it has been reported that stanols
are more effective per unit weight than sterols in blocking
cholesterol absorption and that stanols are not absorbed. Further,
the amount of beta-sitosterol absorbed appears to be relatively
constant even when doses administered vary by an order of
magnitude. Both sterols and stanols have been used as relative
markers of cholesterol absorption because of their unabsorbability.
However, it seems clear that while sitostanol is completely
unabsorbed, some sitosterol is.
[0015] Further, the addition of sitostanol to the diet reduces not
only cholesterol absorption but also sitosterol and vitamin
absorption. Some have characterized this as an advantage, but the
fact that sitostanols block the normal absorption of micronutrients
may be problematic.
[0016] The Lancet 1995; 345: 1529-1532, reported on the use of
beta-sitosterol (20 milligrams per day) for the treatment of benign
prostatic hyperplasia (BPH). This condition is a slow enlargement
of the fibromuscular and epithelial structures within the prostate
gland, eventually leading to obstructive urinary symptoms which are
experienced to some extent by most men over the age of 50 years.
Using sitostanols alone as an anti-cholesterolemic thus may
increase the risk of BPH.
[0017] Other compounds that have been studied in connection with
the treatment and prevention of diseases including arteriosclerosis
and high cholesterol levels include tocotrienols, which are natural
forms of vitamin E found in wheat germ, rice bran, oats and
palm.
[0018] In vitro, the concentration-dependent impact of tocotrienols
on cholesterol can be demonstrated to involve post-transcriptional
down regulation of 3-hydoxy-3-methyl-glutaryl coenzyme A reductase
(HMGCoA reductase) activity. This is the enzyme targeted by
statins, the anti-cholesterolemic drug with annual sales of eight
(8) billion dollars in the U.S. alone. Statins act directly,
blocking HMGCoA reductase. However, statins also sometimes cause
liver dysfunction.
[0019] Unfortunately, many patients taking statins or tocotrienols
respond to the decreased rate of cholesterol synthesis by a
compensatory increase in the rate at which dietary cholesterol is
absorbed from food. A recent study reports that 80% of patients
taking statins as a monotherapy failed to reach treatment goals.
With respect to statins, increasing the dosage to the levels
frequently required to overcome compensatory increase in
cholesterol absorption, produces an 11-fold increase in the
incidence of liver complications as noted above. Because of the
risk of liver complications, statins must be taken under a doctor's
supervision. Similarly, while tocotrienols have shown promise in
vitro, the results of clinical trials have been equivocal. Qureshi,
Am. J Clin. Nutr. 53: Suppl. 4: 1021S-1026S, (1991 April) reported
significant improvements in lipid parameters amongst "responders"
in a short study, but three subsequent studies of free living
patients supplemented with the same material (palm-derived
tocotrienol-rich fraction, i.e., palm-derived TRF) failed to
confirm his results. See Antila, et al, Helsinki Antioxidant
Symposium, 1991 Wahlquist, M., et al, Nutrition Research 12: Suppl.
1: S181-S201 (1992); Tomeo, A., et al., Lipids 30: 1179-1183, 19951
In response, Qureshi has suggested Qureshi, A., et al, Lipids 30
(12): 1171-1177, (1995) that d-tocopherol inhibits the
anticholesterolemic effect of tocotrienols. The results of clinical
trials with oils according to the present invention do not support
this conclusion, but rather show that synergy with other
non-saponifiable components is required to effect blood lipid
modulation.
[0020] A margarine recently introduced in Finland, Benecol, that
contains hydrogenated plant sterols extracted from pulp and paper
waste, has been found to achieve a 10-15% reduction in cholesterol
levels in patients substituting Benecol margarine for standard
margarine in their diets. This reduction corresponds to a twenty to
thirty percent decrease in cardiovascular risk. However, Benecol
suffers from the disadvantage that the plant sterol extracts
require regulatory approval in the United States and other
countries as a new food additive.
[0021] Toxic forms of oxygen have been associated with many
chronic, debilitating diseases. These include cardiovascular,
neoplastic, arthritic, age related macular degenerative and
progeria, among others. As tissue levels of these toxic forms of
oxygen rise, tissue levels of protective antioxidants, such as
antioxidants of the vitamin E family, decline. These risk factors
have been confirmed in the case of cardiovascular disease by Gey,
who showed that as blood vitamin E values decrease in a population,
the incidence of ischemic heart disease rises. To assess the blood
levels of peroxides, many researchers have measured adducts of
thiobarbituric acid (a.k.a TBARS, thiobarbituric acid reactive
substances, also called malonaldehyde modified material), or
peroxides. Holvoet, Collen and van de Werf recently documented the
relation of malonaldehydemodified LDL as a marker of acute coronary
syndromes. These scientists showed that malonaldehyde (TBARS) type
pollution in the blood indicates endothelial injury and plaque
instability, and more accurately indicates acute coronary syndromes
than other commonly used indices, such as troponin 1. In an
intervention study of patients who had had at least one stroke who
were supplemented daily with Redeem, their serum levels of TBARS
material decreased significantly from pre-study values. See: Tomeo,
A. C., el al. Antioxidant effects of tocotrienols in patients with
hyperlipidemia and carotid stenosis. Lipids 30: 1179-1183, 1995;
Watkins, T. R. et al. Hypocholesterolemic and antioxidant effects
of rice bran oil non-saponifiables in hypercholesterolemic
subjects. Environ. Nutr. Interactions, 3: (2) 1-8, 1999]. Further,
their serum vitamin E levels nearly doubled over pre-study values.
This same group of researchers of the Jordan Heart Research
Foundation had previously documented the same relation in the
laboratory rat model. See: Watkins, T. R., et al
.gamma.-tocotrienol as a hypocholesterolemic and antioxidant agent
in rats fed atherogenic diets. Lipids, 28: 1113-1118, 1993].
[0022] Accordingly, there is a need for an edible oil that is
trans-free, low in saturated fats and suitable for use in the
manufacture of margarine. Preferably, the edible oil does not
require additives that must be chemically processed (e.g.,
hydrogenated).
[0023] There is also a need for an edible oil product that is a
safe, effective alternative to known oil products and that can be
made available over-the-counter (OTC) or incorporated into staple
foods.
[0024] Furthermore, there is a need for a new intervention strategy
against cardiovascular disease, one which recognizes the difficulty
patients have in changing life-long bad eating habits and which,
unlike cardiovascular drugs, is safe enough to be taken without
direct medical supervision.
SUMMARY OF THE PREFERRED EMBODIMENTS
[0025] In accordance with one aspect of the present invention,
there is provided an edible oil that reduces the synthesis and
absorption of cholesterol by the human patient and promotes the
excretion of cholesterol from the human patient.
[0026] Preferably, the edible oil is substantially free of trans
fatty acids.
[0027] In preferred embodiments, the inventive oils are vegetable
oil or mixtures of vegetable oils. Very preferably, the inventive
oils are refined rice bran oils or mixtures of rice bran and palm
oils.
[0028] In accordance with still other aspects of the present
invention, there are provided food products that include any of the
foregoing oils.
[0029] According to additional aspects of the present invention,
there are provided methods of reducing total cholesterol and LDL
and raising HDL in a human patient comprising the step of
administering to said patient an effective amount of any of the
foregoing oils.
[0030] According to still another aspect of the present invention,
there is provided a method of making an anticholesterolemic edible
oil. The method includes the steps of providing an edible oil, and
adjusting the content of tocopherols, tocotrienols, free sterols,
steryl esters and cycloartenols of the edible oil such that the
oil, when consumed at a pre-selected dosage and in a pre-selected
dosage form, provides on a daily basis about 25 to 750 mg of
tocopherols, tocotrienols or combinations thereof, about 5 to about
500 mg of steryl esters, and about 5 to about 500 mg of
cycloartenols.
[0031] In a preferred embodiment, a crude vegetable oil ("Oil A"),
in particular a crude rice bran oil, is dewaxed and degummed, and
held under vacuum at elevated temperature. Free fatty acids are
then removed from Oil A at mild pH using an alkaline hydrous sodium
silicate and small quantities of potassium hydroxide so that the
free fatty acids are converted to soap (saponified) at conditions
which minimize the loss of esters of sterols and cycloartenols to
the soap stock. Next, a tocotrienol-rich distillate, preferably a
rice bran or palm oil deodorizer distillate, is substantially
saponified, preferably in isopropanol, and the non-saponifiable
fraction is extracted, preferably with hexane and water to yield an
extract ("Oil B"). Finally, appropriate portions of Oil A and Oil B
are mixed to produce a product having the desired concentration of
tocopherols, tocotrienols, free sterols, steryl esters and
cycloartenols.
[0032] Other objects, features and advantages of the present
invention will become apparent to those skilled in the art from the
following detailed description. It is to be understood, however,
that the detailed description and specific examples, while
indicating preferred embodiments of the present invention, are
given by way of illustration and not limitation. Many changes and
modifications within the scope of the present invention may be made
without departing from the spirit thereof, and the invention
includes all such modifications.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0033] Cholesterol levels in the human body are regulated by three
concurrent mechanisms, namely synthesis, absorption and excretion.
Most known anti-cholesterolemic compounds and compositions target
only one of these mechanisms, and thus must have a relatively large
impact on the targeted mechanism in order to function.
[0034] Edible oils according to the present invention reduce the
synthesis and absorption of cholesterol while increasing the
excretion of cholesterol. All three mechanisms are affected
simultaneously, resulting in a gentle, balanced improvement in LDL,
HDL and triglyceride levels.
[0035] Reductions and increases in cholesterol synthesis,
absorption and excretion in human patients are determined by
comparison with the same quantities measured in human patients
before and after administration of edible oils according to the
invention.
[0036] The edible oils, according to the invention, can be
characterized as "functional foods," as opposed to drugs or
nutraceuticals. Functional foods have been defined by the European
Union as "ordinary foods processed or modified in such a way that
they have scientifically documented health promoting effects and
can be marketed with a health claim." In Japan, "functional foods"
are defined as ordinary foods that are derived only from naturally
occurring ingredients and that are consumed as part of the diet and
not in supplement form (i.e., not as tablets or capsules).
[0037] Preferably, the edible oil according to the invention
includes at least one compound that reduces cholesterol synthesis
in a human patient, such as at least one tocotrienol. Specific
examples of such compounds include .alpha.-tocotrienol,
.beta.-tocotrienol, .gamma.-tocotrienol and
.delta.-tocotrienol.
[0038] The edible oil also preferably includes at least one
compound that reduces cholesterol absorption in a human patient,
for example, at least one free sterol or steryl ester. Specific
compounds useful according to the invention include ferulic and
fatty acid esters of campesterol, .beta.-sitosterol and other
sterols and stanols.
[0039] Preferably included in the inventive oil is at least one
compound that promotes cholesterol excretion in a human patient.
Such compounds include cycloartenol esters of ferulic acid
(C.sub.10H.sub.10O.sub.4), variously referred to as
3-(4-hydroxy-3-methoxyphenyl)-2-propanoic acid,
4hydroxy-3-methoxy-cinnamic acid or 3-methoxy-4-hydroxy-cinnamic
acid. Specific examples of these compounds include ferulic esters
of 24-methylene-cylcoartenol and cycloartenol.
[0040] The edible oil, according to the invention, preferably
attenuates the accumulation in and blood level of peroxidized
lipid--and other substrates, such as protein, carbohydrate and
nucleic acid--called peroxides, but also called on analysis TBARS
(thiobarbituric acid reactive substances) and malonaldehyde-like
compounds (i.e., TBA), known cardiovascular risk factors.
[0041] Very preferably, the edible oil provides at least one
compound which limits the formation and accumulation of TBARS, and
similar peroxidation adducts, in a human patient.
[0042] According to the invention, the edible oil also provides
vitamin-E like activity, whether derived from the tocotrienol or
tocopherol family, which confers antioxidant activity to tissues,
such as the blood, which can be measured as tocotrienol or
tocopherol.
[0043] And, very preferably, at least one compound in the oil is
derived from the tocotrienol or tocopherol family (as described in
detail herein), which results in elevated serum tocopherol or
tocotrienol levels in the blood of a human patient at risk of
cardiovascular disease.
[0044] The tocopherols, tocotrienols, sterols, steryl esters and
cycloartenols employed according to the invention are preferably
derived from natural sources, but can also be synthetically
produced, if desired. In particular, one or more of the ingredients
can be synthetic or can be derived from a source other than the
vegetable oil base.
[0045] Very preferably, the inventive oil is substantially free of
trans fatty acids. "Substantially free" as used herein means less
than about 2% (weight/weight). Optimally, the inventive oil
includes no trans fatty acids.
[0046] It has been discovered in clinical trials that the optimal
ratio of (i) tocotrienols and/or tocopherols to (ii) free sterols
and/or steryl esters to (iii) cycloartenols ranges from about
1:0.5:0.05 to 1:5:0.5, and very preferably is about 1:1:0.05. Thus,
in a preferred embodiment, the ratio of the foregoing ingredients
in the inventive oil falls within this preferred range, and
optimally is about 1:1:0.05.
[0047] The amount of tocopherols and tocotrienols administered to a
human patient preferably ranges from about 50 to 500 mg per day, or
alternatively, about 10 to 200 mg/dosage unit. The amount of
sterols and steryl esters likewise preferably ranges from about 50
to 500 mg/day. The amount of cycloartenols preferably ranges from
about 2.5 to 25 mg per day. In a preferred embodiment, a human
patient is administered about 400 milligrams of tocopherols and
tocotrienols, 400 milligrams of sterols and steryl esters and 20
milligrams of cycloartenols. This corresponds to the preferred
ratio of ingredients of about 1:1:0.05. The percentages of the
various ingredients in the inventive oil can vary within a wide
range, so long as the proportions of the ingredients are within the
stated ranges and the patient consumes a total amount of each
ingredient within the stated ranges each day.
[0048] In a preferred embodiment, the edible oil according to the
invention is a vegetable oil, in particular a refined rice bran oil
or a mixture of refined rice bran oil and palm oil.
[0049] Very preferably, the edible oil is a refined rice bran oil.
Crude rice bran oil contains the highest percentage of
non-saponifiables of any commercial vegetable oil. Total
non-saponifiables often exceed 4% (by weight), approximately four
times more than the oils currently used in margarine
manufacture.
[0050] Crude rice bran oil is preferably refined for use according
to the invention. When the oilbearing bran is separated from rice,
a particularly active lipase enzyme is activated which causes a
very rapid increase in free fatty acids. Even when the rice is
stabilized by heat or chemicals shortly after milling, free fatty
acid (FFA) levels of 5 to 10% are common; industrial rice bran oils
may have FFA levels as high as 30%. There is furthermore, a
seasonal variation in FFA levels. Wax levels are also high and
generally related to the temperature at which the bran is extracted
with solvent. Rice bran oil thus is among the most challenging of
oils to refine. Industrially, rice bran oil is processed by
chemical refining.
[0051] Palm oil, another useful source of tocotrienols, also has
high FFA levels because palm fruit releases a lipase enzyme when
bruised. FFA levels of palm oil range from 2% to 5%. Most
commercial production of palm oil uses physical refining processes.
However, such refining methods produce low-grade distillates of low
tocopherol/tocotrienol concentration, typically 3,000-5,000 ppm.
Deodorizer distillates obtained from chemical refineries are of
higher concentration, typically 1 to 3% tocopherol/tocotrienol.
However, during the chemical refining process, the ferulic and
fatty acid esters of sterols and cycloartenols are ionized and lost
to the soap stock. Free sterols, and triterpene alcohols
(cycloartenols) are soluble to various extents in both polar and
non-polar solvents, whereas non-polar solvents are selective with
respect to their esters of fatty acids and ferulic acids. These
molecules are structurally similar to cholesterol. In the ester
form they more readily displace cholesterol from the micelles in
the digestive tract, but are themselves not absorbed, or if
absorbed, quickly excreted.
[0052] Thus, while a tocotrienol-rich fraction can be recovered
from chemically refined rice deodorizer distillate, it is
substantially depleted of these useful esters.
[0053] Known processes for production of tocopherol and
tocotrienol-rich fractions from deodorizer distillates include
processes such as ion exchange, saponification and extraction from
hard soap, methyl esterification, esterification of free fatty
acids and molecular distillation, and desterolization. In each of
these processes, the method of separation of tocotrienol-rich
concentrates purposely or incidentally removes the natural steryl
esters of ferulates and cycloartenols.
[0054] As crude rice bran oil ages, it has been observed that the
free sterols become esterified with free fatty acids. It has been
discovered that this process can be accelerated by aging the oil at
elevated temperatures under vacuum, under mild conditions such that
the free sterols and free cycloartenols are esterified and the
tocopherols are not. Rogers, et al, J. Am. Oil Chem. Assoc., 70:
No. 3, 1993, analyzed five commercially available rice bran oils
from different manufacturers. The content of ferulic esters of
cycloartenols and plant sterols (quantitated as oryzanols) ranged
between 115 ppm and 787 ppm, with an average of 400 ppm. In the
same oils, tocotrienol content ranged between 72 ppm and 1157 ppm,
with an average of 500 ppm. More than 95% of the oryzanols and 60%
of the tocotrienols are lost in conventional refining processes. To
obtain effective amounts of the anticholesterolemic active
principles, a patient could be required to consume more than a
kilogram of oil per day.
[0055] The invention thus meets the need for a new process in which
the optimal proportions of tocotrienols, steryl esters and
cycloartenols are retained in the product and the sterols present
are substantially in the form of steryl esters whose increased
solubility in lipids underlies the efficacy of the compound in
decreasing the absorption of dietary cholesterol.
[0056] In general, the inventive method begins with crude rice bran
oil ("Oil A"), which is dewaxed and degummed, and free sterols and
triterpene alcohols esterified with free fatty acids. The remaining
free fatty acids are then removed under conditions that preserve
the esterified state of the sterols and cycloartenols, by
distillation or at mild pH using an alkaline hydrous sodium
silicate and small quantities of potassium hydroxide so that the
free fatty acids are converted to soap (saponified) at conditions
which minimize the loss of esters of sterols and cycloartenols to
the soap stock. Next, a tocotrienol-rich deodorizer distillate,
preferably of rice bran or palm oil, is substantially saponified in
isopropanol and the non-saponifiable fraction is extracted with
hexane and water to produce and extract ("Oil B"). Finally,
appropriate portions of Oil A and Oil B are mixed to form a product
having the required concentration of tocopherols, tocotrienols,
free sterols, steryl esters and cycloartenols.
[0057] In contrast to products such as Benecol, which include
synthetic ingredients requiring regulatory approval, preferred
embodiments of the edible oils according to the present invention,
such as refined rice bran oil, raise no regulatory issues since
they naturally contain suitable steryl and stanyl esters, which,
unlike Benecol, do not require hydrogenation and chemical
processing with attendant risks of trans fatty acid formation.
[0058] The edible oils according to the invention can be
incorporated into a variety of food products, including, without
limitation, butter, margarine, ice cream and mayonnaise--chocolate
products; liquid such as soybean milk and rice milk--and
water-based drinks such as wines and mineral waters. The inventive
oils are also suitable for encapsulation in gelatin shells to form
soft gels. Regardless of the particular form in which the inventive
oil is prepared, the daily dosage of the various ingredients to a
human patient should fall within the ranges set forth above.
Depending on the concentration of the inventive oil in the given
form, the total amount of the food product per serving, or
encapsulated oil, etc., will also vary. Highly concentrated forms,
such as soft gels, will be administered in lower total amounts than
diluted forms, such as drinks.
[0059] The invention is further illustrated by the following
non-limiting examples.
[0060] Example 1 discloses a method for the preparation of an
embodiment of the inventive oil. Examples 2, 3, 4 and 5 compare the
results of human studies in which patients received the inventive
oil or other preparations containing the elements of the inventive
oils individually or in proportions which differ substantially from
those of the invention. Example 2 compares the administration of
palm-derived tocotrienols with the inventive oil. Example 3
compares rice tocotrienols processed by another method with the
inventive oil. Example 4 compares a conventionally processed rice
oil with the inventive oil. Example 5 compares the performance of
the inventive oil against two commercial margarines incorporating
high levels of sterols and steryl esters.
[0061] In each case, it is demonstrated that the desired changes in
blood lipid values are achieved only when the proportions of the
active components are according to the invention. It is shown that
the inventive oil is markedly superior, decreasing LDL levels and
uniquely superior in elevating HDL and diminishing triglyceride
values.
EXAMPLE 1
[0062] One hundred (100) grams of rice oil ("Oil A") are analyzed,
dewaxed and degummed, and the acid value of the oil is determined
by AOAC methods. The oil contains 5% sterols, stanols, and
cycloartenols as:
1 Campesterol 15% Sitosterol 10 Campestanol 1.4 Stigmasterol 1.5
Sitostanol 1.5 Gycloartenol 30 24-methylene- 40 cycloartenol
[0063] and, further, contains 1120 ppm of tocopherols and
tocotrienols, 58% as gamma-tocotrienol. Oil A is held overnight at
moderate vacuum at 125.degree. C., so that water generated during
esterfication is removed and the reaction driven to the right,
accelerating the natural aging process in which fatty acid esters
of sterols and cycloartenols are formed. Oil A is then cooled to
50.degree. C.
[0064] A mild, caustic agent is prepared by combining potassium
hydroxide and alkaline hydrous sodium silicate (Britesorb.RTM. NC,
commercially available from PQ Corporation, Valley Forge, Pa.) in a
slurry, in the ratio of 1 part potassium hydroxide to 4 parts
Britesorb.RTM. to 6 parts of water. An amount of slurry equal to 5%
stoichiometric excess of the acid value previously measured is
added to the cooled Oil A above, and the mixture is stirred at
60.degree. C. for one hour, after which the temperature is
increased to 80.degree. C. and the mixture filtered.
[0065] The refined oil is washed and dried, leaving a neutral oil
rich in sterol esters and cycloartenol esters of ferulic acids, but
substantially free of free fatty acids and free sterols.
[0066] Next, a deodorizer distillate from chemical refining of rice
bran oil is obtained and analyzed. The distillate is found to
contain the following ingredients:
2 2.0% tocopherols and tocotrienols, including: .alpha.-tocopherol
0.5% .gamma.-tocopherol 0.4% .alpha.-tocotrienol 0.1%
.gamma.-tocotrienol 1.0% 8.0% sterols, total 2.0% steryl esters
75.2% glycerides, total, including: free fatty acids 43.5%
monoglycerides 6.7% diglycerides 8.6% triglycerides 16.4% The
distribution of fatty acids is as follows: C12:0 0.1% (by weight)
C14:0 1.0 C16:0 27.5 C16:1 0.3 C18:0 2.0 C18:1 39.0 C18:2 27.0
C18.3 0.8 C20:0 0.8 C20:1 0.6 C22:0 0.2 C24:0 0.4
[0067] Next, 50 grams of the distillate is mixed with 5 volumes of
isopropanol and the saponification calculated from the analysis
above using the AOAC method. Then an amount of 80% potassium
hydroxide at 150% of the calculated stoichiometric weight required
to saponify all of the glyceride components is determined, and is
added slowly to the reaction mixture. The reaction mixture is held
in a water bath for 30 minutes at 60.degree. C. and allowed to
cool, then neutralized and extracted with 10 volumes of hexane and
20 volumes of water over night. The hexane phase is separated,
washed and dried, and the semi-solid resultant phase ("Oil B") is
combined with the neutral oil obtained above. The resultant refined
oil is enriched in tocopherols and tocotrienols, and esters of
cycloartenols and sterols, but depleted of free fatty acids, mono-,
di- and triglycerides and free sterols.
[0068] Optionally, the semi-solid resultant phase above can be
de-sterolized by precipitating free sterols from methanol in
4.degree. C., and further concentrated by distillation, prior to
blending.
[0069] The yield of tocopherols and tocotrienols ranges between 45%
and 75%, depending upon the degree of saponification of the
reaction mixture. The ratio of tocotrienols and tocopherols to
cycloartenols and ferulic and sterol esters can also be adjusted by
the degree of saponification.
[0070] The ratio of Oil A and Oil B combined can be varied to
produce a re-proportioned oil ranging in concentration of
tocopherols and tocotrienols between about 0.5% and 25%
(weight/weight, based on the total weight of the oil). At higher
concentrations, the oil is suitable for encapsulation into soft
gels as a nutraceutical or therapeutic. At lower concentrations,
the oil can be incorporated directly into food products, such as
margarine or mayonnaise. In each case, the desired concentration is
that sufficient to provide between 50-500 mg/day (or,
alternatively, about 20-200 mg/serving) of tocotrienols/tocopherols
to a patient consuming the product.
EXAMPLE 2
[0071] A test group was administered a palm oil-derived
tocotrienol-rich fraction (TRF), processed in a manner which
depletes sterols, steryl esters and cycloartenols, in an amount of
160-240 mg three times per day for one year. Tomeo, A., et al
Lipids 30: 1179-1183, 1995. The dosage was increased to 240 mg of
tocotrienols three times per day for two additional years.
Kooyenga, D., et al. Asia Pacific J Clin. Nutr. 6: 72-75, 1996. No
change in total cholesterol, LDL or HDL cholesterol, or
triglyceride levels was observed for two years. The test group was
then administered 2.4 grams per day of an oil according to the
invention containing 200 mg of tocotrienols three times per day for
one year. The blood lipids improved: a 20% decrease in LDL
cholesterol, a 20% increase in HDL cholesterol and a 23% decrease
in triglycerides was observed. See Table 1.
3TABLE 1 Changes in blood lipids during supplementation with
palm-derived tocotrienol or the invention, rice bran tocotrienols
with non-saponifiables. Data in mg/dl. N = 50 subjects. Baseline
(start) Year 1 Year 3 Year 4 Palm tocotrienol The Lipid period
invention Cholesterol, total 239 239 239 206* LDL cholesterol 165
165 165 132* HDL cholesterol 40 40 40 48* Triglycerides 211 211 211
162* *Data of years 3 and 4 differ significantly, p < 0.01.
EXAMPLE 3
[0072] Ten members of the study of Example 2 were switched to
NuTriene.RTM., a tocotrienol-rich concentrate from rice oil
manufactured by Eastman Chemical Company, processed in a manner
that depletes the oil of steryl esters and cycloartenols. The
lowering of LDL cholesterol observed in Example 2 was maintained,
but 25% of the increase in HDL cholesterol levels and 50% of the
decrease in triglycerides was lost. See Table 2.
4TABLE 2 Changes in blood lipids in subjects supplemented with the
invention, rice bran tocotrienols and non-saponifiables, or
NuTriene .RTM. , between years 4 and 5. Data as mg/dl. N = 10.
Lipid Year 4 (end) Year 5 (end) Cholesterol, total 206 205 LDL
cholesterol 132 127 HDL cholesterol 48 46 Triglycerides 162 187
EXAMPLE 4
[0073] Patients were administered a margarine prepared from a
physically refined rice oil rich in sterols, steryl esters and
cycloartenois (1.5 grams total per day) but poor in tocotrienols
(2.5 mg per day). No change in blood lipid values was observed.
Weststrate, J. A., et al European J. Clin. Nutr. 52: 334-343, 1998.
See Table 3.
5TABLE 3 Percent change in blood lipid components in patients
administered sterol and stanol esters and tocotrienols from a
physically refined rice bran oil compared with those obtained with
the inventive oil. Lipid Rice bran oil* The invention**
Cholesterol, total -1.1 -14 LDL cholesterol -1.5 -20 HDL
cholesterol -1.3 +20 LDL/HDL cholesterol -0.3 +28 *1.5 g/d in 30 g
oil providing 2.5 mg/d tocotrienols. **As described above.
EXAMPLE 5
[0074] The patients of Example 4 were switched to margarine
preparations containing either stanol esters (Benecol) or steryl
esters. Both groups received margarine containing approximately 3
grams per day of the stated esters. Decreases of LDL cholesterol
concentrations between 8% and 13% were observed, but no significant
changes in HDL cholesterol or triglycerides were found. This study
confirms the findings of other studies of sitostanol esters but
reported significant depletion of plasma antioxidants. Miettinen,
T. A. et al New Engl. J Med 333: 1308-1312, 1995; Weststrate, J.
A., et al. Eur. J Clin. Nuir. 52: 334-343, 1998. See Table 4.
6TABLE 4 Percentage change in blood lipids in patients administered
sterol and stanol esters compared with those obtained with the
inventive oil. Lipid Steryl esters* Stanol esters** The inventive
oil*** Cholesterol, total -8.3 -7.3 -14 LDL cholesterol -13 -13 -20
HDL cholesterol n.s. n.s. +20 Trigiycerides n.s. n.s. -23 LDL/HDL
chol. -14 -12 -28 *Steryl esters: 3 g/d in 30 g oil; 0 mg/d
tocotrienols. **Stanol esters: 3 g/d in 30 g oil; 0 mg/d
tocotrienols. *** As described above.
Conclusion
[0075] The foregoing data demonstrate that the control of the
amounts of tocotrienols/tocopherols, steryl esters, free sterols,
and cycloartenols according to the invention modulate blood lipid
values in an optimally therapeutic manner. The results in Example 2
show that the administration of tocotrienols alone does not affect
blood lipid values. Example 2 further demonstrates that
administration of an edible oil according to the invention lowers
total cholesterol, LDL cholesterol and triglycerides, while
increasing HDL cholesterol. Example 3 demonstrates that a
rice-derived tocotrienol concentrate depleted in steryl esters and
cycloartenols is ineffective in increasing HDL levels or decreasing
triglycerides. Example 4 shows that a physically refined rice oil
poor in tocotrienols is ineffective, and Example 5 shows that
administration of sterols and steryl esters without tocotrienols
does not result in changes in HDL or triglyceride values.
[0076] The decreases in serum total cholesterol and LDL cholesterol
achieved by patients administered the oil according to the
invention are comparable to the best results achieve by drug
therapy. The increase in HDL cholesterol levels of 20% and the
decrease in triglycerides of 23% are particularly significant.
Watkins, T. et al.
[0077] Environmental Nutr. Interactions 3:8-18, 1999. The total
cholesterol (TC)/HDL cholesterol ratio has been found to be a
superior measure of risk of coronary heart disease compared with
either total cholesterol or LDL cholesterol levels alone. In the
Framingham study, the eight-year likelihood ratios for coronary
heart disease increased 10 times in men with the lowest TC/HDL
ratios compared with men with the highest ratios. Kinosian, B. et
al L J Invest Med 43.-443-450, 1995. On average, the TC/HDL ratio
of patients administered the oil according to the invention
decreased by 28% from 5.97 to 4.29. The potential impact of the
inventive oil to improve the public health is self-evident. A
fifty-year-old man who achieves the 28% reduction in TC/HDL ratios
reported in the clinical trial with the inventive oil is twice as
likely to live beyond the age of 75 years.
[0078] The foregoing examples are for illustrative purposes only,
and do not in any way limit the scope of applicants' invention
which is identified by the claims appended below.
* * * * *