Note: Descriptions are shown in the official language in which they were submitted.
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
COMPOSITIONS, KITS, AND METHODS FOR CARDIOVASCULAR HEALTH
REFERENCE TO PRIORITY APPLICATION
The present invention claims priority to U.S. Provisional Application Serial
No.
60/178,778, filed January 28, 2000.
FIELD OF THE INVENTION
The present invention relates to compositions, kits, and methods which are
useful for providing various general health benefits including, but not
limited to cardiac
benefits, including lowering cholesterol in the consumer, treating,
preventing, and / or
inhibiting heart disease (e.g., atherosclerosis, restenosis, thrombosis) and
treating
conditions such as hypercholesterolemia, hypertension, poor circulation, and
complications associated with diabetes.
BACKGROUND OF THE INVENTION
Cardiovascular conditions, including heart disease, hypercholesterolemia,
hypertension, poor circulation, and complications associated with diabetes,
are serious
medical conditions which are leading causes of mortality in humans. Various
regimens
have been suggested for prevention and treatment of these conditions,
including
pharmaceutical, dietary, and exercise regimens. Notwithstanding, they remain
among
the most prevalent and serious of all medical conditions.
L-arginine is a natural amino acid which has been identified to provide
certain
general health benefits including, for example, cardiovascular benefits, such
as
lowering cholesterol in the consumer, and treating, preventing, and / or
inhibiting heart
disease and poor circulation. See e.g., Moskowit~, U.S. Patent No. 5,385,940,
assigned
to The General Hospital Corp., issued January 31, 1995; Sonaka et al., EP
0,546,796,
assigned to Ajinomoto Co., published June 16, 1993; Cotter et al., U.S. Patent
No.
4,920,098, assigned to Baxter International Inc., issued April 24, 1990;
Dudrick, U.S.
Patent No. 5,032,608, issued July 16, 1991; Levere et al., U.S. Patent No.
5,217,997,
issued June 8, 1993; Cooke et al., U.S. Patent No. 5,428,070, assigned to
Stanford
University, issued June 27, 1995; Chibata et al., U.S. Patent No. 4,420,432,
assigned to
Tanabe Seiyaky Co., issued December 13, 1983; Varma et al., U.S. Patent No.
1
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
5,364,884, assigned to Baylor College of Medicine, issued November 15, 1994;
and
Barbul, U.S. Patent No. 5,157,022, issued October20, 1992.
The utility of L-arginine, particularly to advance cardiovascular health, is
therefore well known in the art. However, as for any beneficial regimen,
compliance
must be assured in order to realize the various benefits thereof.
Unfortunately, L-
arginine and its close derivatives (including salts, polypeptides, and pro-
forms) have a
strong, bitter, and fishy flavor, making L-arginine generally unacceptable for
use. This
results in decreased compliance of a regimen involving L-arginine, and the
requisite
cardiovascular benefits are therefore not realized. Accordingly, to enhance
compliance,
it would be desirable to provide L-arginine in a form which diminishes and /
or removes
the unacceptable flavor associated with L-arginine.
Unfortunately, flavor improvement is typically associated with a decrease in
the
general health benefits of the component which is desired to be delivered.
Additionally,
because delivery of relatively large amounts of L-arginine is desirable (e.g.,
about 3
grams to about 10 grams of L-arginine per dose), it becomes increasingly more
difficult
to mask the strong, bitter, and fishy flavor. Such difficulties manifest
themselves in the
marketplace, where it is understood that current products containing L-
arginine are not
acceptable to the consumer due to unacceptable flavor.
The present inventors have surprisingly discovered that the unacceptable
flavor
of L-arginine is significantly improved through combination with a second
component,
which is described herein as a sterol, stanol, ester thereof, or a polyol
fatty acid
polyester. Interestingly, and quite unexpectedly, this second component
diminishes
and / or removes the unacceptable flavor associated with the L-arginine.
Accordingly,
such combination is acceptable to consumers which, more importantly,
translates into
improved regimen compliance and enhanced cardiovascular, and other health,
benefits.
Additionally, the second component does not decrease the cardiovascular health
benefits -of the resulting composition, but rather enhances such benefits. For
example,
sterols, stanols, and their esters have been utilized in food compositions to
decrease
cholesterol. Similarly, polyol fatty acid polyesters (e.g., sucrose
polyesters) add no fat
to the composition and may reduce cholesterol, but maintain flavor properties
of
traditional fat products and, in this case, improve the overall flavor of the
composition by
diminishing and / or removing the unacceptable flavor of the L-arginine.
2
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
The foregoing findings are unexpected relative to the known literature.
Accordingly, the present inventors have discovered compositions which provide
general
health benefits, including cardiovascular benefits. Relative to known
products,
compliance is improved and / or ensured through use of such compositions
because
the flavor is acceptable to the consumer. The compositions are easily provided
as a
pharmaceutical or food product (preferably, a food product) and may be
delivered in kit
form, wherein the kit has the further advantage of disseminating information
to the
consumer regarding various health benefits and dose regimens of the
composition.
SUMMARY OF THE INVENTION
The present invention is directed to compositions comprising:
(a) a first component selected from the group consisting of L-arginine,
polypeptides thereof, acceptable salts thereof, pro-forms thereof, and
mixtures thereof; and
(b) a second component selected from the group consisting of sterols, stanols,
esters thereof, polyol fatty acid polyesters, and mixtures thereof.
The present invention is further directed to kits comprising these
compositions
as well as methods of using the compositions. The compositions, kits, and
methods
herein are useful for providing general health benefits to the consumer,
particularly
cardiovascular benefits, anti-menopausal benefits and l or treating sexual
dysfunction
(particularly, erectile dysfunction). Most particularly, the compositions,
kits, and
methods herein are useful for providing cardiovascular benefits, including
lowering
cholesterol in the consumer, treating, preventing, and / or inhibiting heart
disease (e.g.,
atherosclerosis, restenosis, thrombosis) and, for example, treating other
conditions
such as hypercholesterolemia, hypertension, poor circulation, and
complications
associated with diabetes.
DETAILED DESCRIPTION OF THE INVENTION
The present invention is directed to compositions which are useful for
providing
general health benefits to the consumer, particularly cardiovascular benefits,
anti-
menopausal benefits and l or treating sexual dysfunction (particularly,
erectile
dysfunction). The invention herein is further directed to kits comprising the
3
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
compositions and methods of using the compositions to provide the foregoing
general
health benefits.
Publications, patents, and patent applications are referred to throughout this
disclosure. All references cited herein are hereby incorporated by reference.
All percentages and ratios are calculated by weight unless otherwise
indicated.
All percentages and ratios are calculated based on the total composition
unless
otherwise indicated.
All component or composition levels are in reference to the active level of
that
component or composition, and are exclusive of impurities, for example,
residual
solvents or by-products, which may be present in commercially available
sources.
Referred to herein are trade names for components including, but not limited
to,
certain carbohydrates, flavors, and other components. The inventors herein do
not
intend to be limited by materials under a certain trade name. Equivalent
materials (e.g.,
those obtained from a different source under a different name or catalog
(reference)
number) to those referenced by trade name may be substituted and utilized in
the
compositions, kits, and methods herein.
In the description of the invention various embodiments and/or individual
features are disclosed. As will be apparent to the ordinarily skilled
practitioner, all
combinations of such embodiments and features are possible and can result in
preferred executions of the present invention.
The compositions, methods, and kits herein may comprise, consist essentially
of, or consist of any of the elements as described herein.
Compositions of the Present Invention
The present invention is directed to compositions which are useful for
providing
general health benefits to the consumer, particularly cardiovascular benefits,
anti-
menopausal benefits and / or treating sexual dysfunction (particularly,
erectile
dysfunction). The invention herein is further directed to kits comprising the
compositions and methods of using the compositions to provide the foregoing
general
health benefits. Most particularly, the compositions, kits, and methods herein
are useful
for providing cardiovascular benefits, including lowering cholesterol in the
consumer,
treating, preventing, and / or inhibiting heart disease (e.g.,
atherosclerosis, restenosis,
4
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
thrombosis) and, for example, treating other conditions such as
hypercholesterolemia,
hypertension, poor circulation, and complications associated with diabetes.
The compositions herein comprise:
(a) a first component selected from the group consisting of L-arginine,
polypeptides thereof, acceptable salts thereof, pro-forms thereof, and
mixtures thereof; and
(b) a second component selected from the group consisting of sterols, stanols,
esters thereof, polyol fatty acid polyesters, and mixtures thereof.
The present inventors have discovered that such compositions are particularly
useful for delivering cardiovascular benefits through a synergistic
combination of the L-
arginine (including polypeptides, acceptable salts, and pro-forms thereof) and
the
sterols, stanols, esters thereof, or a polyol fatty acid polyester. As a
further beneficial
aspect of these compositions, the present inventors have surprisingly
discovered that
the undesirable flavor of L-arginine is significantly diminished or removed
through
combination with the second component. This surprising and unexpected results
allows
for enhanced delivery and compliance associated with ingestion of L-arginine
for
various health benefits, while additionally providing the health benefits
known to be
associated with the second component.
As used herein, the first component is selected from L-arginine, polypeptides
thereof, acceptable salts thereof, pro-forms thereof, and mixtures thereof.
Preferably,
the first composition is selected from L-arginine and salts thereof. As
further used
herein, the second component is selected from sterols, stanols, esters
thereof, polyol
fatty acid polyesters, and mixtures thereof. The terms "first component" and
"second
component" are utilized herein strictly for convenience of reference and in no
manner
are these terms intended to limit order of addition to the composition,
importance of the
various components, and any other limiting factors.
First Component
The first component of the present compositions is selected from the group
consisting of L-arginine, polypeptides thereof, salts thereof, pro-forms
thereof, and
mixtures thereof. L-arginine is a natural amino acid which has been identified
to
provide certain general health benefits including, for example, cardiovascular
benefits,
including lowering cholesterol in the consumer, and treating, preventing, and
l or
inhibiting heart disease (e.g., atherosclerosis, restenosis, hypertension,
poor circulation,
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
and / or complications associated with diabetes. See e.c~., Moskowitz, U.S.
Patent No.
5,385,940, assigned to The General Hospital Corp., issued January 31, 1995;
Sonaka
et al., EP 0,546,796, assigned to Ajinomoto Co., published June 16, 1993;
Cotter et al.,
U.S. Patent No. 4,920,098, assigned to Baxter International Inc., issued April
24, 1990;
Dudrick, U.S. Patent No. 5,032,608, issued July 16, 1991; Levere et al., U.S.
Patent
No. 5,217,997, issued June 8, 1993; Cooke et al., U.S. Patent No. 5,428,070,
assigned
to Stanford University, issued June 27, 1995; Chibata et al., U.S. Patent No.
4,420,432,
assigned to Tanabe Seiyaky Co., issued December 13, 1983; Varma et al., U.S.
Patent
No. 5,364,884, assigned to Baylor College of Medicine, issued November 15,
1994; and
Barbul, U.S. Patent No. 5,157,022, issued October20, 1992.
Wherein L-arginine, a polypeptide thereof, salt thereof, or mixture thereof is
utilized in the compositions, typically from about 0.0001 % to about 25%, by
weight of
the composition, is utilized in such composition. More preferably from about
0.1 % to
about 20%, even more preferably from about 1 % to about 15%, and most
preferably
from about 3% to about 10%, by weight of the composition, is utilized in such
composition. Additionally, as a daily dose is frequently important for
maintenance of
the general health benefits provided by the L-arginine, typically from about
0.05 grams
to about 50 grams of the L-arginine, polypeptide thereof, salt thereof, or
mixture thereof
is administered daily in such composition. More preferably, from about 0.01
grams to
about 20 grams, even more preferably from about 0.1 gram to about 10 grams,
and
most preferably from about 0.5 grams to about 6 grams of the L-arginine,
polypeptide
thereof, salt thereof, or mixture thereof is administered daily in such
composition.
The L-arginine utilized herein as the first component may be used in its free
form
or may be utilized as a salt, a polypeptide, and / or a pro-form. Salts of L-
arginine are
particularly preferred herein as they typically provide enhanced palatability
relative to
the free form of L-arginine. The salt used herein should be an acceptable
salt; i.e., a
salt useful in pharmaceutical and / or food compositions, preferably food
compositions.
Many suitable salts of L-arginine are commonly known to one of ordinary skill
in the art.
For example, Greenbera et al., U.S. Patent No. 5,780,039, assigned to Novartis
Nutrition, issued July 14, 1998 discloses palatable forms of L-arginine as
acceptable
salts. Such salts include those of food grade acids such as phosphoric,
citric, adipic,
tartaric, acetic, fumaric, malic, and lactic acid. Thus, as non-limiting
examples
phosphate, citrate, acetate, malate, tartrate, fumarate, adipate, and lactate
salts of L-
6
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
arginine may be utilized as the first component herein. Additionally, the
hydrochloride
salt of L-arginine may be similarly utilized. The acetate and hydrochloride
salts of L-
arginine are particularly preferred.
Polypeptides of L-arginine are also well-known in the art. Preferred
polypeptides for use herein include those which are readily hydrolyzed in vivo
to provide
free L-arginine. Dipeptides and tripeptides of L-arginine are particularly
preferred. Pro-
forms of L-arginine may also be utilized herein. Pro-forms (also commonly
referred to
as pro-drugs) are those forms which, upon hydrolysis in vivo, provide the free
L-
arginine. Non-limiting, but preferred, examples of such pro-forms include the
esters
and amides of L-arginine, for example, L-arginine methyl, ethyl, propyl, or
butyl ester,
preferably methyl ester. Amides of the ~-nitrogen of L-arginine are also
particularly
useful as pro-forms herein.
Second Component
The second component of the present compositions is selected from sterols,
stanols, esters thereof, polyol fatty acid polyesters, and mixtures thereof.
By "esters
thereof" it is meant that sterol esters and stanol esters are included within
the definition
of the second component.
Sterols, Stanols, and Esters Thereof
The second component may be a sterol, stanol, ester thereof, or mixtures
thereof. Such sterols, stanols, and esters have recently been identified as
useful for
certain cardiovascular benefits, including lowering cholesterol. The present
inventors
have surprisingly discovered that combination of such sterols, stanols, and /
or esters
with the first component herein provides several unexpected benefits. For
example, the
sterols, stanols, and / or esters (particularly wherein esters are included)
encapsulate
the first component to provide sustained delivery of the first component.
Additionally,
the combination also diminishes and l or removes the adverse flavor typically
associated with the first component. As a further advantage, the sterols,
stanols, and /
or esters interact synergistically with the first component to provide the
foregoing
cardiovascular benefits.
In a particularly preferred embodiment herein, the present compositions
comprise a mixture of at least one sterol or stanol and at least one sterol
ester or stanol
ester. In a more preferred embodiment of the present invention, the
composition
comprises a mixture selected from: a) a mixture of one or more sterols and one
or more
7
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
sterol esters; and b) a mixture of one or more stanols and one or more stanol
esters. In
the most preferred embodiment herein, the composition comprises a mixture
selected
from: a) a mixture of one or more sterols and one or more sterol fatty acid
esters; and b)
a mixture of one or more stanols and one or more stanol fatty acid esters.
Without
intending to be limited by theory, the present inventors have discovered that
the
foregoing mixtures encapsulate (coat) the first component which further
disguises the
adverse flavor of the first component when administered orally. Additionally,
sustained
delivery of the first component is accomplished through such encapsulation,
providing
enhanced and prolonged bioavailability of the first component to provide the
requisite
health benefits.
Wherein a mixture of at least one sterol or stanol and at least one sterol
ester or
stanol ester is utilized, the ratio of the sterolslstanols relative to the
sterol/stanol esters
can be important. Preferably, the ratio of sterols and stanols to the esters
is from about
99:1 to about 1:99. More preferably, the ratio of sterols and stanols to the
esters is
from about 75:25 to about 25:75. Most preferably, the ratio of sterols and
stanols to
the esters is from about 60:40 to about 40:60. In all of the foregoing, such
ratios are
calculated by weight of the sterols, stanols, and esters.
Wherein a sterol, stanol, ester thereof, or mixture thereof is utilized in the
compositions, typically from about 0.0001 % to about 25%, by weight of the
composition, is utilized in such composition. More preferably from about 0.1 %
to about
20%, even more preferably from about 1 % to about 15%, and most preferably
from
about 3% to about 10%, by weight of the composition, is utilized in such
composition.
Additionally, as a daily dose is frequently important for maintenance of the
general
health benefits provided by the sterol, stanol, or ester, typically from about
0.01 grams
to about 50 grams of the sterol, stanol, ester, or mixture thereof is
administered daily.
More preferably, from about 0.05 grams to about 20 grams, even more preferably
from
about 0.1 gram to about 6 grams, and most preferably from about 0.2 grams to
about 4
grams of the sterol, stanol, ester, or mixture thereof is administered daily.
Sterols, stanols, and esters thereof (particularly fatty acid esters thereof),
which
are useful as the second component herein, are commonly known in the art. As
non-
limiting examples, such second components are described in Stern, U.S. Patent
No.
3,004,043, assigned to Eastman Kodak Co., issued October 10, 1961; Wruble et
al.,
U.S. Patent No. 3,085,939, issued April 1, 1963; Erickson, U.S. Patent No.
3,751,569,
8
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
assigned to The Procter & Gamble Co., issued August 7, 1973; Jandacek, U.S.
Patent
No. 3,865,939, assigned to The Procter & Gamble Co., issued February 11, 1975;
Ong,
U.S. Patent No. 4,195,084, assigned to Eli Lilly and Co., issued March 25,
1980;
Malinow, U.S. Patent No. 4,461,762, assigned to Medical Research Foundation,
issued
July 24, 1984; Arichi et al., U.S. Patent No. 4,524,067, assigned to Osaka
Chemical
Lab. Co., issued June 18, 1985; Malinow, U.S. Patent No. 4,602,003, assigned
to
Medical Research Foundation, issued July 22, 1986; Cassal, U.S. Patent No.
4,680,290, assigned to Hoffman-La Roche Inc., issued July 14, 1987; Ambrus et
al.,
U.S. Patent No. 5,112,815, issued May 12, 1992; Straub, U.S. Patent No.
5,244,887,
issued September 14, 1993; Eugster et al., U.S. Patent No. 5,270,041, assigned
to
Marigen S.A., issued December 14, 1993; Mazur et al., U.S. Patent No.
5,591,836,
assigned to The Procter & Gamble Co., issued January 7, 1997; Moreau et al.,
U.S.
Patent No. 5,843,499, assigned to United States of America, issued December 1,
1998;
Miettenen et al., U.S. Patent No. 5,958,913, assigned to Raisio Benecol Ltd.,
issued
September 28, 1999; Karppanen et al., WO 98/28990, assigned to Pharmaconsult,
published July 9, 1998; Shirakawa et al., EP 0,289,636, published November 9,
1988;
Ko, WO 94/18225, assigned to Du Pont Merck Pharmaceutical, published August
18,
1994; Festo, WO 95108342, assigned to Inpharma S.A., published March 30, 1995;
Ritter et al., WO 97/42830, assigned to Unilever PLC, published November 20,
1997;
Van AmeronAen et al., WO 98/01126, assigned to Unilever PLC, published January
15,
1998; and Wester et al., WO 98/06405, assigned to Raision Tehtaat, published
February 19, 1998. Any of the sterols and stanols described in the foregoing
references, as well as those commonly known in the art, may be included within
the
second component of the present compositions.
Thus, the term "sterol" as used herein can include natural or synthetic plant
or
animal sterols or triterpenes. This includes the phytosterols and the
mycosterols as well
as cholesterol, however it is preferred herein that cholesterol itself is not
utilized. For a
more detailed discussion of sterols see, for example, Nes, W.D., Parish, E.J.,
Eds.,
"Analysis of Sterols and Other Biologically Significant Steroids", Academic
Press, Inc.
(1989). Non-limiting examples of preferred sterols include diosgenin,
stigmastanol,
tigogenin, ~-sitosterol, ~-sitosterol, stigmasterol, ergosterol, campesterol,
oleanoic
acids, soyasapogenols, protoascigenin, togenols, protoparaxadiols,
protopanaxadiols,
~-amyrin, 0-amyrin, lupeol, butyrospermol, germanicol, 4-desmethylsterols, 4-
9
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
monomethylsterols, and 4,4'-dimethylsterols. Other non-limiting examples of
sterols for
use herein include 7-dehydrocholesterol, 22-dehydrocholesterol, 24-
dehydrocholesterol,
zymosterol, ~'-cholesterol, cerebrosterol, 22-~-oxycholesterol, 22-
dihydroerogosterol,
neospongosterol, cerebisterol, corbisterol, focosterol, 0-spinasterol,
sargasterol, 7-
dehydrocryonasterol, poriferasterol, chondrillasterol, cryonasterol (~-
sitosterol), dihydro-
~-sitosterol, 14-dehydroergosterol, 24(28)-dehydroergosterol, ergosterol,
brassicasterol, 24-methylenecholesterol, ascosterol, episterol, fecosterol,
and 5-
dihydroergosterol.
It is particularly preferred herein that phytosterols, the stanols derived
therefrom
(referred to herein as phytostanols), and esters thereof are utilized herein.
The term
phytosterol is intended to mean unsaturated sterol alcohois and their mixtures
derived
from plants, as well as synthetically produced sterol alcohols and their
mixtures which
are either identical to those sterols found in nature, or having properties
which are
similar to those of naturally occurring sterols. As is well-known in the art,
phytosterols
(also commonly referred to as plant sterols) are natural components of, for
example,
vegetable fats and oils. As is also commonly understood, the saturated forms
of these
sterols (i.e., the forms derived therefrom) are stanols.
The most preferred phytosterols for use as the second component herein
include sitosterol (e.g., ~-sitosterol (24-ethyl-5~-cholestane-30-ol) and 5~-
sitosterols),
stigmasterol, and campesterol. Schematic drawings of these components are as
given
in S.P. Kochhar, "Influence of Processing on Sterols of Edible Vegetable
Oils", Prog.
Lipid Res., Vol. 22, pp. 161 - 188. For example, 0-sitosterol has the
following sfiructure:
HsC H3C
CH3 %'~ ~ CH3
... H
CH3 H ~~- CH3
H~ ~
HO
Preparation of such phytosterols is commonly known; for example, sitosterol
can
be obtained from wood and from refining vegetable oil, and normally comprises
also a
minor amount of other sterols, such as campesterol, stigmasterol, and various
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
avenasterols. Other suitable phytosterols for use herein include
brassicasterol and
22,23-dihydrobrassicasterol.
As described herein above, one or more stanols may be utilized as the second
component of the present compositions. Stanols are found in small amounts in
nature
in such products as wheat, rye, corn, and triticale. They can also easily be
produced by
hydrogenation of natural sterol mixtures such as vegetable oil-based sterol
mixtures or
commercially available wood sterols. The plant sterols thus obtained can be
converted
into stanols by well-known hydrogenation techniques such as those based on the
use
of a Pd/C catalyst (or other similar catalyst) in organic solvent. A wide
variety of
palladium catalysts and solvents are known to those of ordinary skill in the
art and such
catalysis can be used to hydrogenate the sterol for formation of the desired
stanol. For
example, D-sitostanol (24-ethyl-50-cholestane-3~-ol) may be prepared by
hydrogenation of ~-sitosterol in organic solvent.
Accordingly, any sterol, including the foregoing examples of sterols, may be
utilized to provide the desired stanol. Accordingly, non-limiting examples of
useful
stanols include the hydrogenation products of the sterols described herein.
The most
preferred stanols herein include stanols of the phytosterols, for example,
sitostanols
(e.g., 0-sitostanol and 5~-sitostanols), campestanol, 24-~-methyl cholestanol,
stigmastanol, clionastanol, and dihydrobrassicastanol. For example, four major
plant
stanols are campestanol, 22,23-dihydrobrassicastanol, 0-sitostanol, and
clionastanol,
which have the following structure:
HOC
CH3
HO
H
wherein R is -CH3 for campestanol and its epimer, 22,23-dihydrobrassicastanol
and
wherein R is -C2H5 for sitostanol and its epimer, clionastanol. Campestanol
and 22,23-
dihydrobrassicastanol differ only by their steric configuration at C24.
Similarly, sitostanol
and clionastanol differ only by their steric configuration at C24. Alternate
nomenclature
11
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
for clionastanol is (30, 50, 24S)-stigmast-San-3-ol; sitostanol is (3~, 5~,
24R)-stigmast-
5an-3-ol; campestanol is (3~, 5~, 24R)-ergost-San-3-ol; dihydrobrassicastanol
is (3~,
5~, 24S)-ergost-San-3-ol.
It is further understood by one of ordinary skill that sterols, stanols, or
their
blends, can be utilized to produce sterol esters and / or stanol esters
utilized in the
present invention. As described below, such sterol and / or stanol esters are
also
particularly useful in the compositions of the present invention.
The esters of sterols and stanols are readily prepared by one of ordinary
skill in
the art. Utilization of sterol and / or stanol esters is particularly
preferred herein for
encapsulation of the first component. Such encapsulation is particularly
useful wherein
a sterol and / or stanol ester is used alone or, as particularly preferred, as
a mixture with
at least one sterol or stanol. It has been discovered that such encapsulation,
which is
described further herein below, surprisingly diminishes or removes the
unpalatable
flavor associated with the first component. Additionally, sustained delivery
of the first
component is accomplished through such encapsulation, providing enhanced and
prolonged bioavailability of the first component to provide the requisite
health benefits.
Furthermore, as stated above, use of the sterol and / or stanol ester provides
unique
health benefits as well.
The sterols and stanols herein may be esterified by any means utilizing any
appropriate precursor, for example, phenolic acids such as ferulic acid,
coumaric acid,
caffeic acid, and cinnamic acid. Other suitable acids include, for example,
citric acid,
lactic acid, oxalic acid, and malefic acid. However, for cholesterol-lowering
effects, and
other associated health benefits, fatty acid esterification is preferred. For
example,
mixtures of the fatty acids of any vegetable oil can be used. One example is a
mixture
of rapeseed oil and rapeseed oil fatty acid methyl ester. The preferred fatty
acids
useful herein are selected from saturated straight chain tatty acids,
saturated branched
chain fatty acids, and unsaturated fatty acids. The carbon chain length of the
fatty acid
useful in the present invention is preferably from 2 to about 24, more
preferably from
about 12 to about 24, even more preferably from about 16 to about 20, and most
preferably about 18.
Suitable examples of fatty acids useful for esterification herein include, for
example, valeric acid, isovaleric acid, sorbic acid, isocaproic acid, lauric
acid, myristic
acid, palmitic acid, stearic acid, arachidic acid, behenic acid, hexacosanoic
acid,
12
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
octacosanoic acid, pentadecanoic acid, heptadecanoic acid nonadecanoic acid,
tricosanoic acid, petacosanoic acid, decenylic acid, undecenylic acid,
dodecenylic acid,
oleic acid, linoleic acid, linolenic acid, arachidonic acid, erucic acid,
acetic acid,
propionic acid, butyric acid, caproic acid, caprylic acid, and capric acid.
More preferred
fatty acids include lauric acid, palmitic acid, stearic acid, arachidic acid,
behenic acid,
oleic acid, cetoleic acid, erucic acid, elaidic acid, linoleic acid, and
linolenic acid.
Additionally, fatty acid mixtures may be utilized, for example, mixtures of
rice bran oil,
sunflower oil, safflower oil, rapeseed oil, linseed oil, linola oil, and / or
soybean oil may
be utilized.
Such fatty acids may be utilized to provide the sterol and / or stanol fatty
acid
ester. Again, any of the foregoing sterols and stanols may be utilized, with
the
preferred limitations for sterols and stanols (e.g., phytosterols and stanols
derived
therefrom) being applicable for the fatty acid esters as well. For example,
sitostanol
fatty acid esters (e.g., ~-sitostanol fatty acid esters) are particularly
preferred for use
herein. Non-limiting examples of fatty acid esters include sitosterol acetate,
sitosterol
oleate, and stigmasterol oleate. Of course, the corresponding sitostanol fatty
acid
esters may also be utilized, e.g., sitostanol acetate, sitostanol oleate, and
sitgmastanol
oleate.
Other non-limiting examples of fatty acid sterol and stanol esters include
ergosta-5,7-lien-3-ol-9-hexadecenoate; (ergosta-5,7-dienylpalmitoleate);
ergosta-8,22-
dien-3-ol-14-methyl-4,9-octadecenoate; (14-~-methylergosta-8,22-dienyloleate);
lanost-
8-en-3-ol-9-octadecenoate; (dihydrolanosterol-oleate); ergost-5-en-3-ol-
9,12,15-
octadecatrienoate; dihydrobrassicasteryl-linolenate; ergost-5-en-3-ol-9,12-
octadecadienoate; ergost-5-en-3-ol-9-octadecenoate; dihydrobrassicasteryl-
oleate;
ergosta-7,24 (28)-dien-3-ol-4-methyl-9-octadecenoate; gramisteryl-oleate;
stigmasta-
8,24 (28)-dien-3-ol-9,12-octadecadienoate; 0' - avenasteryl-linoleate; ergosta-
7,24
(28)-dien-3-ol-4-methyl-9,12-octadecadienoate; gramisteryl-linoleate; stigmast-
24 (28)-
en-3-ol-9,12-octadecadienoate; ergosta-5,22-dien-3-ol-4,23-dimethyl-9-
octadecenoate;
ergostan-3-ol-4-methyl-9-octadecenoate; 5~-stigmastan-3~-ol-linolenate; 50-
stigmastan-30-ol-oleate; stigmastan-3-ol-9,12-octadecadienoate; 5~-stigmastan-
30-ol-
linoleate; 22-dihydrospinasteryl-linoleate; ergosta-5,7,22-trien-3-ol-9,12-
octa-
decadienoate; ergosterol-linoleate; stigmasta-5,24 (28)-dien-3-ol-9-
octadecenoate;
stigmasta-5,24 (28)-3-0l-9,12-octadecadienoate; stigmasta-5-en-3-ol-5,8,11,14-
13
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
eicosatetraenoate; ~-sitosterol-arachidonate; ergost-5-en-3-ol-5,8,11,14-
eicosatetraenoate; stigmasta-7,24 (28)-dien-3-ol-4-methyl-9,12-
octadecadienoate;
ergost-7-en-3-ol-9,12,15-octadecatrienoate; ergost-5-en-3-ol-9,12,15-
octadecatrienoate; campesteryl-linolenate; ergostan-3-ol-9,12-
octadecadienoate;
ergosta 5,24 (28)-dien-3-ol-9-hexadecenoate; ergosta-5,22-dien-3-ol-
octadecenoate;
brassicasteryl-oleate; lathosteryl-oleate; lanosta-8,24-dien-3-ol-9-
octadecenoate;
lanosterol-oleate; stigmasta-5,24(28)-dien-3-ol-9-octadecenoate; fucosteryl-
oleate;
desmosteryl-oleate; ergost-5-en-3-ol-12-octadecadienoate; campesteryl-
linoleate;
ergosta-5,22-dien-3-ol-9-octadecenoate; ergost-22-en-3-ol-9-hexadecenoate;
ergosta-
5,22-dien-3-ol-9-hexadecenoate; ergosta-5,22-dien-3-ol-9,12-octadecadienoate;
brassicasteryl-linoleate; ergosta-7,24(28)-dien-3-ol-9,12-octadecadienoate;
stigmasta-
5,22-dien-3-ol-9,12,15-octadecatrienoate; stigmasterol-linolenate; stigmasta-
5,22-dien-
3-0l-9,12-octadecadienoate; stigmasterol-linoleate; zymosteryl-oleate; ergost-
5-en-3-ol-
9-octadecenoate; campesteryl-oleate; ergosta-5,7,22-trien-3-ol-9-
hexadecenoate;
ergosterol-9-hexadecenoate; 50-stigmasta-7,22-dien-3~-ol-oleate; ~-spinasterol-
oleate; ergosta-5,7,22-trien-3-ol-9-octadecenoate; ergosterol-oleate; stigmast-
5-en-3-ol-
9-octadecenoate; D-sitosterol-oleate; stigmast-5-en-3-ol-9,12-
octadecadienoate; ~-
sitosterol-linoleate; stigmast-5-en-3-ol-9,12,15-octadecatrienoate; ~-
sitosterol-
linolenate; 0-sitosterol-undecenoate; ~-sitosterol-lauroylate; ~-sitosterol-
palmitate;
stigmasterol-undecenoate; stigmasterol-lauroylate; stigmasterol-palmitate; ~-
sitostanol-
oleate; 0-sitostanol-linoleate; ~-sitostanol-linolenate; 0-sitosterol-oleate;
5~-
stigmastan-3~-ol-oleate; 5~-stigmastan-3~-ol-linolenate; 100-ergosta-5,7,22-
trien-3~-
ol-linoleate; stigmast-5-en 3-ol-dodecenoate; ~-sitosterol-dodecenoate);
ergost-5-en-3-
ol-dodecenoate; campesteryl-dodecenoate; stigmasterol-dodecenoate; and 0-
sitosterol-dodecenoate.
Preparation of such fatty acid sterols and / or stanols are well-known to one
of
ordinary skill in the art. For example, Van Amerongen et al., WO 98/01126,
assigned to
Unilever PLG, published January 15, 1998, describes processes for the
manufacture of
a mixture of fatty acid esters comprising hydrolyzing a sterol ester or a
mixture of sterol
esters and esterifying the so obtained free sterols with particular fatty
acids. Preferably,
the conditions of the esterification reaction are chosen such that at least 50
wt%,
preferably at least 75 wt%, and most preferably from 90-100 wt% of the sterols
and / or
stanols are esterified. Other methods are disclosed in various references, for
example,
14
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Miettenen et al., U.S. Patent No. 5,958,913, assigned to Raisio Benecol Ltd.,
issued
September 28, 1999, which briefly describes esterification at a temperature of
90 °C to
120 °C under a vacuum of 5 to 15 mm Hg and using a catalyst such as
sodium ethylate.
Polyol Fatty Acid Pol esters
As an alternative to the sterols, stanols, and / or esters thereof, the second
component may be selected from polyol fatty acid polyesters. Polyol fatty acid
polyesters, and methods of their synthesis, are commonly known to provide no-
fat or
reduced calorie foods. Such polyol fatty acid polyesters are disclosed in, for
example,
Fulcher, U.S. Patent No. 4,582,927, issued April 15, 1986 (fatty esters of
malonic acid),
Vo~~enhein, U.S. Patent No. 4,582,715, issued April 15, 1986 (~-acetylated
triglycerides), Whyte, U.S. Patent No. 3,579,548, and issued May 18, 1991
(trigiycerides of ~-branched chain carboxylic acids). Other references which
describe
useful polyol fatty acid polyesters include Letton et al., U.S. Patent No.
5,306,514,
issued April 26, 1994; Letton et al., U.S. Patent No. 5,306,515, issued April
26, 1994;
Johnston et al., U.S. Patent No. 5,451,416, issued September 19, 1995; and
Elsen et
al., U.S. Patent No. 5,422,131, issued June 6, 1995. Polyol fatty acid
polyesters may
also be utilized in combination with, e.g., triglycerides, to provide low-fat
foods. For
example, Seiden et al., U.S. Patent No. 5,419,925, issued May 30, 1995
describes
reduced calorie fat compositions which contain combinations of polyol fatty
acid
polyesters and certain reduced calorie triglycerides.
The present inventors have surprisingly discovered that the first component,
as
described herein, may be combined with a polyol fatty acid polyester to
provide several
unexpected benefits. For example, it is known that certain polyol fatty acid
polyesters
are sensitive to oxidation and may oxidatively decompose under certain
conditions.
However, the present inventors have discovered that inclusion of the first
component
reduces susceptibility of the polyol fatty acid polyester to 'oxidative
decomposition. As
an additional unexpected benefit, the polyol fatty acid polyester diminishes
and / or
removes the undesirable flavor typically associated with the first component
of the
composition. Accordingly, as discovered herein, use of a polyol fatty acid
polyester as
the second component provides several unexpected advantages which have not
been
previously recognized.
Preferred among the polyol fatty acid polyesters are sucrose polyesters (i.e.,
sucrose in which at least four of the eight hydroxyl groups are esterified
with a fatty
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
acid). Sucrose polyester is a nondigestible fat which as been utilized in a
variety of
food compositions to provide non-fat foods. Such sucrose polyesters are
described in,
for example, the foregoing references. Particularly preferred sucrose
polyesters are
those sold under the trade name OLEANT"" and l or OLESTRAT"", by Procter &
Gamble
Co., Cincinnati, OH.
Flowable non-digestable polyol fatty acid polyesters, including sucrose
polyesters, are also particularly preferred herein. Flowable non-digestable
polyol fatty
acid polyesters and processes for making such polyol fatty acid polyesters are
disclosed in Cerreta et al., U.S. Patent Application Serial No. 08/844,590,
filed April 21,
1997.
Kits of the Present Invention
The present invention further relates to kits comprising a composition as
described herein and information that use of the composition provides
treatment
against general health benefits. Such general health benefits include, but are
not
limited to, cardiovascular benefits, including lowering cholesterol in the
consumer,
treating, preventing, and / or inhibiting heart disease (e.g.,
atherosclerosis, restenosis,
thrombosis) and, for example, treating other conditions such as
hypercholesterolemia,
hypertension, poor circulation, and other complications associated with
diabetes.
Additionally, the kit may comprise information that use of the
compound/composition
provides an organoleptic benefit, for example acceptable (e.g., good) flavor.
The information provided within the kit may for example, be oral information
disseminated as part of the kit, but is preferably written information. Such
written
information is typically present on packaging associated with the composition
(e.g., a
label present on a package containing the composition or package insert
included
within the kit). As used herein, "written" means through words, pictures,
symbols, and /
or other visible information. Such information need not utilize the actual
words but
rather use of pictures, symbols, and the like conveying the same or similar
meaning are
contemplated within the scope of this invention. Such information may also
include
information about general health benefits and reasons for which such health,
and
particularly treatment against certain disease states (including the
aforementioned
disease states), is important for the user.
16
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Methods of the Present Invention
The present invention also encompasses methods for providing certain health
benefits, particularly, lowering serum cholesterol or treating other
cardiovascular
problems or diseases (as set forth herein) comprising systemically (generally,
orally)
administering to a mammal (preferably, a human) successive therapeutically
effective
doses of the present compositions. Such methods include treating, preventing,
and l or
inhibiting (collectively referred to herein as treating) one or more of the
following:
cardiovascular problems including, but not limited to, atherosclerosis,
restenosis,
thrombosis, hypercholesterolemia, hypertension, diabetes, vascular
dysfunction, and
poor circulation, and other problems such as shock. Preferred methods herein
include
treatment of one or more of atherosclerosis, hypercholesterolemia,
hypertension,
diabetes, and poor circulation.
In accordance with the methods of the present invention, a present composition
is administered to a mammal, preferably a human. Preferably such
administration is
oral. As used herein, the term "oral administration" (or the like) with
respect to the
mammal (preferably, human) means that the mammal ingests or is directed to
ingest
(preferably, for the purpose of treatment of one or more of the various health
problems
described herein) one or more compositions of the present invention. Wherein
the
mammal is directed to ingest one or more of the compositions, such direction
may be
that which instructs and / or informs the user that use of the composition may
and / or
will provide treatment for the particular health problem of concern. For
example, such
direction may be oral direction (e.g., through oral instruction from, for
example, a
physician, sales professional or organization, and / or radio or television
media (i.e.,
advertisement) or written direction (e.g., through written direction from, for
example, a
physician or other medical professional (e.g., scripts), sales professional or
organization
(e.g., through, for example, marketing brochures, pamphlets, or other
instructive
paraphernalia), written media (e.g., Internet, electronic mail, or other
computer-related
media), and / or packaging associated with the composition (e.g., a label
present on a
package containing the composition). As used herein, "written" means through
words,
pictures, symbols, and / or other visible descriptors.
Administration of the present compositions may be via any systemic method,
however, such administration is preferably oral. Typically such administration
is at least
once monthly, but preferably weekly, and most preferably daily. Preferred
dosages of
17
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
the present compositions will vary. As one of ordinary skill will recognize
such
variations are largely dependent upon factors such as age, gender, weight, and
health
state of the consumer. However, it is often preferred that from about 0.05
grams to
about 50 grams of the first component is administered daily in such
composition. More
preferably, from about 0.01 grams to about 20 grams, even more preferably from
about
0.1 gram to about 10 grams, and most preferably from about 0.5 grams to about
6
grams of the first component is administered daily in such composition.
Additionally, as
a daily dose, typically from about 0.01 grams to about 50 grams of the second
component is administered daily. More preferably, from about 0.05 grams to
about 20
grams, even more preferably from about 0.1 gram to about 6 grams, and most
preferably from about 0.2 grams to about 4 grams of the second component is
administered daily.
Method of Making the Present Compositions
In accordance with the present invention, the mixture of the first component
and
the second component results in a thorough coating of the first component (L-
arginine,
salt, peptide, or pro-form thereof) which, as has been surprisingly discovered
herein,
diminishes or removes the strong, bitter, and / or fishy flavor
characteristics of L-
arginine. Several methods, including simple mixture of the first and second
components will be well-known in the art. However, for convenience, the
following is a
non-limiting example of a method of making the present compositions.
The particle size of the L-arginine (or salt, polypeptide, or pro-form
thereof)
should be reduced to minimize the perception of arginine particles in the
mouth, when
consuming the finished product. Typically such particle size will be less than
about 100
microns. The coating material is a second component of the present invention,
as has
been described herein. The coating material may be mixed with one or more
triglycerides, such as ~-3 fatty acids, to make the coating materials more
plastic or
deformable, which in turn provides enhanced texture of the final product.
The solvent system utilized is preferably an azeotropic mixture such that
during
the drying process solvent ratios are relatively maintained; such maintenance
will
enhance the solubility of the coated material. It has further been found that
using the
azeotropic mixture of solvents results in a uniform smooth thin film of coated
material.
Additionally, it has been discovered that wherein the azeotropic solvent
system is not
18
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
used the coated material can precipitate. Such precipitate will typically
cause the
coating to be lumpy and granular and (resulting in multiple particles sticking
together)
rather than smooth and uniform. It is important to avoid such precipitate
because the
precipitate can cause enhanced perceptibility of unacceptable flavor. Actual
coating of
the first component herein by the second component herein is described in the
following non-limiting examples. Variations of the following will be well-
known to one of
ordinary skill with the benefit of the present disclosure.
Example 1
A composition is prepared as follows having 40% phytosterol and
60°l° L-
arginine. Phytosterols (333 grams, commerically available from ADM, Decatur,
IIL) are
coated onto L-arginine using a solvent system of hexane and ethanol. A mixture
of
17% phytosterol, 69% hexane, and 21 % ethanol is heated to 55 °C. L-
arginine (about
500 grams) is loaded into a lab model Lakso Wurster Coater having a 4-inch by
6-inch
bowl (Model 101, commercially available from Lakso Co., Leominster, Ma). A 1l4
J-type
two-fluid nozzle having a fluid capacity of 20/50, air capacity of 70
manufactured by
Spraying Systems Co., Wheaton, III, is used to spray the phytosterol mixture.
The gap
between the Wurster insert and the distributor plate is adjusted depending
upon the
particle size of the L-arginine. A peristaltic pump is used to pump the
phytosterol
mixture to the nozzle. The mixture is circulated through the pumping system.
The
phytosterol solution should be adequately mixed by using a stir bar. The air
flow to the
unit is started. The air flow is adjusted until the bed is fluidizing
correctly. The humidity
of the inlet air to the bed is adjusted if necessary. The inlet air
temperature is adjusted
between about 100 °C and about 140 °C. The mixture is fluidized
at the desired air flow.
The blowback to the filter bags is turned on. The air pressure to the nozzle
is adjusted
to between about 20 and about 24 psi. When the desired bed temperature is
achieved,
the flow of the phytosterol mixture to the nozzle block is started. When all
of the
phytosterol mixture is sprayed, the bed is dried under ambient temperature and
pressure for about 10 minutes before turning off to allow the resulting
composition to
harden. After all appropriate settings are turned off, the bowl is unclamped
and the
phytosterol-coated L-arginine is removed.
Example 2
19
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Example 1 is repeated with use of 17% phytosterol and 83% L-arginine to
provide a composition providing substantially similar results.
Example 3
Example 1 is repeated with use of 60% phytosterol and 40% L-arginine to
provide a
composition providing substantially similar results.
Example 4
Example 1 is repeated with use of 23% phytosterol and 77% L-arginine to
provide a
composition providing substantially similar results. The coating material is a
mixture of
33% stigmasterol and 67% sitosterols, both supplied by Sigma Chemical Co., St.
Louis,
MO, in solvent system. The coating material contains 15% sterol and 85%
solvent
system.
Example 5
Example 1 is repeated with use of 35% stigmasterol (supplied by Sigma
Chemical Co., St. Louis, MO) and 65% L-arginine to provide a composition
providing
substantially similar results.
Use of the Present Co J~ositions and Kits
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 l or
inhibiting heart disease (e.g., atherosclerosis, restenosis, thrombosis) and,
for~example,
treating other conditions such as hypertension, poor circulation, and
complications
associated with diabetes. The compositions are useful in a wide variety of
finished
products, including pharmaceutical, food, and beverage products.
Preferred herein is use of the present compositions in food products,
including
those envisioned for use as a dietary supplement such as a health bar. In a
preferred
embodiment of the present invention, the compositions is in the form of a
health bar.
As non-limiting examples, 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, but are not limited to,
cakes,
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
brownies, muffins, bar cookies, health bars, wafers, biscuits, pastries, pies,
pie crusts,
and cookies, including sandwich cookies and chocolate chip cookies,
particularly the
storage-stable dual-textured cookies described in Hong et al., U.S. Pat. No.
4,455,333.
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, baked farinaceous
snack
foods, and other baked salted snacks.
As stated, health bars are a particularly preferred embodiment of the present
invention. The compounds can be incorporated into health bars, such as those
described in Greenberg et al., U.S. Patent No. 5,780,039. The foregoing doses
of the
present compounds may be included in the advantageous health bars according to
the
present invention.
In addition to their uses in baked goods, 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, but are not limited to, 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 are selected from margarines, butter,
dressings and
spreads.
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, including vegetable protein-based meat analog products, sauces,
gravies, and dairy products such as milkshakes, milk products, coffee
whiteners, and
cheese products.
The compounds described herein are also particularly useful in beverage
compositions. Such beverage compositions may be dilute water beverages (also
called
"near-water" beverages), milks, coffees, teas, colas, and fruit juices.
The compositions of the present invention may comprise one or more of the
following optional ingredients:
The isothiocyanate compound as described herein (optionally together with the
sorbate or benzoate preservative) is particularly useful in beverage products,
especially
21
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
dilute juice beverages, fortified beverages (e.g., calcium fortified
beverage), beverage
products containing tea solids (i.e., teas), and beverages containing milk
solids. The
isothiocyanate compound is most preferably present in the aqueous phase of the
beverage product for effective antimicrobial effect. Preferred beverage
products of the
present invention are those comprising a beverage member selected from the
group
consisting of water, fruit juice, tea solids, milk solids, fruit flavors,
botanical flavors, and
mixtures thereof. The beverage products herein are most preferably dilute
juice
beverages (particularly fruit juice beverages) and beverages containing tea
solids, and
beverage products comprising fruit juice and tea solids. Particularly
preferred beverage
products comprise both fruit juice and water. Other particularly preferred
beverage
products comprise both tea solids and water. In another preferred embodiment,
"near
water" (lightly flavored water) is utilized.
Various optional elements may be incorporated into the products and methods
of the present invention. Non-limiting examples of optional elements are as
follows:
Water
Water may be included in the compositions of the present invention,
particularly
wherein the compositions are beverage compositions. As used herein, the term
"water"
includes the total amount of water present in the composition. "Water"
includes water
from flavor agents, sugar syrups, and other sources, e.g., gum solutions.
Water of
hydration of, for example, calcium and other solids, is also included. Wherein
water is
included, water is preferably included at levels from about 0.1 % to about
99.999%,
more preferably from about 5% to about 99%, still more preferably from about
40% to
about 95%, even more preferably from about 50% to about 90%, and most
preferably
from about 70% to about 90%, by weight of the composition.
Beverage Emulsions
Dilute juice beverages of the present invention may optionally, but
preferably,
comprise from about 0.2% to about 5%, preferably from about 0.5% to about 3%,
and
most preferably from about 0.8% to about 2%, of a beverage emulsion. This
beverage
emulsion can be either a cloud emulsion or a flavor emulsion.
For cloud emulsions, the clouding agent can comprise one or more fats or oils
stabilized as an oil-in-water emulsion using a suitable food grade emulsifier.
Any of a
variety of fats or oils may be employed as the clouding agent, provided that
the fat or oil
is suitable for use in foods and l or beverages. Preferred are those fats and
oils that
22
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
have been refined, bleached and deodorized to remove off-flavors. Especially
suitable
for use as clouding agents are those fats that are organoleptically neutral.
These
include fats from the following sources: vegetable fats such as soybean, corn,
safflower, sunflower, cottonseed, canola, and rapeseed; nut fats such as
coconut,
palm, and palm kernel; and synthetic fats. See e.g., Kupper et al., U.S.
Patent No.
4,705,691, issued November 10, 1987, for suitable fat or oil clouding agents.
Any suitable food grade emulsifier can be used that can stabilize the fat or
oil
clouding agent as an oil-in-water emulsion. Suitable emulsifiers include gum
acacia,
modified food starches (e.g., alkenylsuccinate modified food starches),
anionic
polymers derived from cellulose (e.g., carboxymethylcellulose), gum ghatti,
modified
gum ghatti, xanthan gum, tragacanth gum, guar gum, locust bean gum, pectin,
and
mixtures thereof. See e.a., Ku~oper et al., U.S. Patent No. 4,705,691, issued
November
10, 1987. Modified starches treated to contain hydrophobic as well as
hydrophilic
groups, such as those described in Caldwell et al., U.S. Patent 2,661,349, are
preferred
emulsifiers for use as herein. Octenyl succinate (OCS) modified starches such
as those
described in Marotta et al., U.S. Patent 3,455,838 and Barndt et al., U.S.
Patent
4,460,617 are especially preferred emulsifiers.
The clouding agent can be combined with a weighting agent to provide a
beverage opacifier that imparts a total or partial opaque effect to the
beverage without
separating out and rising to the top. The beverage opacifier provides the
appearance
to the consumer of a juice-containing beverage. Any suitable weighting oil can
be
employed in the beverage opacifier. Typical weighting oils include brominated
vegetable oil, glycerol ester of wood rosin (ester gum), sucrose acetate
isobutyrate
(SAIB) and other sucrose esters, gum damar, colophony, gum elemi, or others
known to
those skilled in the art. Other suitable weighting agents include brominated
liquid polyol
polyesters which are nondigestible. See e.g., Brand et al., U.S. Patent
4,705,690,
issued November 10, 1987.
The cloudiopacifier emulsion is prepared by mixing the clouding agent with the
weighting agent (for opacifier emulsions), the emulsifier and water. The
emulsion
typically contains from about 0.1 % to about 25% clouding agent, from about 1
% to
about 20% weighting oil agent (in the case of opacifier emulsions), from about
1 % to
about 30% emulsifiers, and from about 25% to about 97.9% water (or quantum
satis).
23
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
The particle size of the water-insoluble components of the emulsion is reduced
by employing a suitable apparatus known in the art. Because the ability of
emulsifying
agents to hold oil in suspension is proportional to particle size, emulsions
of particles
with diameters of about 0.1 to about 3.0 microns are suitable. Preferably, the
particles
are about 2.0 microns or less in diameter. Most preferred is an emulsion in
which
substantially all the particles are 1.0 microns or less in diameter. The
particle size is
reduced by passing the mixture through an homogenizer, colloid mill or turbine-
type
agitator. Usually one or two passes is sufficient. See e.g., Kupper et al.,
U.S. Patent
4,705,691, issued November 10, 1987.
Flavor emulsions useful in beverage products of the present invention comprise
one or more suitable flavor oils, extracts, oleoresins, essential oils and the
like, known
in the art for use as flavorants in beverages. This component can also
comprise flavor
concentrates such as those derived from concentration of natural products such
as
fruits. Terpeneless citrus oils and essences can also be used herein. Examples
of
suitable flavors include, for example, fruit flavors such as orange, lemon,
lime and the
like, cola flavors, tea flavors, coffee flavors, chocolate flavors, dairy
flavors. These
flavors can be derived from natural sources such as essential oils and
extracts, or can
be synthetically prepared. The flavor emulsion typically comprises a blend of
various
flavors and can be employed in the form of an emulsion, alcoholic extract, or
spray
dried. The flavor emulsion can also include clouding agents, with or without
weighting
agents, as previously described. See e.~., Kupper et al., U.S. Patent
4,705,691, issued
November 10, 1987.
Flavor emulsions are typically prepared in the same manner as cloud/opacifier
emulsions by mixing one or more flavoring oils (from about 0.001 % to about
20%) with
an emulsifying agent (from about 1 % to about 30%) and water. (The oil
clouding
agents can also be present). Emulsions of particles with diameters of from
about 0.1 to
about 3.0 microns are suitable. Preferably, the particles are about 2.0
microns or less
in diameter. Most preferably, the particles are about 1.0 microns or less in
diameter.
The emulsifying agent coats the particularized flavor oil to aid in preventing
coalescence and in maintaining an appropriate dispersion. The viscosity and
specific
gravity of the flavor emulsion are regulated to be compatible with the
finished beverage.
See e.g., Kupper et al., U.S. Patent 4,705,691, issued November 10, 1987.
Flavor Agents
24
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
The compositions herein may optionally, but preferably, comprise one or more
flavor agents. Preferably, such flavor agents are included in the beverage
compositions
and are typically selected from fruit juice, tea solids, milk solids, fruit
flavors, botanical
flavors, and mixtures thereof. Wherein fruit juice is included, the beverages
of the
present invention can comprise from about 0.1 % to about 40%, preferably from
about
1 % to about 20%, more preferably from about 2% to about 10%, and most
preferably
from about 3% to about 6%, fruit juice. (As measured herein, the weight
percentage of
fruit juice is based on a single strength 2° to 16° Brix fruit
juice). The fruit juice can be
incorporated into the beverage as a puree, comminute, or as a single strength
or
concentrated juice. Especially preferred is incorporation of the fruit juice
as a
concentrate with a solids content (primarily as sugar solids) of from about
20° to about
80° Brix.
The fruit juice can be any citrus juice, non-citrus juice, or mixture thereof,
which
are known for use in dilute juice beverages. The juice can be derived from,
for
example, apple, cranberry, pear, peach, plum, apricot, nectarine, grape,
cherry, currant,
raspberry, gooseberry, elderberry, blackberry, blueberry, strawberry, lemon,
lime,
mandarin, orange, grapefruit, cupuacu, potato, tomato, lettuce, celery,
spinach,
cabbage, watercress, dandelion, rhubarb, carrot, beet, cucumber, pineapple,
coconut,
pomegranate, kiwi, mango, papaya, banana, watermelon, passion fruit,
tangerine, and
cantaloupe. Preferred juices are derived from apple, pear, lemon, lime,
mandarin,
grapefruit, cranberry, orange, strawberry, tangerine, grape, kiwi, pineapple,
passion
fruit, mango, guava, raspberry and cherry. Citrus juices, preferably
grapefruit, orange,
lemon, lime, and mandarin juices, as well as juices derived from mango, apple,
passion
fruit, and guava, as well as mixtures of these juices are most preferred.
Fruit flavors may also be utilized. As described above with respect to flavor
emulsions, fruit flavors may be derived from natural sources such as essential
oil and
extracts, or can be synthetically prepared. Fruit flavors may be derived from
fruits
through processing, particularly concentrating. Wherein fruit juices are
concentrated or
evaporated, the water which is removed or the condensate contains volatile
substances
which comprise the flavor of the fruit. Often, such flavor is added to a juice
concentrate
to enhance the flavor thereof. The condensate may also be used to flavor "near
waters" (lightly flavored water).
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Botanical flavors may also be utilized. As used herein, the term "botanical
flavor" refers to a flavor derived from parts of a plant other than the fruit;
i.e., derived
from nuts, bark, roots, and / or leaves. Also included within the term
"botanical flavor"
are synthetically prepared flavors made to simulate botanical flavors derived
from
natural sources. Botanical flavors can be derived from natural sources such as
essential oils and extracts, or can be synthetically prepared. Suitable
botanical flavors
include Jamaica, kola, marigold, chrysanthemum, chamomile, ginger, valerian,
yohimbe,
hops, eriodictyon, ginseng, bilberry, rice, red wine, mango, peony, lemon
balm, nut gall,
oak chip, lavender, walnut, gentiam, luo han guo, cinnamon, angelica, aloe,
agrimony,
yarrow and mixtures thereof.
Tannic acid or other similar acids can be used to provide an astringent taste
to
the beverage. From about 0.001 % to about 10% tannic acid is used. Other
flavor
enhancers, as well as flavorants such as chocolate and vanilla can also be
used.
Wherein tea solids are included, the beverages of the present invention can
comprise from about 0.01 % to about 1.2%, preferably from about 0.05% to about
0.8%,
by weight of the beverage product, of tea solids. The term "tea solids" as
used herein
means solids extracted from tea materials including those materials obtained
from the
genus Camellia including C. sinensis and C, assaimica, for instance, freshly
gathered
tea leaves, fresh green tea leaves that are dried immediately after gathering,
fresh
green tea leaves that have been heat treated before drying to inactivate any
enzymes
present, unfermented tea, instant green tea, and partially fermented tea
leaves. Green
tea materials are tea leaves, tea plant stems, and other plant materials that
are related
and which have not undergone substantial fermentation to create black teas.
Members
of the genus Phyllanthus, Catechu gambit and Uncaria family of tea plants can
also be
used. Mixtures of unfermented and partially fermented teas can be used.
Tea solids for use in beverages of the present invention can be obtained by
known and conventional tea solid extraction methods. A particularly preferred
source of
green tea solids can be obtained by the method described in Ekanayake et al.,
U.S.
Application Serial No. 08!606,907, filed February 26, 1996. Tea solids so
obtained will
typically comprise caffeine, theobromine, proteins, amino acids, minerals and
carbohydrates. Suitable beverages containing tea solids can be formulated
according
to Tsai et al., U.S. Patent 4,946,701, issued August 7, 1990. See also,
Ekanayake et
26
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
al., U.S. Patent 5,427,806, issued June 26, 1995, for a suitable sources of
green tea
solids for use in the present invention.
Beverages according to the present invention may also comprise milk solids.
These milk solids can be derived from various sources including whole milk,
skim milk,
condensed milk, and dried milk powder. As used herein, the term "milk" will be
used to
describe an aqueous dispersion of milk solids, such as fluid (whole or skim
milk) or non-
fat dry milk or condensed milk diluted with water. The amount of milk included
typically
ranges from about 5% to about 99.8%, preferably from about 5% to about 75%,
more
preferably from about 5% to about 40%, and most preferably from about 5% to
about
15%. The amount of non-fat milk solids correlating to these levels of milk
solids is in
the range of from about 0.5% to about 8.2%, from about 0.5% to about 6,2%,
from
about 0.5% to about 3,3%, and from about 0.5% to 1.2% of the beverage,
respectively.
Thickeners and Bulking Agents
Food and beverage compositions according to the present invention can further
comprise thickeners, including xanthan gum, carboxymethylcellulose,
carboxyethylcellulose, hydroxypropylcellulose, methylcellulose,
microcrystalline
cellulose, starches, dextrins, fermented whey, tofu, maltodextrins, polyols,
including
sugar alcohols (e.g., sorbitol and mannitol), carbohydrates (e.g., lactose),
propylene
glycol alginate, gellan gum, guar gum, pectin, tragacanth gum, gum acacia,
locust bean
gum, gum arabic, gelatin, as well as mixtures of these thickeners. These
thickeners are
typically included in the compositions of the present invention at levels up
to about
0.1 %, depending on the particular thickener involved and the viscosity
effects desired.
Sweeteners
The food and beverage compositions of the present invention can, and typically
will, contain an effective amount of one or more sweeteners, including
carbohydrate
sweeteners and natural and/or artificial no/low calorie sweeteners. The amount
of the
sweetener used in the compositions of the present invention typically depends
upon the
particular sweetener used and the sweetness intensity desired. For no/low
calorie
sweeteners, this amount varies depending upon the sweetness intensity of the
particular sweetener.
The compositions of the present invention can be sweetened with any of the
carbohydrate sweeteners, preferably monosaccharides and l or disaccharides.
Sweetened compositions, particularly beverages, will typically comprise from
about
27
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
0.1 % to about 20%, most preferably from about 6 to about 14%, sweetener.
These
sweeteners can be incorporated into the compositions in solid or liquid form
but are
typically, and preferably, incorporated as a syrup, most preferably as a
concentrated
syrup such as high fructose corn syrup. For purposes of preparing beverages of
the
present invention, these sugar sweeteners can be provided to some extent by
other
components of the beverage such as, for example, the fruit juice component and
l or
flavors.
Preferred sugar sweeteners for use in compositions of the present invention
are
sucrose, fructose, glucose, and mixtures thereof. Fructose can be obtained or
provided
as liquid fructose, high fructose corn syrup, dry fructose or fructose syrup,
but is
preferably provided as high fructose corn syrup. High fructose corn syrup
(HFCS) is
commercially available as HFCS-42, HFCS-55 and HFCS-90, which comprise 42%,
55% and 90%, respectively, by weight of the sugar solids therein, as fructose.
Other
naturally occurring sweeteners or their purified extracts, such as
glycyrrhizin, the protein
sweetener thaumatin, the juice of Luo Han Guo disclosed in, for example,
Fischer et al.,
U.S. Patent No. 5,433,965, issued July 18, 1995, and the like can also be used
in the
compositions of the present invention.
Suitable no/low calorie sweeteners include saccharin, cyclamates, L-aspartyl-L-
phenylalanine lower alkyl ester sweeteners (e.g., aspartame); L-aspartyl-D-
alanine
amides disclosed in Brennan et al., U.S. Patent No. 4,411,925; L-aspartyl-D-
serine
amides disclosed in Brennan et al., U.S. Patent 4,399,163; L-aspartyl-L-1-
hydroxymethylalkaneamide sweeteners disclosed in Brand, U.S. Patent No.
4,338,346;
L-aspartyl-1-hydroxyethyalkaneamide sweeteners disclosed in Rizzi, U.S. Patent
No.
4,423,029; L-aspartyl-D-phenylglycine ester and amide sweeteners disclosed in
Janusz,
European Patent Application 168,112, published January 15, 1986; N-[N-3,3-
dimethylbutyl)-L-D-aspartyl]-L-phenylalanine 1-methyl ester sweeteners
disclosed in
Gerlat et al., WO 99/30576, assigned to The Nutrasweet Co., published June 24,
1999;
alltame, thaumatin; dihydrochalcones; cyclamates; steviosides; glycyrrhizins,
synthetic
alkoxy aromatics, such as Dulcin and P-4000; sucrolose; suosan; miraculin;
monellin;
sorbitol, xylitol; talin; cyclohexylsulfamates; substituted imidazolines;
synthetic sulfamic
acids such as acesulfame, acesulfame-K and n-substituted sulfamic acids;
oximes such
as perilartine; rebaudioside-A; peptides such as aspartyl malonates and
succanilic
acids; dipeptides; amino acid based sweeteners such as gem-diaminoalkanes,
meta-
28
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
aminobenzoic acid, L-aminodicarboxylic acid alkanes, and amides of certain
alpha-
aminodicarboxylic acids and gem-diamines; and 3-hydroxy-4-alkyloxyphenyl
aliphatic
carboxylates or heterocyclic aromatic carboxylates; and the like and mixtures
thereof. A
particularly preferred low calorie sweetener is aspartame.
Coloring Agent
Small amounts of coloring agents may be utilized in the compositions of the
present invention. FD&C dyes (e.g., yellow #5, blue #2, red # 40) and / or
FD&C lakes
are preferably used. By adding the lakes to the other powdered ingredients,
all the
particles, in particular the colored iron compound, are completely and
uniformly colored
and a uniformly colored composition is attained. Preferred lake dyes which may
be
used in the present invention are the FDA-approved Lake, such as Lake red #40,
yellow
#6, blue #1, and the like. Additionally, a mixture of FD&C dyes or a FD&C lake
dye in
combination with other conventional food and food colorants may be used.
Riboflavin
and o-carotene may also be used. The exact amount of coloring agent used will
vary,
depending on the agents used and the intensity desired in the finished
product. The
amount can be readily determined by one skilled in the art. Generally, if
utilized, the
coloring agent should be present at a level of from about 0.0001 % to about
0.5%,
preferably from about 0.001 % to about 0.1 %, and most preferably from about
0.004%
to about 0.1 %, by weight of the composition.
Nutrients
The compositions herein (particularly the food and beverage compositions) can
be fortified with one or more nutrients, especially one or more vitamins,
minerals, and /
or amino acids. The U.S. Recommended Daily Intake (USRDI) for vitamins and
minerals are defined and set forth in the Recommended Daily Dietary Allowance-
Food
and Nutrition Board, National Academy of Sciences-National Research Council.
Any amino acid may be utilized herein, especially the naturally occurring
amino
acids. Preferred amino acids for inclusion herein are L-lysine and L-
carnitine,
particularly L-lysine.
Unless otherwise specified herein, wherein a given mineral is present in the
product, the product comprises at least about 1 %, preferably at least about
5%, more
preferably from about 10% to about 200%, even more preferably from about 40%
to
about 150%, and most preferably from about 60% to about 125% of the USRDI of
such
mineral. Unless othenivise specified herein, wherein a given vitamin is
present in the
29
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
product, the product comprises at least about 1 %, preferably at least about
5%, more
preferably from about 10% to about 200%, even more preferably from about 20%
to
about 150%, and most preferably from about 25% to about 120% of the USRDI of
such
vitamin.
Non-limiting examples of such vitamins and minerals include iron, zinc,
copper,
calcium, phosphorous, niacin, thiamin, folic acid, pantothenic acid, iodine,
vitamin A,
vitamin C, vitamin B~, vitamin B3, vitamin B6, vitamin B~2, vitamin D, vitamin
E, and
vitamin K. Preferably, wherein a vitamin or mineral is utilized the vitamin or
mineral is
selected from iron, zinc, calcium, niacin, thiamin, folic acid, iodine,
vitamin A, vitamin C,
vitamin B6, vitamin B~2, vitamin D, and vitamin E. A particularly preferred
mineral for
use herein is calcium.
Commercially available vitamin A sources may also be included in the present
compositions. Vitamin A can be provided, for example, as vitamin A palmitate
(retinol
palmitate) and / or as beta-carotene. The vitamin A may be in the form of, for
example,
an oil, beadlets or encapsulated. As used herein, "vitamin A" includes, but is
not limited
to, vitamin A, ~3-carotene, retinol palmitate, and retinol acetate. Wherein
vitamin A is
present in the compositions herein, the product comprises at least about 1 %,
preferably
at least about 5%, more preferably from about 10% to about 200%, even more
preferably from about 15% to about 150%, and most preferably from about 20% to
about 120% of the USRDI of such vitamin. Wherein vitamin A is present in the
products herein, it is especially preferred to include about 25% of the USRDI
of vitamin
A. The quantity of vitamin A to be added is dependent on processing conditions
and
the amount of vitamin A deliver desired after storage. Preferably, wherein
vitamin A is
included within the present compositions, the products comprise from about
0.0001 % to
about 0.2%, more preferably from about 0.0002% to about 0.12%, also preferably
from
about 0.0003% to about 0.1 %, even more preferably from about 0.0005% to about
0.08%, and most preferably from about 0.001 % to about 0.06% of vitamin A, by
weight
of the composition.
Commercially available sources of vitamin B2 (also known as riboflavin) may be
utilized in the present compositions. Wherein vitamin B2 is present in the
compositions
herein, the product comprises at least about 1 %, preferably at least about
5%, more
preferably from about 5% to about 200%, even more preferably from about 10% to
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
about 150%, and most preferably from about 10% to about 120% of the USRDI of
such
vitamin. Wherein vitamin B2 is present in the compositions herein, it is
especially
preferred to include from about 15% to about 35% of the USRDI of vitamin B2.
Commercially available sources of vitamin C can be used herein. Encapsulated
ascorbic acid and edible salts of ascorbic acid can also be used. Wherein
vitamin C is
present in the products herein, the product comprises at least about 1 %,
preferably at
least about 5%, more preferably from about 10% to about 200%, even more
preferably
from about 20% to about 150%, and most preferably from about 25% to about 120%
of
the USRDI of such vitamin. Wherein vitamin C is present in the compositions
herein, it
is especially preferred to include about 100% of the USRDI of vitamin C. The
quantity
of vitamin C to be added is dependent on processing conditions and the amount
of
vitamin C deliver desired after storage. Preferably, wherein vitamin C is
included within
the present compositions, the compositions comprise from about 0.005% to about
0.2%, more preferably from about 0.01 % to about 0.12%, also preferably from
about
0.02% to about 0.1 %, even more preferably from about 0.02% to about 0.08%,
and
most preferably from about 0.03% to about 0.06% of vitamin C, by weight of the
composition.
Commercial sources of iodine, preferably as an encapsulated iodine may be
utilized herein. ~ther sources of iodine include iodine-containing salts,
e.g., sodium
iodide, potassium iodide, potassium iodate, sodium iodate, or mixtures
thereof. These
salts may be encapsulated.
Nutritionally supplemental amounts of other vitamins which may be incorporated
herein include, but are not limited to, vitamins Bg and B12, folic acid,
niacin,
pantothenic acid, folic acid, vitamin D, and vitamin E. Wherein the
composition
comprises one of these vitamins, the product preferably comprises at least 5%,
preferably at least 25%, and most preferably at least 35% of the USRDI for
such
vitamin.
Minerals which may optionally be included in the composition herein are, for
example, magnesium, zinc, iodine, iron, and copper. Any soluble salt of these
minerals
suitable for inclusion edible products can be used, for example, magnesium
citrate,
magnesium gluconate, magnesium sulfate, zinc chloride, zinc sulfate, potassium
iodide,
copper sulfate, copper gluconate, and copper citrate.
31
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Calcium is a particularly preferred mineral for use in the present invention.
Preferred sources of calcium include, for example, amino acid chelated
calcium,
calcium carbonate, calcium oxide, calcium hydroxide, calcium sulfate, calcium
chloride,
calcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate,
calcium citrate, calcium malate, calcium titrate, calcium gluconate, calcium
realate,
calcium tantrate, and calcium lactate, and in particular calcium citrate-
malate. The form
of calcium citrate-malate is described in, e.g., Mehansho et al., U.S. Patent
No.
5,670,344, issued September 23, 1997; Diehl et al., U.S. Patent No. 5,612,026,
issued
March 18, 1997; Andon et al., U.S. Patent No. 5,571,441, issued November 5,
1996;
Meyer et al., U.S. Patent No. 5,474,793, issued December 12, 1995; Andon et
al., U.S.
Patent No. 5,468,506, issued November 21, 1995; Burkes et al., U.S. Patent No.
5,445,837, issued August 29, 1995; Dake et al., U.S. Patent No. 5,424,082,
issued
June 13, 1995; Burkes et al., U.S. Patent No. 5,422,128, issued June 6, 1995;
Burkes
et al., U.S. Patent No. 5,401,524, issued March 28, 1995; Zuniga et al., U.S.
Patent No.
5,389,387, issued February 14, 1995; Jacobs, U.S. Patent No. 5,314,919, issued
May
24, 1994; Saltman et al., U.S. Patent No. 5,232,709, issued August 3, 1993;
Camden et
al., U.S. Patent No. 5,225,221, issued July 6, 1993; Fox et al., U.S. Patent
No.
5,215,769, issued June 1, 1993; Fox et al., U.S. Patent No. 5,186,965, issued
February
16, 1993; Saltman et al., U.S. Patent No. 5,151,274, issued September 29,
1992;
Kochanowski, U.S. Patent No. 5,128,374, issued July 7, 1992; Mehansho et al.,
U.S.
Patent No. 5,118,513, issued June 2, 1992; Andon et al., U.S. Patent No.
5,108,761,
issued April 28, 1992; Mehansho et al., U.S. Patent No. 4,994,283, issued
February 19,
1991; Nakel et al., U.S. Patent No. 4,786,510, issued November 22, 1988; and
Nakel et
al., U.S. Patent No. 4,737,375, issued April 12, 1988. Preferred compositions
of the
present invention will comprise from about 0.01 % to about 0.5%, more
preferably from
about 0.03% to about 0.2%, even more preferably from about 0.05% to about
0.15%,
and most preferably from about 0.1 % to about 0.15% of calcium, by weight of
the
composition.
Iron may also be utilized in the compositions of the present invention.
Acceptable forms of iron are well-known in the art. The amount of iron
compound
incorporated into the ~ composition will vary widely depending upon the level
of
supplementation desired in the final product and the targeted consumer. Iron
fortified
compositions of the present invention typically contain from about 5% to about
100%,
32
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
preferably from about 15% to about 50%, and most preferably about 20% to about
40%
of the USRDI for iron.
Ferrous iron is typically better utilized by the body than ferric iron. Highly
bioavailable ferrous salts that can be used in the ingestible compositions of
the present
invention are ferrous sulfate, ferrous fumarate, ferrous succinate, ferrous
gluconate,
ferrous lactate, ferrous tartarate, ferrous citrate, ferrous amino acid
chelates, as well as
mixtures of these ferrous salts. While ferrous iron is typically more
bioavailable, certain
ferric salts can also provide highly bioavailable sources of iron. Highly
bioavailable
ferric salts that can be used in the food or beverage compositions of the
present
invention are ferric saccharate, ferric ammonium citrate, ferric citrate,
ferric sulfate, as
well as mixtures of these ferric salts. Combinations or mixtures of highly
bioavailable
ferrous and ferric salts can be used in these edible mixes and ready-to-serve
beverages. The preferred sources of highly bioavailable iron are ferrous
fumarate and
ferrous amino acid chelates.
Ferrous amino acid chelates particularly suitable as highly bioavailable iron
sources for use in the present invention are those having a ligand to metal
ratio of at
least 2:1. For example, suitable ferrous amino acid chelates having a ligand
to metal
mole ratio of two are those of formula:
Fe(L)2
where L is an alpha amino acid, dipeptide, tripeptide, or quadrapeptide
ligand. Thus, L
can be any ligand which is a naturally occurring alpha amino acid selected
from alanine,
arginine, asparagine, aspartic acid, cysteine, cystine, glutamine, glutamic
acid, glycine,
histidine, hydroxyproline, isoleucine, leucine, lysine, methionine, ornithine,
phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine;
or dipeptides,
tripeptides, or quadrapeptides formed by any combination of these alpha amino
acids.
See e.g., Ashmead et al., U.S. Patent No. 4,863,898, issued September 5, 1989;
Ashmead, U.S. Patent No. 4,830,716, issued May 16, 1989; and Ashmead, U.S.
Patent
No. 4,599,152, issued July 8, 1986, all of which are incorporated by
reference.
Particularly preferred ferrous amino acid chelates are those where the
reacting ligands
are glycine, lysine, and leucine. Most preferred is the ferrous amino acid
chelate sold
under the mark Ferrochel~ (Albion Laboratories, Salt Lake City, Utah) wherein
the
ligand is glycine.
33
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
In addition to these highly bioavailable ferrous and ferric salts, other
sources of
bioavailable iron can be included in the food and beverage compositions of the
present
invention. Other sources of iron particularly suitable for fortifying products
of the
present invention included certain iron-sugar-carboxylate complexes. In these
iron-
sugar-carboxylate complexes, the carboxylate provides the counterion for the
ferrous
(preferred) or ferric iron. The overall synthesis of these iron-sugar-
carboxylate
complexes involves the formation of a calcium-sugar moiety in aqueous media
(for
example, by reacting calcium hydroxide with a sugar, reacting the iron source
(such as
ferrous ammonium sulfate) with the calcium-sugar moiety in aqueous media to
provide
an iron-sugar moiety, and neutralizing the reaction system with a carboxylic
acid (the
"carboxylate counterion") to provide the desired iron-sugar- carboxylate
complex.
Sugars that can be used to prepare the calcium-sugar moiety include any of the
ingestible saccharidic materials, and mixtures thereof, such as glucose,
sucrose and
fructose, mannose, galactose, lactose, maltose, and the like, with sucrose and
fructose
being the more preferred. The carboxylic acid providing the "carboxylate
counterion"
can be any ingestible carboxylic acid such as citric acid, malic acid tartaric
acid, lactic
acid, succinic acid, propionic acid, etc., as well as mixtures of these acids.
These iron-sugar-carboxylate complexes can be prepared in the manner
described in, e.g., Nakel et al., U.S. Patent Nos. 4,786,510 and 4,786,518,
issued
November 22, 1988, both of which are incorporated by reference. These
materials are
referred to as "complexes", but they may exist in solution as complicated,
highly
hydrated, protected colloids; the term "complex" is used for the purpose of
simplicity.
Zinc may also be utilized in the compositions of the present invention.
Acceptable forms of zinc are well-known in the art. Zinc fortified products of
the present
invention typically contain from about 5% to about 100%, preferably from about
15% to
about 50%, and most preferably about 25% to about 45% of the USRDI for zinc.
The
zinc compounds which can be used in the present invention can be in any of the
commonly used forms such as, e.g., zinc sulfate, zinc chloride, zinc acetate,
zinc
gluconate, zinc ascorbate, zinc citrate, zinc aspartate, zinc picolinate,
amino acid
chelated zinc, and zinc oxide. Zinc gluconate and amino acid chelated zinc are
particularly preferred.
Carbonation Comaonent
34
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Carbon dioxide can be introduced into the water which is mixed with a beverage
syrup or into the dilute beverage after dilution to achieve carbonation. The
carbonated
beverage can be placed into a container, such as a bottle or can, and then
sealed. Any
conventional carbonation methodology may be utilized to make carbonated
beverage
products of this invention. The amount of carbon dioxide introduced into the
beverage
will depend upon the particular flavor system utilized and the amount of
carbonation
desired.
The compositions of the present invention, particularly the beverage
compositions, preferably have a pH of from about 2 to about 8, more preferably
from
about 2 to about 4.5, and most preferably from about 2.7 to about 4.2.
Beverage
acidity can be adjusted to and maintained within the requisite range by known
and
conventional methods, e.g., the use of food grade acid buffers. Typically,
beverage
acidity within the above recited ranges is a balance between maximum acidity
for
microbial inhibition and optimum acidity for the desired beverage flavor.
Non-Caloric or Reduced Calorie Fats
The compositions can be used in combination with non-caloric or reduced
calorie
fats, such as branched chain fatty acid triglycerides, triglycerol ethers,
polycarboxylic
acid esters, sucrose polyesters, sucrose polyethers, neopentyl alcohol esters,
silicone
oils/siloxanes, and dicarboxylic acid esters (particularly where the
composition is a food
composition). Other partial fat replacements useful in combination with the
fat materials
are medium chain triglycerides, highly esterified polyglycerol esters, acetin
fats,
polyoxyethylene esters, jojoba esters, mono/diglycerides of fatty acids, and
mono/diglycerides of short-chain dibasic acids.
Fiber Component
Similarly, food and beverage compositions can be made that combine the present
compositions with dietary fibers to achieve the combined benefits of each. By
"dietary
fiber" is meant complex carbohydrates resistant to digestion by mammalian
enzymes,
such as the carbohydrates found in plant cell walls and seaweed, and those
produced
by microbial fermentation. Examples of these complex carbohydrates are brans,
celluloses, hemicelluloses, pectins, gums and mucilages, seaweed extract, and
biosynthetic gums. Sources of the cellulosic fiber include vegetables, fruits,
seeds,
cereals, and man-made fibers (for example, by bacterial synthesis). Commercial
fibers
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
such as purified plant cellulose, or cellulose flour, can also be used.
Naturally occurring
fibers include fiber from whole citrus peel, citrus albedo, sugar beets,
citrus pulp and
vesicle solids, apples, apricots, and watermelon rinds.
These dietary fibers may be in a crude or purified form. The dietary fiber
used may
be of a single type (e.g., cellulose), a composite dietary fiber (e.g., citrus
albedo fiber
containing cellulose and pectin), or some combination of fibers (e.g.,
cellulose and a
gum). The fibers can be processed by methods known to the art.
Primarily due to the present compositions, the foods and beverages herein can
provide reduced serum cholesterol and thus reduced risk of heart disease.
Additionally,
the present compositions have acceptable organoleptic properties, particularly
flavor
and texture, despite the presence of L-arginine, polypeptides thereof, salts
thereof, and
pro-forms thereof.
Dietary foods can be made with the compositions to meet special dietary needs,
for example, of persons who are obese, diabetic, or hypercholesterolemic. The
present
compositions can be a major part of a low-fat, low-calorie, low-cholesterol
diet, and they
can be used alone or in combination with drug therapy, nutritional therapy, or
other
therapy. Combinations of food or beverage products made with the compositions
can
be used as part of a total dietary management regimen, based on one or more of
these
products, containing the compositions alone or in combination with one or more
of the
above-mentioned ingredients, to provide one or more of the above-mentioned
benefits.
This discussion of the composition uses, combinations, and benefits, is not
intended to be limiting or all-inclusive. It is contemplated that other
similar uses and
benefits can be found that will fall within the spirit and scope of this
invention.
Examples
The following examples are illustrative of uses of the present compositions.
Such examples are non-limiting illustrations and various modifications thereof
may be
made by one of ordinary skill in the art with the benefit of the present
disclosure.
Example 6
L-arginine is coated under conditions similar to those described herein above.
L-arginine in powdered form, and having a particle size of less than about 100
microns
is utilized. The L-arginine is coated with a mixture of a sterol and a sterol
ester or a
36
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
mixture of a stanol and a stanol ester. The ratio of sterol/stanol to ester is
adjusted to
provide a coated material with a malleable form. Sufficient coating is
deposited upon
the arginine particle to ensure flavor protection. The resulting coated
material is mixed
into a commercially available peanut butter preparation at a level equivalent
to 10
grams of coated material per 100 grams of peanut butter. Two and one half
grams of
peanut butter, containing approximately 0.175 grams of L-arginine, is applied
to two
crackers to form a sandwich. The sandwiches are sensory evaluated in the
laboratory
and exhibit no bitter or fishy off-flavors or aftertaste relative to control
peanut butter
sandwiches.
Example 7
In a manner similar to that described in Example 6, L-arginine coated with a
mixture of
sterol and sterol fatty acid ester or stanol and stanol fatty acid ester is
added to a
cheddar cheese preparation at a level of 10 grams per 75 grams of cheese
mixture.
Approximately 3 grams of the resulting mixture (containing approximately 0.176
grams
of L-arginine) was applied to commercially available crackers. The crackers
having the
mixture applied thereto are sensory evaluated in the laboratory and exhibit no
bitter or
fishy off-flavors or aftertaste relative to crackers having a control cheese
mixture
applied thereto.
Example 8
In a manner similar to that described in Example 6, L-arginine coated with a
mixture of
sterol and sterol fatty acid ester. A high ratio of sterol relative to sterol
ester is used to
prepare a coating with excellent thermal stability. The resulting coated L-
arginine is
added to prepared, commercially available sugar cookie mix at a ratio of 18
grams of
coated L-arginine to 510 grams of cookie mix. Eighteen cookies are prepared
according to known procedures, each containing approximately 0.5 grams of L-
arginine.
The resulting cookies retain their natural cookie flavor.
37
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Example 9
A fat-free health bar is prepared having the following composition:
Component Wt
Soy Protein Isolates 28
Fructose 30
High Fructose Corn Syrup 23.5
Raisins 6.8
Coated L-Arginine (Coated as described5
herein with a mixture of sterol
and sterol fatty
acid ester)
OIeanT"" (sucrose polyester, commercially6
available from Procter & Gamble
Co.,
Cincinnati, OH)
Cinnamon 0.5
Salt 0.1
Sodium Bicarbonate 0.1
The Sterol ester of L-arginine and OIeanT"" are pre-mixed prior to blending
with the
remainder of the dry ingredients and formed into bars. Other dried fruits, for
example,
cranberries, apricots, and the like may be substituted for the raisins. The
health bar is
ingested once daily for a period of 12 weeks as a supplement to a normal diet.
The
health bar is shown to reduce serum cholesterol levels after this 12 week
period.
38
CA 02395446 2002-06-20
WO 01/54686 PCT/USO1/02382
Example 10
A sports energy gel is prepared having the following composition:
Component Wt
Maltodextrin 59
Water 20
Fructose 12
Coated L-Arginine (Coated as described5
herein with a mixture of sterol
and sterol fatty
acid ester)
Citric Acid 3
Vitamin C 0.5
Vitamin E 0.1
Artificial Flavor 0.2
Sodium Benzoate 0.1
Potassium Sorbate 0.1
39
