U.S. patent application number 12/621312 was filed with the patent office on 2010-05-27 for protein hydrolysate and plant sterol containing composition for improving serum lipid profile and preventing atheroscleroris.
This patent application is currently assigned to RAISIO NUTRITION LTD.. Invention is credited to Paivi Kuusisto, Ingmar WESTER.
Application Number | 20100130401 12/621312 |
Document ID | / |
Family ID | 8562560 |
Filed Date | 2010-05-27 |
United States Patent
Application |
20100130401 |
Kind Code |
A1 |
WESTER; Ingmar ; et
al. |
May 27, 2010 |
PROTEIN HYDROLYSATE AND PLANT STEROL CONTAINING COMPOSITION FOR
IMPROVING SERUM LIPID PROFILE AND PREVENTING ATHEROSCLERORIS
Abstract
The invention relates to a terapeutical composition comprising a
protein hydrolysate and a plant sterol, wherein the weight ratio of
the plant sterol to the protein hydrolysate is from 1:0.02 to
1:150. The invention also relates to a terapeutical composition
comprising a protein hydrolysate and a synthetic emulsifier and/or
a lipid based grain fraction having emulsifying properties. Said
compositions can be used in a pharmaceutical, nutraceutical or food
product for improving serum lipid profile.
Inventors: |
WESTER; Ingmar; (Turku,
FI) ; Kuusisto; Paivi; (Naantali, FI) |
Correspondence
Address: |
ARENT FOX LLP
1050 CONNECTICUT AVENUE, N.W., SUITE 400
WASHINGTON
DC
20036
US
|
Assignee: |
RAISIO NUTRITION LTD.
Raisio
FI
|
Family ID: |
8562560 |
Appl. No.: |
12/621312 |
Filed: |
November 18, 2009 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10498519 |
Jan 10, 2005 |
|
|
|
PCT/FI2002/001053 |
Dec 20, 2002 |
|
|
|
12621312 |
|
|
|
|
Current U.S.
Class: |
514/1.1 |
Current CPC
Class: |
A23L 33/11 20160801;
A61K 38/01 20130101; A23V 2002/00 20130101; A23L 2/02 20130101;
A61P 3/04 20180101; A23L 27/60 20160801; A23C 11/103 20130101; A23C
19/093 20130101; A61P 9/10 20180101; A23C 9/13 20130101; A21D 2/14
20130101; A23L 33/18 20160801; A61P 3/06 20180101; A23L 7/126
20160801; A61P 9/00 20180101; A23C 19/0765 20130101; A23D 7/015
20130101; A23L 33/22 20160801; A23D 7/0056 20130101; A21D 2/268
20130101; A21D 2/16 20130101; A61K 38/01 20130101; A61K 2300/00
20130101; A23V 2002/00 20130101; A23V 2250/5488 20130101; A23V
2250/2136 20130101 |
Class at
Publication: |
514/2 |
International
Class: |
A61K 38/01 20060101
A61K038/01; A61P 3/06 20060101 A61P003/06 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 21, 2001 |
FI |
20012553 |
Claims
1. A method of lowering serum triglycerides in a subject,
comprising administering to the subject an oral therapeutical
composition comprising a protein hydrolysate and a plant sterol,
wherein the weight ratio of the plant sterol to the protein
hydrolysate is from 1:0.02 to 1:150.
2. The method according to claim 1, wherein the composition
comprises an additional component selected from the group
consisting of emulsifiers, fats, mineral salts and mixtures
thereof.
3. The method according to claim 1, wherein the plant sterol is
selected from the group consisting of free sterols, free stanols,
sterols in esterified form, stanols in esterified form and mixtures
thereof.
4. The method according to claim 1, wherein the protein hydrolysate
is derived from vegetable or animal origin, including soybean
protein, wheat protein, corn protein, oat protein, rye protein,
rapeseed protein, barley protein, milk protein and mixtures
thereof.
5. The method according to claim 2, wherein the emulsifier is
selected from the group consisting of monoglycerides, diglycerides,
monoglyceride derivatives, lecithins, modified lecithins,
polyglycerol esters, polyglycerol polyricinoleate, sorbitan esters,
polysorbates, propylene glycol esters, stearoyl lactylates,
diacetyl tartaric acid esters, diacetyl lactic acid esters, sucrose
esters and mixtures thereof.
6. The method according to claim 2, wherein the emulsifier has a
hydrophilic lipophilic balance (HLB) of at least 4.
7. The method according to claim 1, wherein the weight ratio of the
plant sterol to the protein hydrolysate is from 1:0.2 to 1:30.
8. The method according to claim 2, wherein the protein hydrolysate
is in the form of a protein hydrolysate/emulsifier complex.
9. The method according to claim 8, wherein the complex comprises
at least 5%.
10. The method according to claim 2, wherein the plant sterol is in
the form of a plant sterol/emulsifier complex or dissolved or
suspended in fat or a mixture of fat and an emulsifier.
11. The method according to claim 2, wherein the protein
hydrolysate and the plant sterol are in the form of a protein
hydrolysate/plant sterol/emulsifier complex.
12. The method according to claim 10, wherein the weight ratio of
the plant sterol to emulsifier is from 1:0.01 to 1:5.
13. The method according to claim 2, wherein the fat content is
from 0 to 80% of the weight of the complex or suspension or
solution.
14. A method of lowering serum triglycerides in a subject,
comprising administering to the subject an oral therapeutical
composition comprising a protein hydrolysate and at least one
emulsifier or a mixture of at least one emulsifier and a fat,
wherein the emulsifier is a synthetic emulsifier.
15. A method of lowering serum triglycerides in a subject,
comprising administering to the subject an oral therapeutical
composition comprising a protein hydrolysate and at least one
emulsifier or a mixture of at least one emulsifier and a fat,
wherein the emulsifier is a lipid based grain fraction having
emulsifying properties.
16. A method of lowering serum triglycerides in a subject,
comprising administering to the subject a food product comprising
at least one basic nutritional ingredient and an oral therapeutical
composition comprising a protein hydrolysate and a plant sterol,
wherein the weight ratio of the plant sterol to the protein
hydrolysate is from 1:0.02 to 1:150.
17. The method according to claim 16, wherein the food product is
in the form of a product selected from the group consisting of
bakery, confectionery, cereal, fermented cereal, snacks, beverage,
dairy, sauce, soup, meat, fish, poultry, egg, soy based, vegetable
oil based and ready mix products.
18. The method according to claim 16, wherein the plant sterol
content calculated as sterol equivalents is from 0.05 to 20 g per
100 g food product and the protein hydrolysate content is from
0.0002 to 20 g per 100 g food product.
28. The method according to claim 1, wherein the protein
hydrolysate is administrated at a rate of 0.1 to 60 g per day, and
the plant sterol is administrated at a rate of 0.4 to 5 g per day
calculated as sterol equivalents.
Description
FIELD OF INVENTION
[0001] The present invention relates to the field of nutrition and
health. Especially it concerns improved compositions for oral use
suitable for improving serum lipid profile including reducing serum
total and/or LDL cholesterol and/or apolipoprotein B levels.
BACKGROUND OF THE INVENTION
[0002] Cardiovascular disease (CVD) is one of the major causes of
death in western world. Elevated serum total and/or LDL (low
density lipoprotein) cholesterol and triacyl-glycerol levels, as
well as low ratio of HDL (high density lipoprotein) cholesterol to
LDL cholesterol, are some of the major risk factors for CVD.
Recently also serum levels of apolipoprotein B 100 (apo B) has been
shown to be a reliable CVD risk marker. In most developed countries
a substantial amount of the population have serum cholesterol
levels not within the recommended levels. One of the first steps in
improving the serum lipid profile is changes in life style
including changes in diet and exercise. It seems, however,
difficult to change dietary habits and to follow dietary
recommendations. Thus, there is a clear need for solutions beyond
regular diet and lifestyle changes by which serum lipid profile can
be improved.
[0003] Food products enriched with components having cholesterol
lowering effect beyond normal nutrition have been commercially
available for some time. Representative examples are food products
enriched with plant stanol or sterol fatty acid esters. Stanol
fatty acid esters and the cholesterol lowering effects thereof are
disclosed in U.S. Pat. No. 5,502,045 as well as a suitable method
for their preparation. Dietary intake of 2 to 3 g/day of plant
stanols is reported to lower serum LDL cholesterol levels in man up
to 14% without affecting HDL cholesterol levels. The amount of
atherogenic apolipoprotein B particles in blood is also decreased,
thus reducing the risk of CVD.
[0004] Some proteins have also been used to improve serum lipid
profile. Proteins that have been shown to lower serum total and/or
LDL cholesterol include e.g. soy protein (Anderson J. W., et al,
New Engl. J. Medicine 1995 333 (5) 276-282), whey protein (Nagaoka
et al., Agric. Biol. Chem. 1991 (55) 813-818) and wheat gluten (EP
0 790 060 A1). FDA has approved a health claim about the role of
soy protein in reducing the risk of CVD by lowering blood
cholesterol levels. In order to qualify for this health claim, a
food must contain at least 6.25 grams of soy protein per serving,
the amount being one-fourth of the effective level of 25 grams per
day, and be included in a diet low in saturated fat and
cholesterol.
[0005] EP 0 669 835 B1 describes a hypocholesterolemic dietary
composition containing soy protein and sitosterols, the sitosterols
and soy protein being of natural origin and having synergistic
cholesterol lowering effect. The composition is used for example as
tablets, capsules or syrups. Effective daily intakes of the
composition are not given Also EP 1 046 396 A2 describes a
composition containing soy protein, preferably soy protein isolate
or soy protein concentrate, and plant sterols having synergistic
cholesterol lowering effect. The synergistic effect is achieved,
when both the soy protein and plant sterols are administered in
amounts that would be sufficient to induce cholesterol-lowering
activity of each of the administered compound alone. WO 01/37681
describes a composition containing phytosterols and an isolated
water-soluble protein, such as soy protein or caseinate, and
optionally also an emulsifier. The composition can be utilized in
food products, where the composition improves mouthfeel and
stability of the sterol containing foods. No improved cholesterol
lowering effect compared to the effect achieved with phytosterols
alone is attributed to this composition.
[0006] U.S. Pat. No. 6,113,972 describes a phytosterol protein
complex, where the complex increases the bioavailability of the
phytosterols. No cholesterol lowering effect is attributed to the
protein part of this composition, but the protein functions only as
a carrier to the phytosterols. In fact, any fat binding protein can
be used in this complex and especially egg proteins are preferred.
A composition containing plant sterols/stanols and an emulsifier,
mixed into a protein-containing product, is described in WO
00/33669. The protein does not have any cholesterol lowering effect
in this composition either, but it aids in the formation of the
emulsion.
[0007] Protein hydrolysates have been shown to have greater serum
cholesterol lowering effect than the corresponding unhydrolyzed
proteins (Sugano et al., J. Nutr. 1990 (120) 977-985). The
cholesterol lowering effect of protein hydrolysates might be
attributed to certain peptides, for example such as found from
bovine milk .beta.-lactoglobulin by Nagaoka et al. (Biochem.
Biophys. Res. Commun., 2001, 281(1), 11-17), but this still remains
to be proven by well-controlled clinical and/or animal studies. EP
0 790 060 A1 discloses a protein hydrolysate/phospholipid complex
for improving lipid metabolism, where the complex contains 10% or
more bound phospholipids, especially lecithin or enzyme modified
lecithin. Lipid profile improving effect, especially
LDL-cholesterol lowering and HDL-cholesterol raising effect of
lecithin is well known in prior art (for example Childs et al.,
1981, Atherosclerosis 38, 217). In the complex according to EP 0
790 060 A1 certain bound phospholipids (lecithin or enzyme modified
lecithin) are believed to remarkably contribute to the lipid
metabolism improving effect of the complex.
[0008] The current literature and especially FDA's interim approval
of a health claim for plant sterols have increased the interest of
the food industry in supplementing foods with plant sterols.
Indeed, many such food items have recently been introduced into the
market. It is assumed that new plant sterol containing foods will
appear into the market rapidly.
[0009] The active launching of new plant sterol containing foods
has raised a concern that a part of the population may have higher
daily intake of plant sterols than what is needed to obtain an
optimal cholesterol lowering effect. Short-term high intake of
plant sterols has not been shown to be harmful. However, there is
no data on possible long-term side effects of ingestion of high
daily amounts (>3 g/day or higher) of plant sterols.
Furthermore, there is currently not a clear understanding of the
biological impact of the increased serum level and thus higher
systemic availability of plant sterols caused by increased intake
from sterol enriched foods, especially foods enriched with
unsaturated plant sterols.
[0010] There is a clear need for improved solutions by which the
serum lipid profile can be improved, e.g. elevated serum total
and/or LDL cholesterol levels reduced and/or HDL cholesterol levels
increased and/or triacylglycerol levels reduced and/or
apolipoprotein B levels reduced. Compositions based on combinations
of active ingredients having enhanced, additive or even synergistic
effects compared to the active ingredients alone are needed. It
would also be desirable to obtain meaningful and sustainable
improvements in the serum lipid profile with lower daily intake of
the individual active ingredients than what is obtainable with the
intakes currently used. Possible adverse effects of the active
ingredients such as possible impact on fat-soluble antioxidants and
other digestive discomforts could thus be reduced. Incorporation of
the active ingredients into a wider range of food products without
adverse effects on organoleptic properties is also needed. The
present invention provides compositions meeting these
requirements.
SUMMARY OF THE INVENTION
[0011] The present invention provides improved compositions for
improving serum lipid profile in humans and/or animals and a method
for improving serum lipid profile in humans and/or animals by
orally administering the composition. In one aspect the
compositions comprise combinations of protein hydrolysates and
plant sterols. The compositions may optionally comprise also
emulsifier(s) and/or fat and/or mineral salt. The compositions
comprise one or more protein hydrolysates in combination with one
or more plant sterols (sterols and/or stanols) in their free and/or
esterified forms, optionally in combination with one or more
emulsifiers and/or fat and/or mineral salt. Preferably the
emulsifier is used to produce a complex with protein hydrolysates
and/or sterols. In a second aspect the compositions comprise
combinations of protein hydrolysates and synthetic emulsifier(s).
The compositions may optionally comprise also fat and/or mineral
salt. In a third aspect of the invention the compositions comprise
combinations of protein hydrolysates and lipid based grain
fractions having emulsifying properties. The compositions may
optionally comprise also fat and/or mineral salt.
[0012] The compositions can be used as such or as pharmaceuticals
or nutraceuticals or most advantageously in food products.
Typically such an improved composition is a complex containing soy
protein hydrolysate, emulsifier and plant sterols, advantageously
used in food products for improving the serum lipid profile.
[0013] It was found that the combination of protein hydrolysates
and plant sterols, optionally with emulsifiers and/or fat and/or
mineral salt, further enhance the serum lipid profile improving
effect that is obtained by using the individual active ingredients
and even work synergistically in improving the overall serum total
lipid profile, for instance in reducing blood serum total and/or
LDL cholesterol levels, and have greater effect than the additive
effect that was expected. In addition some of the combinations,
especially those containing soy protein hydrolysate showed
beneficial effect on HDL cholesterol levels. Also the lowering of
serum triglyceride levels was shown to be surprising effective.
[0014] The present invention provides also means for further
improving of serum lipid profile by using combinations of the
active ingredients compared to the effects obtained when one of the
ingredients is used alone. For example using the compositions of
the present invention in combination with a healthy recommended
diet provides means by which serum LDL cholesterol level can be
reduced even by 25% or more. The present invention thus provides a
versatile non-drug approach to improve serum lipid profile, e.g. to
lower serum total and/or LDL cholesterol, especially in subjects
having serum lipid levels not within the recommended limits. The
present invention also provides combinations of the active
ingredients, especially those containing mineral salt and/or
specific protein hydrolysates, that have beneficial effect on blood
pressure. The combination of the active ingredients according to
the present invention thus reduces the risk of CVD beyond what is
obtainable by using only one of the active ingredients.
[0015] The present invention further provides means for minimizing
the daily intake of plant sterols in humans and still achieving a
similar serum lipid profile improving effect as obtained with
recommended daily intake of plant sterols from commercial plant
sterol enriched products. This is especially important as the daily
amount of plant sterols needed for obtaining a reduction in serum
total and/or LDL cholesterol can be reduced, thus preventing
potential over-consumption of plant sterols. Furthermore, the
commercial availability of plant sterols is limited. In addition
the commercial crude plant sterols are expensive due to their
limited availability, whereas the availability of raw materials for
protein hydrolysates or protein hydrolysate complexes usable
according to this invention is not restricted.
[0016] By the use of the compositions according to the present
invention the plant sterol supply can be more effectively utilized
with the aim of reducing the risk of CVD in subjects as well as in
the whole population. The present invention provides a way of
obtaining an enhanced serum lipid profile improvement or a similar
lipid profile improvement as obtained with plant sterols or protein
hydrolysate based ingredients alone, but especially with lower
daily intake of plant sterols and/or protein hydro-lysate. As
active ingredients based on protein hydrolysates are cheaper than
plant sterols the present invention also provides a way of
obtaining an optimal lipid profile improving effect more cost
effectively.
[0017] Compared to combinations of plant sterols and unhydrolyzed
proteins previously known in the art, the compositions of the
present invention comprising the combination of plant sterols and
protein hydrolysates, optionally with emulsifiers and/or fat and/or
mineral salt, have enhanced serum lipid profile improving effect
and can also be utilized in a wider range of food products and by a
wider range of consumers. In addition to having greater cholesterol
lowering effect compared to unhydrolyzed proteins, protein
hydrolysates are also less allergenic than unhydrolyzed proteins,
thus enabling wider range of consumers being able to benefit the
serum lipid profile improving effect. The compositions of the
present invention can also be more easily incorporated into food
products, such as beverages, without adverse effects on the texture
due to the reduced viscosity of some of the protein hydrolysates
compared to unhydrolyzed proteins.
[0018] A problem encountered with protein hydrolysates and their
incorporation into food products is often a pronounced bitter taste
of the hydrolysate. The bitter taste is the result of cleavage of
the proteins at amino acids with hydrophobic side chains, which
results in formation of peptides with exposed hydrophobic amino
acid residues. Hydrolysates may thus have an adverse effect on the
taste of the food product where they are incorporated. In addition
to protein hydrolysates, also some protein hydrolysate/phospholipid
complexes have bitter taste and can adversely affect the taste of
the products containing them. In addition to off-flavors, some
protein hydrolysate/phospholipid complexes, especially complexes
containing soy protein hydrolysate and lecithin or modified
lecithin, have also strong yellowish or brown colour and their use
in light coloured products, such as some dairy products, may thus
cause off-colours to the product.
[0019] The present invention by combining plant sterols and protein
hydrolysates, optionally with emulsifiers and/or fat and/or mineral
salt, in lipid profile improving products, such as foods, provides
a way of obtaining at least similar lipid profile improving effect,
especially cholesterol lowering effect, as can be obtained with
protein hydrolysates alone, but with lower protein hydrolysate
content and thus with less adverse effect on the taste of the food
products. Complexing protein hydrolysates either with synthetic
emulsifiers or with plant sterols or advantageously with both
emulsifiers and plant sterols also diminishes the unpleasant taste
of protein hydrolysates. The compositions according to the present
invention thus widen the range of food products in which the
protein hydrolysates can be incorporated to include also
mild-tasting ones and still achieving at least similar serum lipid
profile improving effect. The compositions of the present invention
also widen the range of food products in which protein
hydrolysate/emulsifier complexes can be incorporated to include
also light coloured products. The present invention thus also
provides a method to improve the sensory properties of lipid
profile improving foods containing protein hydrolysates or protein
hydrolysate/emulsifier complexes.
DETAILED DESCRIPTION OF THE INVENTION
[0020] In one aspect of the present invention there is provided a
composition comprising a protein hydrolysate and a plant sterol,
wherein the weight ratio of the plant sterol to the protein
hydrolysate is from 1:0.02 to 1:150.
[0021] The composition of the present invention can additionally
comprise an emulsifier and/or a fat. Furthermore, the composition
can comprise a mineral salt.
[0022] The weight ratio of the plant sterol to protein hydrolysate
in the compositions is preferably from 1:0.2 to 1:30, more
preferably from 1:0.4 to 1:12.5 and most preferably from 1:1 to
1:5.
[0023] As used here, the term "protein hydrolysate" includes all
hydrolyzed products of proteins prepared by using a proteolytic
enzyme preparation, a microorganism containing suitable proteolytic
activity or acid hydrolysis or any combination thereof, and having
serum lipid profile improving effect. Commercially available
hydrolysates can be used or hydrolysates can be prepared. Preferred
hydrolysates have a molecular weight of 300-100 000 D, more
preferably 500-50 000 D, most preferably 500-30 000 D and/or they
are only slightly soluble in water.
[0024] Plant, animal or microbial proteins and/or their mixtures
can be used as protein sources for the hydrolysates. Proteins of
vegetable origin are preferred. Most preferred are proteins of
grain or legume origin. Suitable vegetable protein sources are for
example soybean protein, wheat protein, especially wheat gluten,
corn protein, oat protein, rye protein, rapeseed or canola protein,
barley protein, rice protein, flaxseed protein, potato protein, pea
protein, lupin protein, maize protein and buckwheat protein. Of
animal origin, suitable examples include milk proteins, such as
whey protein, and their fractions. All these proteins can be used
in different commercially available purified or non-purified forms
as source for the hydrolysates. Also materials containing these
proteins and other major constituents, such as carbohydrates, can
be used as source for the hydrolysates.
[0025] Protein hydrolysates are preferably prepared via enzymatic
hydrolysis by methods and processes well known in the art, but can
also be prepared using other known hydrolytic techniques, such as
acid hydrolysis. For example EP 0 790 060 A1 discloses a method for
preparing suitable protein hydrolysates. Typical preparation
methods include treatment of the protein moiety with a proteolytic
enzyme in aqueous medium, where the protein or protein containing
material is dispersed in water and pH is adjusted with an acid or a
base to the optimum pH range of the enzyme to be used. The enzyme
is preferably added in an amount of 0.2-5%, more preferably 0.5-2%
based on the substrate protein and the reaction is carrier out at
optimum temperature and pH for the time needed depending on the
enzyme and the degree of hydrolysis wanted. The reaction is
typically terminated by heating the mixture to a temperature high
enough to inactivate the enzyme. The reaction mixture can
optionally be neutralized before or after the heating step by using
a suitable acid, e.g. hydrochloric acid, or a base, e.g. sodium
hydroxide. Preferably the fraction having serum lipid profile
improving effect is then separated from the reaction mixture. In
case the water insoluble or slightly water soluble fraction of the
reaction mixture is desired, it can be separated e.g. by
centrifugation or filtration techniques to obtain what is here
referred to as a protein hydrolysate slurry. The protein
hydrolysate slurry can be used as such or as washed with water
and/or dried. The process used for drying is not critical, for
example freeze-drying, spray drying or any other process known in
the art may be used, which produces a powder either directly or
through a grinding step. Dried and powdery protein hydrolysate is
especially preferred as it entraps lipids efficiently. Proteolytic
enzymes that can be used in the preparation of the hydrolysates of
the present invention include enzymes from plant, microbial and
animal origin, including also enzymes from genetically modified
organisms, that at suitable reaction conditions hydrolyse proteins
to hydrolysates that have a serum lipid profile improving effect.
Suitable proteolytic enzymes are e.g. pepsin, trypsin, pancreatin
and papain. It is also possible to use different enzymes
sequentially, e.g. pepsin and trypsin or pepsin and pancreatin.
Especially preferred are the enzymes or combinations of enzymes
that produce protein hydrolysates that are only poorly digestible
or no longer digestible in human intestine and can thus be referred
also as "resistant proteins".
[0026] As used here, the term "plant sterol" refers to any sterol
as defined in the following and having lipid profile improving
effect. Plant sterols include both sterols and saturated sterols
i.e. stanols in either their free form or as esterified e.g. with
fatty acids (2-24 carbon atoms, saturated, monounsaturated or
polyunsaturated, including also special fatty acids such as
conjugated fatty acids, e.g. CLA, and EPA and DHA), hydroxybenzoic
acids or hydroxycinnamic acids (ferrulic or coumaric acids) or
other organic acids such as e.g. di- or tricarboxylic acids and/or
hydroxy acids or with any combination of the said acids. In
addition, the definition of plant sterols also include glycosidic
sterols and their derivatives. Any combinations of the free and
various esterified and glycosylated forms are also included.
[0027] As used here, the term "plant sterol in esterified form" or
"plant sterol ester" refers to plant sterols having at least 60%,
preferably at least 85%, most preferably at least 95% of the plant
sterols in esterified form.
[0028] In this specification the sterols include 4-desmethyl
sterols, 4-monomethyl sterols and 4,4-dimethyl sterols (including
triterpene alcohols) and the stanols include 4-desmethyl stanols,
4-monomethyl stanols and 4,4-dimethyl stanols. Typical 4-desmethyl
sterols are sitosterol, campesterol, stigmasterol, brassicasterol,
22-dehydrobrassicasterol, .DELTA.5-avenasterol. Typical
4,4-dimethyl sterols are cycloartenol, 24-methylenecycloartenol and
cyclobranol. Typical stanols are sitostanol, campestanol and their
24-epimers, cycloartanol and saturated forms obtained by saturation
of e.g. triterpene alcohols (cycloartenol, 24-methylenecycloartenol
and cyclobranol). The definition "plant sterol" includes all
possible natural blends or any mixtures of named sterols and/or
stanols as well as any individual sterol or stanol.
[0029] The amounts of plant sterols in this specification are
calculated as plant sterol equivalents i.e. as the amount of free
plant sterol.
[0030] Commercially available plant sterols in their free or
esterified form can be used as such. When plant sterols in their
free form are used, the particle size of the plant sterols is
preferably reduced to enhance the dispersability, dissolvability
and solubility of the plant sterols. Particle size reduction can be
done by many techniques known in the art, e.g. by different dry or
wet grinding or micromilling techniques described for example in
U.S. Pat. No. 6,129,944, WO 98/58554 and EP 1 142 494 A1. Other
components, such as a suitable admixture can be pulverized together
with plant sterols, the choice of the other components depending on
the food material, nutraceutical or pharmaceutical in which the
active ingredients are to be added. Examples of the admixture
include various structure and flavor enhancers, as well as flours
especially in case the active ingredients are to be added into
bakery products.
[0031] Plant sterols in their free form may also be used molten,
especially in compositions containing an emulsifier and/or a fat.
Preferably a homogenous mixture that is easily used in the
compositions of the present invention is formed from plant sterols
and an emulsifier and/or a fat by heating plant sterols to their
melting point, to 60-150.degree. C., typically to 130-150.degree.
C., and adding the emulsifier and/or fat to the sterols, either
prior to or after heating. Suitable techniques that can be utilized
are described e.g. in U.S. Pat. No. 6,190,720 B1. Most preferably a
blend of plant sterols and emulsifier(s) and/or fat is heated until
the components are dissolved. The mixture is cooled under agitation
prior to adding it into the compositions of the present
invention.
[0032] Preferably the plant sterol is esterified and most
preferably it is a plant sterol fatty acid ester. The fatty acid
ester is technically very suitable for incorporation into a wide
range of different products and is especially preferred as it has
very good organoleptic properties, enabling production of the
compositions of the present invention with good organoleptic
properties.
[0033] Preferably the plant sterol is a stanol because its
absorption is negligible and the use of stanol is therefore safer.
Also the cholesterol lowering effect seems to be stronger with
stanols. Most preferred are therefore the stanol fatty acid esters
for use in the compositions according to the invention.
[0034] As used here, the term "emulsifier" refers to a substance
promoting the formation and improving the stability of emulsions.
Emulsifiers are synthetically produced, derived through chemical
modification of naturally occurring materials or natural products.
The characterizing feature of emulsifiers is a structure in which
one portion of the molecule is polar (hydrophilic) and the other
non-polar (hydro-phobic), which allows the emulsifier to align and
stabilize the contact surfaces of the two immiscible phases of the
emulsion. The relative size and character of the polar and
non-polar portions of the molecule, among other factors, affect the
type and quality of emulsions produced. HLB (hydrophilic lipophilic
balance) scale is commonly used to generally describe the
emulsifying properties of emulsifiers. An emulsifier with a low HLB
value (up to about 6) tends to promote W/O emulsions, an emulsifier
with an intermediate HLB value W/O or O/W emulsions and an
emulsifier with a high HLB value (from about 8) O/W emulsions.
[0035] Preferably the emulsifier or a mixture of emulsifiers is
used to produce a complex with plant sterols and/or protein
hydrolysates. Typical, but non-restricting examples of suitable
emulsifiers include monoglycerides, such as distilled
monoglycerides, diglycerides, monoglyceride derivatives, such as
acetic, lactic, succinic or citric acid esters of monoglycerides,
lecithins, modified lecithins such as enzyme modified lecithin, for
example lysolecithin, polyglycerol esters, polyglycerol
polyricinoleate, sorbitan esters, polysorbates, propylene glycol
esters, stearoyl lactylates, such as sodium stearoyl lactylate and
calcium stearoyl lactylate, diacetyl tartaric acid esters, diacetyl
lactic acid esters, sugar esters, and mixtures thereof. Also lipid
based grain fractions having emulsifying properties can be used as
emulsifiers. These include lipid fractions e.g. from oat.
[0036] Preferred emulsifiers or mixtures of emulsifiers are those
having a HLB value or emulsifying properties equivalent to a HLB
value above 4, more preferably above 6. Examples of preferred
emulsifiers include acetic, succinic, lactic and citric acid esters
of monoglycerides, lecithins, modified lecithins, polyglycerol
esters, polysorbates, diacetyl tartaric acid esters,
monoglycerides, stearoyl lactylates and mixtures thereof.
[0037] As used here, by "synthetic emulsifier" is meant emulsifiers
that are synthetically produced or derived through chemical
modification of naturally occurring materials. These synthetic
emulsifiers do not have strong color, unlike some naturally
occurring emulsifiers, such as lecithin and enzymatically modified
lecithin. The synthetic emulsifiers can thus be used as a part of
protein hydrolysate/emulsifier complexes added also into light
colored products. The synthetic emulsifier are also often more pure
than the naturally occuring emulsifiers. Suitable, but non-limiting
examples of the synthetic emulsifiers are monoglyceride
derivatives, polyglycerol esters, propylene glycol esters,
polysorbates, sorbitan esters, stearoyl lactulates, diacetyl
tartaric acid esters, sugar esters and mixtures thereof Preferred
synthetic emulsifiers or mixtures of synthetic emulsifiers are
those having a HLB value or emulsifying properties equivalent to a
HLB value above 4, more preferably above 6.
[0038] As used here, the term "complex" is a composition in which
only a part of the emulsifier(s) and/or fat and/or plant sterols
are extractable by using non-polar solvents commonly used to
extract "free lipids". These non-polar solvents include e.g.
petroleum ether and hexane. The rest of the emulsifier(s) and/or
fats and/or plant sterols can only be extracted after hydrolysis
e.g. by using methods including acid hydrolysis, such as AOAC
922.06.
[0039] As used here, by "fat" is meant edible solid fats,
semi-solid fats, liquid oils and any mixtures thereof Also fat
substitutes such as sugar polyesters are included. Examples of
solid and semi-solid fats that are usable include tallow, lard,
butter, margarine and shortenings as well as semi-solid tropical
oils, such as coconut oil and palm kernel oil. Typical examples of
liquid oils include canola/rapeseed oil, soybean oil, sunflower
oil, olive oil, palm oil, corn oil, sesameseed oil, cottonseed oil,
wheat germ oil, peanut oil and fish oils. Liquid vegetable oils are
especially preferred. The fat can be naturally occurring or
modified, for example hydro-genated, transesterified, contain
structured triacylglycerols or an increased amount of
diacylglycerols.
[0040] As used here, the term "mineral salt" includes nutritionally
beneficial mineral salts having elevated K and/or Mg and/or Ca
content and/or reduced Na content compared to common salt. In U.S.
Pat. No. 6,136,349 this kind of mineral salt, especially when
administered together with plant sterols and/or used to replace
common salt in food items, is suggested to bring beneficial health
effects, such as lowering elevated blood pressure. Examples of
suitable mineral salt compositions include mineral salt sold by
trade name Pansalt.RTM. or Cardia.RTM. salt. The mineral salt is
preferably added into the protein hydrolysate, more preferably into
the protein hydrolysate slurry or washed protein hydrolysate.
[0041] In the following some typical compositions of the present
invention and the preparation thereof will be explained in more
detail.
[0042] The composition of the present invention comprising one or
more protein hydrolysates and one or more plant sterols can be used
as such or as a pharmaceutical or a nutraceutical or most
advantageously in food products for improving serum lipid profile
in man. The protein hydrolysate(s) and plant sterol(s) may be added
in any suitable way into food products, pharmaceuticals or
nutraceuticals separately as ingredients or they may be suitably
combined to form compositions or complexes before forming the
composition according to the invention. Preparation of the
compositions and the incorporation into food products,
pharmaceuticals or nutraceuticals can be facilitated by reducing
particle size of said compositions or components thereof e.g. by
grinding or micromilling. Also other components such as water or
mineral salt may be added. Typical but non-restricting examples of
the preparation of the compositions and complexes are described in
the following embodiments.
[0043] In one embodiment of the present invention the protein
hydrolysate(s) and the plant sterol(s) are added into food
products, pharmaceuticals or nutraceuticals separately as
ingredients. The active ingredients are added into food material,
pharmaceuticals or nutraceuticals by conventional processes for
producing these products.
[0044] Commercially available plant sterols in their free form can
be used as such, or more preferably the particle size of the plant
sterols is reduced to enhance the dispersability and solubility of
plant sterols as described above. Molten plant sterols in their
free form can also be used. Advantageously the main part of plant
sterols is in esterified form and preferably melted, if needed,
before addition into food products, pharmaceuticals or
nutraceuticals. Protein hydrolysates can be used in any suitable
form including protein hydrolysate slurry, washed and/or dried
protein hydrolysate, protein hydrolysate in an aqueous solution or
protein hydrolysate with reduced particle size.
[0045] In another embodiment of the present invention the protein
hydrolysate(s) and the plant sterol(s) are combined into a
composition that can be used as such or added into pharmaceuticals
or nutraceuticals or food products. When plant sterols in their
free form are used, they are mixed with dried protein hydrolysates
to form a homogenous composition or more preferably homogenized
with the protein hydrolysate slurry or water washed protein
hydrolysate to form a protein hydro-lysate/plant sterol complex.
The mixture may be used as such or preferably be dried by methods
commonly used in the art, e.g. by freeze drying or spray drying, to
form powder either directly or through a grinding step.
Commercially available plant sterols can be used as such or
preferably with reduced particle size. In a preferred embodiment
the main part of the plant sterols is in esterified form.
Esterified plant sterols are preferably mixed with a dried and
powdery protein hydrolysate to form a protein hydrolysate/plant
sterol complex.
[0046] In still another embodiment of the present invention the
protein hydrolysate(s), mineral salt and the plant sterol(s) are
combined into a composition that can be used as such or added into
pharmaceuticals or nutraceuticals or food products. The mineral
salt is preferably added into the protein hydrolysate, more
preferably into the protein hydrolysate slurry or washed protein
hydrolysate and the mixture is combined with plant sterols.
[0047] Alternatively the compositions comprising the protein
hydrolysate(s) and the plant sterol(s) can be prepared by exposing
a mixture of unhydrolyzed protein(s) and plant sterol(s) to
hydrolysis, preferably to enzymatic hydrolysis by methods and
processes known in the art.
[0048] The composition of the present invention comprising one or
more protein hydro-lysates, one or more emulsifiers and/or fats and
one or more plant sterols can be used as such or as pharmaceuticals
or nutraceuticals or most advantageously in food products for
improving serum lipid profile in man. The protein hydrolysate(s),
plant sterol(s) and emulsifier(s) and/or fat(s) may be added in any
suitable way into food products, pharmaceuticals or nutraceuticals
separately as ingredients or they may be suitably combined to form
compositions or complexes before forming the composition according
to the invention, which is incorporated in the food products,
pharmaceuticals or nutraceuticals. Preparation of the compositions
and the incorporation into food products, pharmaceuticals or
nutraceuticals can be facilitated by reducing particle size of said
compositions or components thereof e.g. by grinding or
micromilling. Also other components such as water or mineral salt
may be added. Typical but non-restricting examples of the
preparation of the compositions and complexes are described in the
following embodiments. Preferably the emulsifier is used to produce
a complex with protein hydrolysate(s) and/or plant sterol(s).
[0049] In one embodiment of the present invention the protein
hydrolysate(s) and the emulsifier(s) are used to form a protein
hydrolysate/emulsifier complex, and the complex and the plant
sterols are added into food products, pharmaceuticals or
nutraceuticals separately as ingredients. The active ingredients
are added into food material, pharmaceuticals or nutraceuticals by
conventional processes for producing these products.
[0050] The protein hydrolysate/emulsifier complex can be prepared
by mixing the emulsifier and protein hydrolysate e.g. by a high
speed mixer, or more preferably by forming a protein
hydrolysate/emulsifier complex by mixing the ingredients in the
presence of water. Preferably the emulsifier is dispersed in water,
protein hydro-lysate added, followed by thorough mixing at room
temperature. Optionally the complex is dried by conventional drying
techniques, such as e.g. freeze-drying or spray-drying, to form a
powdery product either directly or through a grinding step. The
complex can also be prepared of the protein hydrolysate and a
mixture of emulsifier and fat, with or without the presence of
water. The protein hydrolysate/emulsifier complex contains
preferably at least 5%, more preferably at least 10%, and most
preferably at least 20% emulsifier on dry weight basis Also any
commercially available protein hydrolysate/phospholipid complex can
be used as protein hydrolysate/emulsifier complex.
[0051] Prior art discloses protein/phospholipid and protein
hydrolysate/phospholipid complexes containing 10% or more bound
phospholipid (EP 0 790 060 A1), especially lecithin or enzyme
modified lecithin, where both the protein or protein hydrolysate
and phospholipids have lipid metabolism improving effect. In the
present invention a wider range of emulsifiers or their
combinations can be used. This is of particular benefit when the
complex is added into food products, pharmaceuticals or
nutraceuticals, where the emulsifiers can be selected to bring
advantageous effects on the properties, e.g. texture or taste, of
the final product and not just solely on the basis of enhancing the
lipid profile improving effect of the protein hydrolysate.
[0052] Commercially available plant sterols in their free form can
be used as such, or more preferably the particle size of the plant
sterols is reduced to enhance the dispersability and solubility of
plant sterols. Particle size reduction can be done by many
techniques known in the art, e.g. by grinding. Advantageously the
main part of plant sterols is in esterified form and preferably
melted, if needed, before addition into food products,
pharmaceuticals or nutraceuticals.
[0053] In another embodiment of the present invention plant sterols
in a form of a plant sterol/emulsifier complex, or plant sterols
dissolved or suspended in a fat, and protein hydrolysate are added
into food products, pharmaceuticals or nutraceuticals separately as
ingredients. The active ingredients are added into food material,
pharmaceuticals or nutraceuticals by conventional processes for
producing these products. The plant sterol/emulsifier complex or
the suspension or solution containing plant sterols and fat is
prepared by dissolving or suspending plant sterols in their free
form, preferably with reduced particle size, in a melt of an
emulsifier, a fat or a mixture of emulsifier and fat at elevated
temperatures, preferably at >60.degree. C., more preferably at
>80.degree. C., mixing to form a homogenous dispersion or
solution and cooling with agitation. Also molten plant sterols may
be used. The weight ratio of plant sterols to emulsifier can be
from 1:0.01 to 1:5, preferably from 1:0.05 to 1:2, more preferably
from 1:0.1 to 1:2. The crude fat content, excluding plant sterols
and emulsifiers, is preferably less than 80%, more preferably less
than 60%, most preferably less than 20% of the weight of the
complex or dispersion or solution. Also plant sterols mainly in
their esterified form may be used instead of plant sterols in their
free form. Protein hydrolysates can be used in any suitable form
including protein hydrolysate slurry, washed and/or dried protein
hydrolysate, protein hydrolysate in an aqueous solution, protein
hydrolysate with reduced particle size and protein
hydrolysate/emulsifier complex.
[0054] In a preferred embodiment of the present invention the plant
sterol(s), the emulsifier(s) and the protein hydrolysate(s) are
combined into a protein hydro-lysate/plant sterol/emulsifier
complex that can be used as such or added into pharmaceuticals or
nutraceuticals or food products.
[0055] The complex is preferably prepared by first dissolving or
suspending plant sterols in a melt of an emulsifier, a fat or a
mixture of emulsifier and fat at elevated temperatures, preferably
at >60.degree. C., more preferably at >80.degree. C., mixed
to form a homogenous dispersion or solution and cooled with
agitation. The weight ratio of plant sterols to emulsifier can be
from 1:0.01 to 1:5, preferably from 1:0.05 to 1:2, more preferably
from 1:0.1 to 1:2. The fat content, excluding plant sterols and
emulsifiers, is preferably 0-80%, more preferably 0-60%, most
preferably 0-20% of the weight of the complex or suspension or
solution. Protein hydrolysate in a dry form, as a slurry or a water
washed protein hydrolysate or an aqueous protein hydrolysate
suspension or solution is then mixed and homogenized with the
cooled plant sterol/emulsifier or plant sterol/fat or plant
sterol/emulsifier/fat mixture to form a protein hydrolysate/plant
sterol/emulsifier complex or if no emulsifier is present, a protein
hydrolysate/plant sterol complex. The homogenized mixture can
optionally be dried and/or its particle size reduced before usage.
Plant sterols in their free form are preferably used. Plant sterols
in their free form with reduced particle size are especially
preferred. Also molten plant sterols may be used. Plant sterols
mainly in their esterified form may also be used. The complex may
also be prepared by combining a protein hydrolysate/emulsifier
complex and plant sterols.
[0056] Alternatively the compositions consisting the protein
hydrolysate(s), the emulsifier(s) and/or fat(s) and the plant
sterol(s) can be prepared by first exposing a complex of
unhydrolyzed protein/emulsifier or unhydrolyzed protein/plant
sterols or unhydrolyzed protein/plant sterols/emulsifier(s) to
hydrolysis, preferably to enzymatic hydrolysis by methods and
processes known in the art and described earlier, and then
combining the hydrolyzed complex with other components according to
the present invention.
[0057] In a second aspect of the present invention there is
provided a composition comprising a protein hydrolysate and
emulsifier(s) or emulsifier-fat mixture for improving serum lipid
profile in man, wherein the synthetic emulsifier(s) are used. By
using synthetic emulsifiers, the taste, colour and impurity
problems commonly attributed to compositions containing natural
emulsifiers can be avoided. Also the lipid based grain fraction(s)
having emulsifying properties can be used as emulsifiers in a
composition comprising a protein hydrolysate and emulsifier(s) or
emulsifier-fat mixture. In this third aspect of the invention the
protein hydrolysate and the emulsifier are preferably obtained from
the same source. E.g. oat is fractionated and a protein rich
fraction is hydrolysed and combined with a lipid rich fraction to a
mixture or a complex. A benefit with this combination is that it is
made of non-allergenic materials.
[0058] The composition of the present invention comprising one or
more protein hydro-lysates and one or more synthetic emulsifiers
and optionally fat(s), and/or one or more lipid based grain
fractions having emulsifying properties, can be used as such or as
pharmaceuticals or nutraceuticals or most advantageously in food
products for improving serum lipid profile in man. The protein
hydrolysate(s) and synthetic emulsifier(s), and optionally fat(s),
or the protein hydrolysate(s) and lipid based grain fractions
having emulsifying properties may be added in any suitable way into
food products, pharmaceuticals or nutraceuticals separately as
ingredients or they may be suitably combined to form compositions
or complexes before forming the composition according to the
invention, which is incorporated in the food products,
pharmaceuticals or nutraceuticals. Preferably the synthetic
emulsifier or lipid based grain fraction is used to produce a
complex with protein hydrolysate(s) by methods described
earlier.
[0059] In a further aspect of the present invention there is
provided a food product comprising at least one basic nutritional
ingredient and at least one of the compositions of the present
invention as defined above.
[0060] The plant sterol constituent of the composition is present
in a higher amount than naturally occurring in the basic
nutritional ingredient(s).
[0061] The food product of the present invention can be prepared by
adding said composition to food material(s) by the conventional
processes for producing food products. The composition can be added
as such (either in liquid, semi-solid or dried form) or the
constituents of the composition can be added separately as
ingredients.
[0062] The food product of the present invention can be in the form
of various food compositions, including but not restricted to
[0063] bakery products and confectionery (fresh and dry bakery
products, e.g. fresh bread, other bread products, cakes, muffins,
waffles, biscuits, crackers, protein enriched bakery products etc.)
[0064] cereal products and snacks (breakfast cereals, muesli, bars,
such as cereal based and muesli bars, such bars possibly containing
chocolate, pasta products, flours etc.) [0065] beverages (alcoholic
and non-alcoholic drinks, including e.g. soft drinks, juices and
juice-type mixed drinks, fortified beverages such as e.g. protein
or calcium fortified beverages, probiotic drinks, sports and energy
drinks, dietary supplement and meal replacement drinks,
concentrates or premixes for beverages and powdered drinks where
the content of compositions of the present invention is calculated
for the ready-to-use form, etc.) [0066] dairy products (milk, milk
based products, e.g. cheese, cream cheese and the like, yogurt,
frozen yogurt, other frozen dairy foods, drinkable yogurt, other
fermented milk products, dairy beverages, ice cream, desserts,
spreads etc.) [0067] fermented cereal products [0068] sauces, soups
[0069] meat, fish, poultry and egg products (e.g. sausages, meat
balls etc.) [0070] analogues for dairy or meat products (e.g.
imitations of cheese, yogurt, ice cream, desserts, meat
substitutes, milk alternatives etc.), non-dairy frozen desserts
[0071] soy based products [0072] vegetable oil based products (e.g.
margarines, spreads, dressings, mayonnaise etc.) [0073] ready mixes
(for baking e.g. breads, cakes, muffins, waffles, pizzas, pancakes;
or for cooking e.g. soups, sauces, desserts, puddings) to be used
in preparing or manufacturing foods.
[0074] The food product of the present invention can also contain
other nutritionally beneficial components, some of which may
further enhance the effects of the compositions of the present
invention. The food can be fortified with these components or the
components can be an intrinsic part of the other food
ingredients.
[0075] Examples of the nutritionally beneficial components include
diacylglycerol and n-3 fatty acids, e.g. from fish oil, which have
advantageous effects on the lipid metabolism and may thus reduce
the risk of cardiovascular disease. N-3 fatty acids act favourably
on blood characteristics, e.g. they are hypotriglyceridemic and
reduce platelet aggregation. Diacylglycerol reduces serum
triacylglycerol levels and can thus have a favourable effect on the
risk of cardiovascular disease. Both n-3 fatty acids and
diacylglycerols may lower risk of atherosclerosis and CVD mortality
also by other mechanisms.
[0076] Other non-limiting examples of the beneficial nutritional
components include dietary fibre and beneficial minor components,
for example such as isoflavones, tocopherols, tocotrienols,
carotenoids, vitamin C, folate and flavonoids. Also other vitamins
and minerals may be added or included in the food products of the
present invention.
[0077] Especially the use of dietary fibre may further enhance the
effects of the present compositions. By dietary fibre, it is meant
food components derived from plant material, or analogous
carbohydrates from other sources, that are resistant to digestion
and absorption by human digestive enzymes. This includes various
polysaccharides, oligosaccharides, lignins and associated
substances that are resistant to digestion. Dietary fibre may
further be classified into water-soluble and water insoluble
fractions. Examples of the water-soluble fraction are e.g. pectin,
plant gums, .beta.-glucans and resistant starch and of the
insoluble fraction e.g. cellulose and hemicellulose. An adequate
fibre intake is thought to have many beneficial effects, for
example viscous soluble fibre is known to reduce risk of
cardiovascular disease by lowering plasma cholesterol levels. In a
further aspect of the present invention there is provided a
pharmaceutical or nutraceutical product for improving serum lipid
profile comprising a composition of the present invention as
defined above. Said product can additionally contain at least one
compounding agent. The compounding agent can be any
pharmaceutically acceptable binder, carrier, diluent, excipient or
coating agent. The product can be in any suitable form, e.g.
tablets, granules, powder, capsules, syrups, dispersions or other
liquid preparations for oral administration.
[0078] The products and compositions of the present invention
preferably contain at least two active ingredients, i.e. protein
hydrolysate(s) and plant sterol(s), in sufficient amounts to
provide improvement in serum lipid profile, e.g. reduction in serum
total and/or LDL cholesterol levels. The invention is especially
directed to synergistic combination of at least two active
ingredients, i.e. protein hydrolysate(s) and plant sterol(s), the
use of which leads to enhanced improvements in serum lipid profile
compared to the effects obtained when one of the ingredients is
used alone. Certain protein hydrolysate/plant sterol/emulsifier
complexes according to the present invention are especially
beneficial as the lipid profile improving effect is optimized by
ensuring optimal availability of the active ingredients at the
critical sites of action. In addition to reduction of serum total
and LDL cholesterol levels, some compositions of the present
invention also provide desired increase in serum HDL cholesterol
levels. Some of the compositions also have a beneficial effect on
blood pressure. The invention is also directed to combinations of
active ingredient(s) and ingredient(s) ensuring the optimal
availability of the active ingredient at the site of action, i.e.
protein hydrolysate(s) and synthetic emulsifier(s).
[0079] The present invention also makes it possible to reduce the
daily intake of plant sterols in man and still achieving similar
cholesterol lowering effects as obtained with the generally
recommended daily intake of plant sterols (2-3 g) which can be
achieved e.g. by consuming commercial products enriched with plant
sterols. The optimal daily intake of the compositions of the
present invention as such or when used in foods, nutraceuticals or
pharmaceuticals is such that a daily intake of plant sterols
(calculated as sterol equivalents) of 0.4-5 g, preferably 0.5-2.5
g, more preferably 0.8-2 g is supplied.
[0080] According to the present invention it is also possible to
lower the protein hydro-lysate content of food products containing
protein hydrolysates especially purposed for improving serum lipid
profile, thus improving the organoleptic properties of such foods.
The optimal daily intake of the compositions of the present
invention as such or when used in foods, nutraceuticals or
pharmaceuticals is such that a daily intake of protein hydrolysate
of 0.1-60 g, preferably 0.5-15 g, more preferably 0.8-10 g is
supplied.
[0081] The invention is also directed to food products containing
at least 10% of the optimal daily intake of the plant sterols and
protein hydrolysates per serving. Preferably the food products
contain from 10% to 200% of the optimal daily intake of plant
sterols and protein hydrolysates per serving.
[0082] Typically the plant sterol content of the food products is
between 0.05-20 g per 100 g food product, preferably 0.1-15 g/100
g, more preferably 0.1-10 g/100 g, and most preferably 0.5-5 g/100
g and the protein hydrolysate content is between 0.0002-20 g per
100 g food product, preferably 0.0005-15 g/100 g, more preferably
0.001-10 g/100 g, and most preferably 0.1-10 g/100 g.
[0083] In yet a further aspect of the present invention there is
provided a method for improving serum lipid profile e.g. for
lowering total and/or LDL cholesterol, increasing HDL cholesterol,
reducing triacylglycerol, reducing apolipoprotein B and/or
increasing the ratio of HDL cholesterol to LDL cholesterol in a
subject, comprising orally administering to the subject a
composition according to the present invention in an amount
effective for improving serum lipid profile.
[0084] Further, the present invention provides a method to reduce
or prevent the development of atherosclerosis in humans by dietary
means including orally administering to the subject a composition
according to the present invention in an amount effective for
improving serum lipid profile.
[0085] The main advantages of the present invention obtained by
combining the intake of the two active ingredients are: [0086]
synergistic serum lipid profile improving effect of plant sterols
and protein hydrolysates [0087] enhanced serum lipid profile
improving effect compared to what is achievable by using plant
sterols or protein hydrolysates alone [0088] lower daily intake of
plant sterols is needed to achieve the same serum lipid profile
improving effect than what is achieved by using plant sterols alone
[0089] lower daily intake of protein hydrolysates is needed to
achieve the same serum lipid profile improving effect than what is
achieved by using protein hydro-lysates alone [0090] lowering the
risk of cardiovascular disease by means of improving serum lipid
profile.
[0091] The present invention is especially directed to enhanced
serum lipid profile improving effect by utilizing the synergistic
combination of plant sterols and protein hydrolysates, optionally
with emulsifiers and/or fat and/or mineral salt. The present
invention is also directed to ensuring optimal availability of the
active ingredients, especially protein hydrolysates, at the site of
action. Certain compositions containing protein hydrolysates and
emulsifiers or protein hydrolysates, emulsifiers and plant sterols
according to the current invention are especially beneficial as the
lipid profile improving effect is optimized by ensuring optimal
availability of the active ingredients.
[0092] The present invention is also especially directed to an
optimized daily intake of plant sterol and protein hydrolysate to
achieve a significant serum lipid profile improving effect in a
form of a food product as part of the daily diet. The daily intake
of plant sterols can be decreased by combining the beneficial
effect of plant sterols and other lipid profile improving dietary
components to achieve similar lipid profile improving effect as
obtained with currently recommended daily intake (2-3 g/d) of plant
sterols. This approach advantageously reduces the daily intake of
plant sterols, leading to a more balanced use of plant sterols. The
synergistic effect is especially profound at daily intakes of at
most 2 g plant sterol.
[0093] In a preferred embodiment of the invention the compositions
contain plant sterols and soy protein hydrolysate, optionally with
emulsifier(s). Preferably the plant sterol used is a plant sterol
fatty acid ester and even more preferred it is a plant sterol fatty
acid ester containing a substantial amount of stanol fatty acid
ester, preferably at least 30% stanol fatty acid ester.
[0094] The compositions according to the present invention are most
preferably incorporated into foods designed for being a part of a
healthy diet.
[0095] Additional aspects of the invention are:
[0096] Use of a composition according to any one of claims 1 to 13
for the preparation of a pharmaceutical, nutraceutical or food
product for improving serum lipid profile, especially for lowering
serum total and/or LDL cholesterol and/or for increasing the ratio
of HDL cholesterol to LDL cholesterol and/or for lowering the serum
apolipoprotein B level.
[0097] Use of a composition according to any one of claims 1 to 13
for the preparation of a pharmaceutical, nutraceutical or food
product for reducing or preventing the development of
atherosclerosis.
[0098] The above uses wherein the protein hydrolysate is
administrated at a rate of 0.1 to 60 g, preferably 0.5 to 15 g and
more preferably 0.8 to 10 g per day, and the plant sterol is
administrated at a rate of 0.4 to 5 g, preferably 0.5 to 2.5 g and
more preferably 0.8 to 2 g per day calculated as sterol
equivalents.
[0099] A method for improving serum lipid profile, especially for
lowering serum total and/or LDL cholesterol and/or for increasing
the ratio of HDL cholesterol to LDL cholesterol and/or for lowering
the serum apolipoprotein B level in a subject, comprising orally
administering to the subject a composition according to any one of
claims 1 to 13, in an amount effective for improving serum lipid
profile.
[0100] A method for reducing or preventing the development of
atherosclerosis in a subject by dietary means including orally
administering to the subject a composition according to any one of
claims 1 to 13, in an amount effective for improving serum lipid
profile.
[0101] The above methods wherein the protein hydrolysate is
administrated at a rate of 0.1 to 60 g, preferably 0.5 to 15 g and
more preferably 0.8 to 10 g per day, and the plant sterol is
administrated at a rate of 0.4 to 5 g, preferably 0.5 to 2.5 g and
more preferably 0.8 to 2 g per day calculated as sterol
equivalents.
[0102] The invention is further illustrated by the following
examples. Examples 1-4 illustrate preparation of the constituents
of the compositions, examples 5-8 illustrate preparation of the
compositions according to the invention, examples 9-16 illustrate
the use of the compositions in food products and example 17
illustrates the effect of a composition of the present invention in
lowering serum cholesterol. In this specification the percentages
mean % by weight unless otherwise specified.
Example 1
[0103] Preparation of Protein Hydrolysate.
[0104] 1500 g isolated soy protein (SUPRO.RTM. Brand Isolated Soy
Protein, Protein Technologies International) was dispersed in 15 l
of water, pH was adjusted to 2 with HCl, and pepsin (P7000, Sigma
Aldrich, activity 1:10,000) was added to the solution (1% of the
amount of isolated soy protein). The reaction was carried out at
37.degree. C., for 24 hours and stopped by heating the reaction
mixture to 85.degree. C. for one hour. The mixture was neutralized
with sodium hydroxide (2 mol/l) and centrifuged. The precipitate
was washed twice with water after which the protein hydrolysate was
freeze-dried and ground. 300 g dried protein hydrolysate was
obtained. The molecular weight of the peptides obtained in the
hydrolysis was determined by SDS-polyacrylamide gel electrophoresis
(SDS-page) method. The molecular weight range of the peptides in
the hydrolysate was from 3000 to 30000 D.
Example 2
[0105] Preparation of Protein Hydrolysate/Emulsifier Complex.
[0106] 62 g of emulsifier (lysolecithin, Precept 8160, Central
Soya) was dispersed in water (2500 ml) in room temperature and 500
g soy protein hydrolysate prepared as in example 1 was added. The
solution was mixed by a high speed mixer (10 000 rpm), the mixture
was freeze-dried and ground to obtain protein
hydrolysate/emulsifier complex.
[0107] The total lipid content of the complex was determined by an
acid hydrolysis method (AOAC 922.06). The lipids that were not
bound or only loosely bound to the complex were determined by
direct petroleum ether extraction (free lipids). The content of
bound lipids was calculated as the difference of total and "free"
lipids. Total lipid content was 11.0% of the complex, free lipid
content 7.9% of the complex and thus the "bound" lipid content was
3.1% of the complex.
Example 3
[0108] Preparation of Plant Sterol/Emulsifier Complex Using Plant
Sterols in Their Free Form.
[0109] Plant sterol/emulsifier complex was prepared of plant
sterols in their free form (Phytosterols, Archer Daniels Midland
Company), sodium stearoyl lactylate (Grindsted SSL P 55, Danisco
Cultor) and citric acid ester of monoglycerides (Grindsted.TM.
Citrem P 70, Danisco Cultor). Sodium stearoyl lactylate (30 g,
beads) and citric acid esters of monoglycerides (70 g, paste) were
melted at 60.degree. C. and mixed homogenous. Plant sterols (100 g)
were added into the emulsifier mixture, the ingredients were mixed
together and heated under agitation to about 145.degree. C. Clear
liquid obtained was then cooled with stirring to 70.degree. C. and
used for preparation of a salad dressing (in the example 11).
Example 4
[0110] Preparation of Plant Sterol/Emulsifier Complex Using Plant
Sterols Both in Their Free and Esterified Forms.
[0111] Plant sterol/emulsifier complex was prepared of a mixture of
plant sterols in their free and esterified forms and lecithin.
Plant sterol ester (150 g, Sterol ester-115, Raisio Benecol) was
molten at 80.degree. C. Plant sterols (100 g, Phytosterols, Archer
Daniels Midland Company), with reduced particle size, and lecithin
(100 g, Adlec, Archer Daniels Midland Company) were mixed into
molten sterol ester and the mixture was heated under agitation to
melt the crystalline plant sterols. The product was homogenized
with a high speed mixer (25 000 rpm) and cooled with stirring to
60.degree. C.
Example 5
[0112] Preparation of Protein Hydrolysate/Emulsifier Complex by
Using a Synthetic Emulsifier.
[0113] 35 g of emulsifier (citric acid ester of monoglycerides,
Grindsted.TM. Citrem P 70, Danisco Cultor) was dispersed in water
(1500 ml) at 50.degree. C. and 150 g of the soy protein hydrolysate
prepared in example 1 was added. The solution was mixed by a high
speed mixer (25 000 rpm), the mixture was freeze-dried and ground
to obtain protein hydrolysate/emulsifier complex.
Example 6
[0114] Preparation of Protein Hydrolysate/Plant Sterol Complex
Using Plant Sterols in Their Free Form.
[0115] Isolated soy protein (500 g, SUPRO.RTM. Brand Isolated Soy
Protein, Protein Technologies International) was hydrolyzed as in
example 1 and washed twice with water after hydrolyzation. Plant
sterols in their free form (40 g, Phytosterols, Archer Daniels
Midland Company) were added and homogenized with the washed protein
hydrolysate with a high speed mixer (25 000 rpm). The composition
was freeze-dried and ground.
Example 7
[0116] Preparation of Protein Hydrolysate/Plant Sterol Complex
Containing Mineral Salt, Using Plant Sterols in Their Esterified
Form.
[0117] Isolated soy protein (500 g, SUPRO.RTM. Brand Isolated Soy
Protein, Protein Technologies International) was hydrolyzed as in
example 1 and washed twice with water after hydrolyzation. Mineral
salt (5 g, Pansalt.RTM.) was added to the washed hydrolysate slurry
and the slurry was freeze-dried and ground. 34.4 g plant sterol
ester (Sterol ester-115, Raisio Benecol) was added to 65.6 g of the
dried hydro-lysate. The mixture was homogenized with a high speed
mixer (25 000 rpm).
Example 8
[0118] Preparation of Protein Hydrolysate/Plant Sterol/Emulsifier
Complex.
[0119] Soy protein hydrolysate was prepared as in example 1. Enzyme
modified lecithin (20 g, Precept 8160, Central Soy Company) and
citric acid esters of monoglycerides (50 g Grindsted.TM. Citrem P
70, Danisco Cultor) were melted at 60.degree. C. and mixed
homogenous. Plant sterols (50 g, Phytosterols, Archer Daniels
Midland Company) with reduced particle size were added into the
emulsifier mixture, the ingredients were mixed together and heated
under agitation to 100.degree. C. Clear liquid obtained was then
cooled with stirring until the temperature reached 90.degree. C.
Dried and ground soy protein hydrolysate (90 g) was added and
homogenized into the plant sterol emulsifier blend at 90.degree. C.
The mixture was cooled to 70.degree. C. and used for preparation of
a milk alternative (in the example 15) and a fruit drink (in the
example 16).
[0120] The weight ratio of plant sterol to protein hydrolysate in
the complex was 1:1.8 and the complex contained 23.8% plant sterols
and 42.9% soy protein hydrolysate.
Example 9
[0121] Food Bar with Protein Hydrolysate/Plant Sterol Complex.
[0122] Food bar was prepared of the following ingredients:
TABLE-US-00001 300 g oat flakes 95 g fiber rich oat bran 140 g corn
syrup 50 g brown sugar 40 g concentrated apple juice 20 g apple 30
g raisin 100 g vegetable fat 3 g salt 8 g flavoring 110 g complex
from example 6 4 g lecithin A 45 g food bar contained 1.6 g plant
sterols and 3.9 g soy protein hydrolysate.
Example 10
[0123] Yogurt with Protein Hydrolysate/Emulsifier Complex and Plant
Sterols.
[0124] Plant stanol ester (STAEST-115, Raisio Benecol) and protein
hydrolysate/emulsifier complex from example 5 were used separately
as ingredients in preparation of yogurt. Plant stanol ester
(STAEST-115) was molten at 70.degree. C. Molten stanol ester (110
g) and 140 g soy protein hydrolysate/emulsifier complex from
example 5 were added into 5.75 l of skimmed milk at 70.degree. C.,
mixed with a high speed mixer and the mixture was pasteurized.
Normal yogurt cultures and Bifidobacteria were stirred into the
mixture and the mixture was held at 42.degree. C. for 7 hours.
[0125] A 150 g serving of the product contained 1.7 g plant stanols
and 3.0 g protein hydrolysate.
Example 11
[0126] Low-fat salad dressing with plant sterol/emulsifier complex
and protein hydro-lysate.
[0127] Plant sterol/emulsifier complex from example 3 and soy
protein hydrolysate from example 1 were used separately as
ingredients in a preparation of low-fat salad dressing.
[0128] Ingredients of the salad dressing were:
TABLE-US-00002 7.5 g plant sterol/emulsifier complex from example 3
7 g soy protein hydrolysate from example 1 16 g soybean oil 56 g
water 6 g vinegar 4.5 g sugar 1.5 g salt 0.1 g xanthan gum 0.4 g
lemon juice 1 g spices 30 g serving of the salad dressing contained
1.1 g plant sterols and 2.1 g soy protein hydrolysate.
Example 12
[0129] Yogurt-Like Cereal Product with Plant Sterol/Emulsifier
Complex and Protein Hydrolysate.
[0130] Protein hydrolysate was prepared from wheat gluten (Raisio)
in a similar way as soy protein hydrolysate in example 1, except
the hydrolysate was not dried. Water was added into the washed
gluten hydrolysate to obtain a product having a solid content of
20%.
[0131] Plant sterol/emulsifier complex from example 4 and wheat
gluten hydrolysate were used in a preparation of fermented,
yogurt-like cereal product.
Ingredients
TABLE-US-00003 [0132] 45% water 16.5% prepared mixture of gluten
hydrolysate and water 8.9% oat bran 28% berry jam (containing
fructose, blueberry, strawberry, raspberry, pectin, flavors) 1.6%
plant sterol/emulsifier complex from example 4
[0133] The mixture of water and oat bran was fermented using
Bifidobacteria culture. The berry jam, sterol/emulsifier complex
and mixture of gluten hydrolysate and water were added and all
ingredients worked together.
[0134] A 150 g serving of the product contained 1.3 g plant sterols
and 5.0 g protein hydrolysate.
Example 13
[0135] Bread Rolls with Protein Hydrolysate/Plant Sterol Complex
Containing Mineral Salt.
[0136] Bread rolls containing the complex from example 7 were
prepared in a conventional way of the following ingredients:
TABLE-US-00004 800 g wheat flour 20 g sugar 20 g salt 10 g
margarine (80% fat content) 94 g complex from example 7 500 g water
11 g dried yeast 24 bread rolls were obtained from the dough. Two
50 g bread rolls contained suitable daily doses of plant sterols
(1.3 g) and protein hydrolysate (4.0 g).
Example 14
[0137] Cream Cheese Style Spread with Plant Sterols and Protein
Hydrolysate with Mineral Salt.
[0138] Plant stanol ester (STAEST-115, Raisio Benecol) and soy
protein hydrolysate with mineral salt were used separately as
ingredients in a preparation of cream cheese style spread. Soy
protein hydrolysate containing mineral salt was prepared in a
similar way as the complex in example 7, except no plant sterol
ester was added into the dried and ground protein hydrolysate. The
dried soy protein hydrolysate contained 4.8% mineral salt.
[0139] Plant stanol ester was first incorporated into a fat blend,
composition of which was 59.7% rapeseed oil, 7% interesterified
blend of palm stearine and coconut oil and 33.3% plant stanol ester
(STAEST-115, Raisio Benecol). The blend was prepared by blending
the melted stanol ester with rapeseed oil and the hardstock
component.
[0140] The cream cheese style spread was produced according to the
following recipe:
TABLE-US-00005 51.3% curd 24.6% fat blend including the stanol
ester 12.0% condensate 1.0% stabilizer 1.0% milk proteins 7.9% soy
protein hydrolysate containing 4.8% mineral salt 0.3% salt 0.1%
potassium sorbate 1.7% garlic flavor preparation 0.05% lactic acid
as pH-regulating agent flavors
[0141] A serving of 20 g of the cream cheese style spread contained
1 g plant sterols (as plant stanols) and 1.5 g protein hydrolysate
(as soy protein hydrolysate).
Example 15
[0142] Milk Alternative with Protein Hydrolysate/Plant
Sterol/Emulsifier Complex.
[0143] Protein hydrolysate/plant sterol/emulsifier complex from
example 8 was used in production of vanilla flavored milk
alternative.
[0144] Ingredients
TABLE-US-00006 5 l soymilk (2% fat) 94.5 g complex from example 8
vanilla flavoring
[0145] The soy protein hydrolysate/plant sterol/emulsifier complex
from example 8 was added under vigorous stirring to soymilk having
a temperature of 70.degree. C. and homogenized. 2 dl serving of the
soymilk contained 0.9 g of plant sterols and 1.6 g of soy protein
hydrolysate.
Example 16
[0146] Fruit Drink with Protein Hydrolysate/Plant Sterol/Emulsifier
Complex.
[0147] The protein hydrolysate/plant sterol/emulsifier complex from
example 8 was used in preparation of fruit drink of the following
ingredients:
TABLE-US-00007 200 g fruit juice concentrate (orange, pineapple,
passion fruit, guava, mango) 776 g water 8.5 g fructose 1.5 g
calcium lactate 14 g complex from example 8
[0148] The fruit juice concentrate, water, fructose and calcium
lactate were mixed together and heated to 70.degree. C. The soy
protein hydrolysate/plant sterol/emulsifier complex from example 8
was added under vigorous stirring to the drink and homogenized.
[0149] Two glasses (a 2 dl) of the drink contained 1.3 g of plant
sterols and 2.4 g of protein hydrolysate.
Example 17
[0150] Lipid Profile Improvement Obtained by Using a Composition of
the Present Invention.
[0151] The lipid profile improving effect of a composition
containing plant sterols (as plant stanol fatty acid ester) and
protein hydrolysate/emulsifier complex (as soy protein
hydrolysate/lysolecithin complex from example 2) was studied by
using LDL-receptor deficient female mice as test animals.
[0152] The aim of the test was to study the serum total cholesterol
and triglyceride lowering effect that could be obtained by using
only a small amount of plant sterols (0.5%, as sterol equivalents)
as part of a atherogenic diet containing protein hydrolysate. The
protein hydrolysate was administered in an amount to yield the
maximum cholesterol lowering effect that could be obtained by using
the hydrolysate in this animal model and still not compromising the
nutritional needs and growth of the mice. The suitable amount of
the hydrolysate was found to be 1/2 (as N equivalents) of the total
protein content of the diet.
[0153] The animals were assigned into 4 groups (n=8-10 in each
group) and fed experimental diets for 8 weeks.
[0154] All the experimental diets were formulated to contain 20%
protein. In the control group and in the test group 2, the sole
protein source was casein (88% purity). In the test groups 3 and 4,
the protein source was half casein and half (as N equivalents) the
protein hydrolysate/emulsifier complex. Test groups 2 and 4
contained 0.84% plant stanol fatty acid ester (0.5% as sterol
equivalents). Control group and test group 3 contained 0.35%
rapeseed oil to bring the equal amount of calories and equal fatty
acid composition, compared to the test groups getting plant stanol
ester.
[0155] Experimental diets and results:
TABLE-US-00008 Group 3 (protein Group 4 Group 2 hydrolysate/ (plant
stanol ester + Group 1 (plant stanol emulsifier protein
hydrolysate/ (control) ester) complex) emulsifier complex) % of
total % of total % of total % of total Ingredients Diet premix*
58.72 58.22 55.44 54.95 Cocoa butter 17.96 17.96 17.97 17.97
Rapeseed oil 0.35 0.00 0.35 0.00 Plant stanol ester 0.00 0.84 0.00
0.84 Casein powder 22.72 22.73 11.36 11.37 Soy protein hydrolysate/
0.00 0.00 14.62 14.62 lysolecithin complex, from example 2
Cholesterol 0.250 0.250 0.250 0.250 Results Serum total cholesterol
20.6 14.7 15.5 11.4 (mmol/l), mean Change (%) compared to -- -28.5
-25.0 -44.5 control group Serum triglycerides 2.2 2.4 1.9 1.3
(mmol/l), mean Change (%) compared to +8.8 -10.9 -39.1 control
group *Clinton/Cybulsky (D12106px, without protein), from Research
Diets Inc.
[0156] As shown in the table, both plant stanol ester and protein
hydrolysate/emulsifier complex had serum cholesterol lowering
effect (groups 2 and 3, respectively). A composition of the present
invention (group 4), the combination of plant sterols and protein
hydrolysate/emulsifier complex, had even enhanced cholesterol
lowering effect compared to the plant sterols (group 2) or protein
hydrolysate/emulsifier complex (group 3) alone and compared to what
was expected for the combination. Thus even when the maximal amount
of protein hydrolysate was used in the feed addition of plant
sterols effectively further reduced serum total cholesterol
levels
[0157] Protein hydrolysate/emulsifier complex (group 3) had serum
triglyceride reducing effect, whereas the serum triglyceride level
was somewhat raised in the group 2 receiving plant sterols.
Surprisingly, the combination of plant sterols and protein
hydrolysate/emulsifier complex had a strong synergistic
triglyceride lowering effect.
[0158] By combining plant sterols and protein hydrolysate,
according to the present invention, remarkable synergistic lipid
profile improvements could be seen.
* * * * *