Note: Descriptions are shown in the official language in which they were submitted.
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DESCRIPTION
NUTRITIONAL COMPOSITIONS COMPRISING LACTOBACILLUS RHAMNOSUS GG,
AS WELL AS DIETARY BUTYRATE AND/OR A COMPOUND FOR STIMULATING
FORMATION OF ENDOGENOUS BUTYRATE
TECHNICAL FIELD
[0001] The present disclosure relates generally to nutritional compositions
comprising a
combination of polydextrose ("PDX"), galacto-oligosaccharides ("GOS"), and
Lactobacillus rhamnosus GG ("LGG") or compositions comprising LGG and a source
of
dietary butyrate. The nutritional compositions are suitable for administration
to pediatric
subjects. Further, disclosed are methods for reducing risk for allergic
sensitization/prevention of allergic diseases, accelerating tolerance to cow's
milk
allergy and the dietary management of food allergy via administering the
compositions disclosed herein. The disclosed nutritional compositions may
provide
additive and or/synergistic beneficial health effects.
BACKGROUND ART
[0002] Food allergies, such as allergy to cow's milk protein, soy protein,
rice protein and
peanuts, are being recognized as an increasing problem. Cow's milk protein
allergy
("CMA") is the most common food allergy in early childhood and affects 2-3% of
young
children with a range of immunoglobulin (Ig-E) and non lg-E mediated
syndromes.
Food allergies continue to be a growing health concern with an increasing
prevalence
and severity, potential increase of atopic disease in later life, risk of
persistence, and
functional gastrointestinal disorders. Thus, there is a strong need to develop
effective
methods for supporting resistance to such allergies.
[0003] Typically, the first step of treatment of CMA is the rapid resolution
of symptoms,
with elimination of cow's milk protein from the diet being the only proven
treatment.
While certain hydrolyzed protein formulas have been used to avoid CMA, there
is a
need for nutritional compositions, especially for infants, including
components that are
capable of further accelerating tolerance acquisition to cow's milk.
[0004] Accordingly, provided herein are nutritional compositions that
accelerate
tolerance to cow's milk allergy. Further provided are compositions for the
dietary
management of food allergies.
DISCLOSURE OF THE INVENTION
[0005] Briefly, the present disclosure is directed, in an embodiment, to a
nutritional
composition that includes a synbiotic combination of a prebiotic composition
comprising PDX and GOS in combination with LGG. Additionally, the disclosure
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provides, in an embodiment, a nutritional composition comprising dietary
butyrate and
LGG. In some embodiments, the nutritional compositions include a component
which
can stimulate the production of endogenous short chain fatty acid ("SCFA")
including
butyrate in the human gut. In some embodiments, the dietary butyrate may be
provided directly by nutrition in the form of encapsulated butyrate or
enriched lipid
fractions from milk.
[0006] The present disclosure further provides methods for accelerating
tolerance to
cow's milk allergy in a target subject via administering the nutritional
compositions
disclosed herein to the target subject. Further provided are methods for the
dietary
management of allergy, such as cow's milk allergy, via administering the
nutritional
composition disclosed herein to the target subject.
[0007] It is to be understood that both the foregoing general description and
the
following detailed description present embodiments of the disclosure and are
intended
to provide an overview or framework for understanding the nature and character
of
the disclosure as it is claimed. The description serves to explain the
principles and
operations of the claimed subject matter. Other and further features and
advantages
of the present disclosure will be readily apparent to those skilled in the art
upon a
reading of the following disclosure.
BEST MODE FOR CARRYING OUT THE INVENTION
[0008] Reference now will be made in detail to the embodiments of the present
disclosure, one or more examples of which are set forth herein below. Each
example is
provided by way of explanation of the nutritional composition of the present
disclosure
and is not a limitation. In fact, it will be apparent to those skilled in the
art that various
modifications and variations can be made to the teachings of the present
disclosure
without departing from the scope of the disclosure. For instance, features
illustrated or
described as part of one embodiment, can be used with another embodiment to
yield
a still further embodiment.
[0009] Thus, it is intended that the present disclosure covers such
modifications and
variations as come within the scope of the appended claims and their
equivalents.
Other objects, features and aspects of the present disclosure are disclosed in
or are
apparent from the following detailed description. It is to be understood by
one of
ordinary skill in the art that the present discussion is a description of
exemplary
embodiments only and is not intended as limiting the broader aspects of the
present
disclosure.
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[0010] The present disclosure relates generally to nutritional compositions
comprising
dietary butyrate and a probiotic comprising LGG. In some embodiments, provided
are
nutritional compositions comprising a prebiotic comprising PDX and GOS and
LGG.
Additionally, the disclosure relates to methods for accelerating tolerance to
cow's milk
allergy or methods for the dietary management of allergy in target subjects.
[0011] "Allergy" as used herein is defined as an "abnormal hypersensitivity to
a
substance which is normally tolerated and generally considered harmless."
There are
two basic phases involved with the allergic response. The first stage involves
the
development of the early phase of an immediate-type hypersensitivity response
to
allergens. The first time an allergen meets the immune system, no allergic
reaction
occurs. Instead, the immune system prepares itself for future encounters with
the
allergen. Macrophages, which are scavenger cells, surround and break up the
invading allergen. The macrophages then display the allergen fragments on
their cell
walls to T lymphocytes, which are the main orchestrators of the body's immune
reaction. This cognitive signal plus several non-cognitive signals (e.g.
cytokines)
activate the nciive T-cells and instruct the T-cell differentiation into T-
cell effector
subpopulations. The key players in the allergic cascade are T-cells of the Th-
2
phenotype (TH-2). TH-2 type T-cells are characterized by the secretion of
several
cytokines including interleukin-4 (IL-4), IL-5 and IL-13. The cytokines IL-4
and IL-13 then
activate B lymphocytes which produce antibodies of the subclass E (IgE). IgE
antibodies are directed against the particular allergen. The interaction of
specific IgE
antibodies on the surface of effector cells (mast cells and basophils) with an
allergen
triggers the early phase of immediate type hypersensitivity responses.
[0012] This mast cell activation usually occurs within minutes after the
second exposure
to an allergen. IgE antibodies on mast cells, constructed during the
sensitization phase,
recognize the allergen and bind to the invader. Once the allergen is bound to
the
receptor, granules in the mast cells release their contents. These contents,
or
mediators, are proinflammatory substances such as histamine, platelet-
activating
factor, prostaglandins, cytokines and leukotrienes. These mediators actually
trigger the
allergy attack. Histamine stimulates mucus production and causes redness,
swelling,
and inflammation. Prostaglandins constrict airways and enlarge blood vessels.
[0013] The second phase of the allergic immune response is characterized by
infiltration
of inflammatory cells, such as eosinophils, into the airways after an allergen
exposure.
An important linkage between sensitization and inflammation is represented by
T-cells
that secrete mediators not only involved in IgE synthesis, but also
responsible for
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eosinophil recruitment, activation and survival. The tissue mast cells and
neighboring
cells produce chemical messengers that signal circulating basophils,
eosinophils, and
other cells to migrate into that tissue and help fight the foreign material.
Eosinophils
secrete chemicals of their own that sustain inflammation, cause tissue damage,
and
recruit yet more immune cells. This phase can occur anywhere between several
hours
and several days after the allergen exposure and can last for hours and even
days.
[0014] "Nutritional composition" means a substance or formulation that
satisfies at least
a portion of a subject's nutrient requirements. The terms "nutritional(s)",
"nutritional
formula(s)", "enteral nutritional(s)", and "nutritional supplement(s)" are
used as non-
limiting examples of nutritional composition(s) throughout the present
disclosure.
Moreover, "nutritional composition(s)" may refer to liquids, powders, gels,
pastes, solids,
concentrates, suspensions, or ready-to-use forms of enteral formulas, oral
formulas,
formulas for infants, formulas for pediatric subjects, formulas for children,
growing-up
milks and/or formulas for adults.
[0015] "Pediatric subject" means a human less than 13 years of age. In some
embodiments, a pediatric subject refers to a human subject that is between
birth and
8 years old. In other embodiments, a pediatric subject refers to a human
subject
between 1 and 6 years of age. In still further embodiments, a pediatric
subject refers to
a human subject between 6 and 12 years of age. The term "pediatric subject"
may
refer to infants (preterm or fullterm) and/or children, as described below.
[0016] "Infant" means a human subject ranging in age from birth to not more
than one
year and includes infants from 0 to 12 months corrected age. The phrase
"corrected
age" means an infant's chronological age minus the amount of time that the
infant
was born premature. Therefore, the corrected age is the age of the infant if
it had
been carried to full term. The term infant includes low birth weight infants,
very low
birth weight infants, and preterm infants. "Preterm" means an infant born
before the
end of the 37th week of gestation. "Full term" means an infant born after the
end of the
37th week of gestation.
[0017] "Child" means a subject ranging in age from 12 months to about 13
years. In
some embodiments, a child is a subject between the ages of 1 and 12 years old.
In
other embodiments, the terms "children" or "child" refer to subjects that are
between
one and about six years old, or between about seven and about 12 years old. In
other
embodiments, the terms "children" or "child" refer to any range of ages
between 12
months and about 13 years.
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[0018] "Infant formula" means a composition that satisfies at least a portion
of the
nutrient requirements of an infant. In the United States, the content of an
infant formula
is dictated by the federal regulations set forth at 21 C.F.R. Sections 100,
106, and 107.
[0019] The term "medical food" refers enteral compositions that are formulated
or
intended for the dietary management of a disease or disorder. A medical food
may
be a food for oral ingestion or tube feeding (nasogastric tube), may be
labeled for the
dietary management of a specific medical disorder, disease or condition for
which
there are distinctive nutritional requirements, and may be intended to be used
under
medical supervision.
[0020] The term "peptide" as used herein describes linear molecular chains of
amino
acids, including single chain molecules or their fragments. The peptides
described
herein include no more than 50 total amino acids. Peptides may further form
oligomers
or multimers consisting of at least two identical or different molecules.
Furthermore,
peptidomimetics of such peptides where amino acid(s) and/or peptide bond(s)
have
been replaced by functional analogs are also encompassed by the term
"peptide".
Such functional analogues may include, but are not limited to, all known amino
acids
other than the 20 gene-encoded amino acids such as selenocysteine.
[0021] The term "peptide" may also refer to naturally modified peptides where
the
modification is effected, for example, by glycosylation, acetylation,
phosphorylation
and similar modification which are well known in the art. In some embodiments,
the
peptide component is distinguished from a protein source also disclosed
herein.
Further, peptides may, for example, be produced recombinantly, semi-
synthetically,
synthetically, or obtained from natural sources such as after hydrolysation of
proteins,
including but not limited to casein, all according to methods known in the
art.
[0022] The term "molar mass distribution" when used in reference to a
hydrolyzed
protein or protein hydrolysate pertains to the molar mass of each peptide
present in
the protein hydrolysate. For example, a protein hydrolysate having a molar
mass
distribution of greater than 500 Daltons means that each peptide included in
the
protein hydrolysate has a molar mass of at least 500 Da!tons. Accordingly, in
some
embodiments, the peptides disclosed in Table 1 and Table 2 are derived from a
protein
hydrolysate having a molar mass distribution of greater than 500 Da!tons. To
produce a
protein hydrolysate having a molar mass distribution of greater than 500
Da!tons, a
protein hydrolysate may be subjected to certain filtering procedures or any
other
procedure known in the art for removing peptides, amino acids, and/or other
proteinaceous material having a molar mass of less than 500 Da!tons. For the
purposes
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of this disclosure, any method known in the art may be used to produce the
protein
hydrolysate having a molar mass distribution of greater than 500 Dalton.
[0023] The term "protein equivalent" or "protein equivalent source" includes
any
protein source, such as soy, egg, whey, or casein, as well as non-protein
sources, such
as peptides or amino acids. Further, the protein equivalent source can be any
used in
the art, e.g., nonfat milk, whey protein, casein, soy protein, hydrolyzed
protein,
peptides, amino acids, and the like. Bovine milk protein sources useful in
practicing the
present disclosure include, but are not limited to, milk protein powders, milk
protein
concentrates, milk protein isolates, nonfat milk solids, nonfat milk, nonfat
dry milk, whey
protein, whey protein isolates, whey protein concentrates, sweet whey, acid
whey,
casein, acid casein, caseinate (e.g. sodium caseinate, sodium calcium
caseinate,
calcium caseinate), soy bean proteins, and any combinations thereof. The
protein
equivalent source can, in some embodiments comprise hydrolyzed protein,
including
partially hydrolyzed protein and extensively hydrolyzed protein. The protein
equivalent
source may, in some embodiments, include intact protein. More particularly,
the
protein source may include a) about 20% to about 80% of the peptide component
described herein, and b) about 20% to about 80 % of an intact protein, a
hydrolyzed
protein, or a combination thereof.
[0024] The term "protein equivalent source" also encompasses free amino acids.
In
some embodiments, the amino acids may comprise, but are not limited to,
histidine,
isoleucine, leucine, lysine, methionine, cysteine, phenylalanine, tyrosine,
threonine,
tryptophan, valine, alanine, arginine, asparagine, aspartic acid, glutamic
acid,
glutamine, glycine, proline, serine, carnitine, taurine and mixtures thereof.
In some
embodiments, the amino acids may be branched chain amino acids. In certain
other
embodiments, small amino acid peptides may be included as the protein
component
of the nutritional composition. Such small amino acid peptides may be
naturally
occurring or synthesized.
[0025] "Fractionation procedure" includes any process in which a certain
quantity of a
mixture is divided up into a number of smaller quantities known as fractions.
The
fractions may be different in composition from both the mixture and other
fractions.
Examples of fractionation procedures include but are not limited to, melt
fractionation,
solvent fractionation, supercritical fluid fractionation and/or combinations
thereof.
[0026] "Milk fat globule membrane" includes components found in the milk fat
globule
membrane including but not limited to milk fat globule membrane proteins such
as
Mucin 1, Butyrophilin, Adipophilin, CD36, CD14, Lactadherin (PAS6/7), Xanthine
oxidase
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and Fatty Acid binding proteins etc. Additionally, "milk fat globule membrane"
may
include phospholipids, cerebrosides, gangliosides, sphingomyelins, and/or
cholesterol.
[0027] The term "growing-up milk" refers to a broad category of nutritional
compositions intended to be used as a part of a diverse diet in order to
support the
normal growth and development of a child between the ages of about 1 and about
6
years of age.
[0028] "Milk" means a component that has been drawn or extracted from the
mammary gland of a mammal. In some embodiments, the nutritional composition
comprises components of milk that are derived from domesticated ungulates,
ruminants or other mammals or any combination thereof.
[0029] "Nutritionally complete" means a composition that may be used as the
sole
source of nutrition, which would supply essentially all of the required daily
amounts of
vitamins, minerals, and/or trace elements in combination with proteins,
carbohydrates,
and lipids. Indeed, "nutritionally complete" describes a nutritional
composition that
provides adequate amounts of carbohydrates, lipids, essential fatty acids,
proteins,
essential amino acids, conditionally essential amino acids, vitamins, minerals
and
energy required to support normal growth and development of a subject.
[0030] A nutritional composition that is "nutritionally complete" for a full
term infant will,
by definition, provide qualitatively and quantitatively adequate amounts of
all
carbohydrates, lipids, essential fatty acids, proteins, essential amino acids,
conditionally
essential amino acids, vitamins, minerals, and energy required for growth of
the full
term infant.
[0031] A nutritional composition that is "nutritionally complete" for a child
will, by
definition, provide qualitatively and quantitatively adequate amounts of all
carbohydrates, lipids, essential fatty acids, proteins, essential amino acids,
conditionally
essential amino acids, vitamins, minerals, and energy required for growth of a
child.
[0032] "Exogenous butyrate" or "dietary butyrate" each refer to butyrate or
butyrate
derivatives which are intentionally included in the nutritional composition of
the present
disclosure itself, rather than generated in the gut.
[0033] "Endogenous butyrate" or "butyrate from endogenous sources" each refer
to
butyrate present in the gut as a result of ingestion of the disclosed
composition that is
not added as such, but is present as a result of other components or
ingredients of the
composition; the presence of such other components or ingredients of the
composition
stimulates butyrate production in the gut.
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[0034] The term "cow's milk allergy" describes a food allergy, i.e. an immune
adverse
reaction to one or more of the proteins contained in cow's milk in a human
subject. The
principal symptoms are gastrointestinal, dermatological, and respiratory
symptoms.
These can translate into skin rashes, hives, vomiting, diarrhea, constipation
and distress.
The clinical spectrum extends to diverse disorders: anaphylactic reactions,
atopic
dermatitis, wheeze, infantile colic, gastro esophageal reflux disease (GERD),
esophagitis, colitis gastroenteritis, headache/migraine and constipation.
[0035] "Probiotic" means a microorganism with low or no pathogenicity that
exerts a
beneficial effect on the health of the host.
[0036] The term "non-viable probiotic" means a probiotic wherein the metabolic
activity or reproductive ability of the referenced probiotic has been reduced
or
destroyed. More specifically, "non-viable" or "non-viable probiotic" means non-
living
probiotic microorganisms, their cellular components and/or metabolites
thereof. Such
non-viable probiotics may have been heat-killed or otherwise inactivated. The
"non-
viable probiotic" does, however, still retain, at the cellular level, its cell
structure or
other structure associated with the cell, for example exopolysaccharide and at
least a
portion its biological glycol-protein and DNA/RNA structure and thus retains
the ability
to favorably influence the health of the host. Contrariwise, the term "viable"
refers to
live microorganisms. As used herein, the term "non-viable" is synonymous with
"inactivated".
[0037] "Prebiotic" means a non-digestible food ingredient that beneficially
affects the
host by selectively stimulating the growth and/or activity of one or a limited
number of
bacteria in the digestive tract that can improve the health of the host.
[0038] The nutritional composition of the present disclosure may be
substantially free of
any optional or selected ingredients described herein, provided that the
remaining
nutritional composition still contains all of the required ingredients or
features described
herein. In this context, and unless otherwise specified, the term
"substantially free"
means that the selected composition may contain less than a functional amount
of the
optional ingredient, typically less than 0.1% by weight, and also, including
zero percent
by weight of such optional or selected ingredient.
[0039] All percentages, parts and ratios as used herein are by weight of the
total
composition, unless otherwise specified.
[0040] All references to singular characteristics or limitations of the
present disclosure
shall include the corresponding plural characteristic or limitation, and vice
versa, unless
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otherwise specified or clearly implied to the contrary by the context in which
the
reference is made.
[0041] All combinations of method or process steps as used herein can be
performed in
any order, unless otherwise specified or clearly implied to the contrary by
the context in
which the referenced combination is made.
[0042] The methods and compositions of the present disclosure, including
components
thereof, can comprise, consist of, or consist essentially of the essential
elements and
limitations of the embodiments described herein, as well as any additional or
optional
ingredients, components or limitations described herein or otherwise useful in
nutritional
compositions.
[0043] As used herein, the term "about" should be construed to refer to both
of the
numbers specified as the endpoint(s) of any range. Any reference to a range
should
be considered as providing support for any subset within that range.
[0044] The present disclosure is directed to nutritional compositions
including butyrate
and LGG. Non-limiting examples of butyrate for use herein include butyric
acid,
butyrate salts, glycerol esters of butyric acid, and combinations thereof. The
nutritional
compositions may further include a carbohydrate source, a protein source, and
a fat
or lipid source. In some embodiments, the nutritional compositions may include
a
component capable of stimulating endogenous butyrate production; in other
embodiments, the nutritional compositions may include both dietary and
endogenous
butyrate.
[0045] The benefit to providing both exogenous and endogenous butyrate is
accelerated tolerance acquisition towards cow's milk. Additionally, the
benefit to
providing both exogenous and endogenous butyrate together with Lactobacillus
rhamnoses GG ("LGG") is accelerated tolerance acquisition toward cow's milk.
Conventional dietary management of cow's milk allergy includes the use of
formulations containing protein hydrolysates and amino acids rather than
intact
proteins. However, the inclusion of certain probiotics, such as LGG in
combination with
butyrate, either endogenous or exogenous butyrate, can contribute to
accelerated
tolerance acquisition towards cow's milk.
[0046] In some embodiments, the nutritional composition includes a source of
dietary
butyrate that is present in an amount of from about 5 g/100 Kcal to about 500
g/100
kcal. In some embodiments, the nutritional composition includes a source of
dietary
butyrate that is present in an amount of from about 15 g/100 Kcal to about 450
g/100
kcal. In some embodiments, the nutritional composition includes a source of
dietary
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butyrate that is present in an amount of from about 20 g/100 Kcal to about 400
g/100
kcal. In some embodiments, the nutritional composition includes a source of
dietary
butyrate that is present in an amount of from about 25 g/100 Kcal to about 350
g/100
kcal. In some embodiments, the nutritional composition includes a source of
dietary
butyrate that is present in an amount of from about 30 g/100 Kcal to about 280
g/100
kcal.
[0047] In some embodiments, the nutritional composition includes from about
0.01 g to
about 10 g of dietary butyrate per 100 g of total fat in the nutritional
composition. In
some embodiments, the nutritional composition includes from about 0.1 g to
about 8 g
of dietary butyrate per 100 g of total fat in the nutritional composition. In
some
embodiments, the nutritional composition includes from about 0.4 g to about 7
g of
dietary butyrate per 100 g of total fat in the nutritional composition. In
some
embodiments, the nutritional composition includes from about 0.7 g to about
6.5 g of
dietary butyrate per 100 g of total fat in the nutritional composition. In
some
embodiments, the nutritional composition includes from about 1.2 g to about
5.1 g of
dietary butyrate per 100 g of total fat in the nutritional composition.
[0048] In some embodiments the dietary butyrate is provided by one or more of
the
following: butyric acid; butyrate salts, including sodium butyrate, potassium
butyrate,
calcium butyrate, and/or magnesium butyrate; glycerol esters of butyric acid;
and/or
corresponding mixtures and corresponding salts of pharmaceutically acceptable
bases or acids, pure diastereoisomeric forms and enantiomeric forms or
mixtures
thereof.
[0049] The dietary butyrate can be supplied by any suitable source known in
the art.
Non-limiting sources of dietary butyrate includes animal source fats and
derived
products, such as but not limited to milk, milk fat, butter, buttermilk,
butter serum,
cream; microbial fermentation derived products, such as but not limited to
yogurt and
fermented buttermilk; and plant source derived seed oil products, such as
pineapple
and/or pineapple oil, apricot and/or apricot oil, barley, oats, brown rice,
bran, green
beans, legumes, leafy greens, apples, kiwi, oranges. In some embodiments, the
dietary
butyrate is synthetically produced. In embodiments where the dietary butyrate
is
synthetically produced, the chemical structure of the dietary butyrate may be
modified as necessary. Further, the dietary butyrate produced synthetically
can be
purified by any means known in the art to produce a purified dietary butyrate
additive
that can be incorporated into the nutritional compositions disclosed herein.
The dietary
butyrate may be provided by dairy lipids and/or triglyceride bound forms of
butyrate.
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[0050] In some embodiments, the dietary butyrate may be provided in an
encapsulated form. In certain embodiments, the encapsulation of the dietary
butyrate
may provide for longer shelf-stability and may provide for improved
organoleptic
properties of the nutritional composition. For example, in some embodiments,
the
dietary butyrate may be encapsulated or coated by the use of, or combination
of, fat
derived materials, such as mono- and di-glycerides; sugar and acid esters of
glycerides; phospholipids; plant, animal and microbial derived proteins and
hydrocolloids, such as starches, maltodextrins, gelatin, pectins, glucans,
caseins, soy
proteins, and/or whey proteins.
[0051] The dietary butyric acid may also be provided in a coated form. For
example,
coating certain glycerol esters of butyric acids and/or amide derivatives of
butyric
acids with fat derived materials, such as mono- and di-glycerides; sugar and
acid
esters of glycerides; phospholipids; plant, animal and microbial derived
proteins and
hydrocolloids, such as starches, maltodextrins, gelatin, pectins, glucans,
caseins, soy
proteins, and/or whey proteins may improve the shelf-stability of the dietary
butyrate
and may further improve the overall organoleptic properties of the nutritional
composition.
[0052] In certain embodiments, the dietary butyrate comprises alkyl, and or
glycerol
esters of butyric acid. Glycerol esters of butyric acid may offer minimal
complexity
when formulated and processed in the nutritional composition. Additionally,
glycerol
esters of butyric acid may improve the shelf life of the nutritional
composition including
dietary butyrate an may further have a low impact on the sensory attributes of
the
finished product.
[0053] In some embodiments, the dietary butyrate may comprise butyrate salts,
for
example, sodium butyrate, potassium butyrate, calcium butyrate, magnesium
butyrate, and combinations thereof. In some embodiments, the use of selected
dietary butyrate salts may improve intestinal health when provided to target
subjects.
In certain embodiments, dietary butyrate comprises a suitable butyrate salt
that has
been coated with one or more fats or lipids. In certain embodiments wherein
the
dietary butyrate comprises a fat coated butyrate salt, the nutritional
composition may
be a dry-powdered composition into which the dietary butyrate is incorporated.
[0054] In some embodiments, the dietary butyrate may comprise any of the
butyrate
compounds disclosed herein that are formulated to be in complex form with
chitosan
or one or cyclodextrins. For example, cyclodextrins are cyclic
oligosaccharides
composed of six (a-cyclodextrin), seven (13-cyclodextrin), or eight (gamma-
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cyclodextrin) units of a-1,4-glucopyranose. Cyclodextrins are further
characterized by
a hydrophilic exterior surface and a hydrophobic core. Without being bound by
any
particular theory, the aliphatic butyrate chain would form a complex with the
cyclodextrin core, thus increasing its molecular weight and, thus, reducing
the volatility
of the butyrate compound. Accordingly, the bioavailability of dietary butyrate
may be
improved when the dietary butyrate includes butyrate compounds in complex form
with one or more cyclodextrins. Further, cyclodextrins are bulky hydrophobic
molecules
that are resistant to stomach acid as well as gastrointestinal enzymes, thus
administration of the butyrate-cyclodextrin complex as described herein would
promote absorption of the dietary butyrate in the small intestines.
[0055] In some embodiments the dietary butyrate is provided from an enriched
lipid
fraction derived from milk. For example, bovine milk fat has a butyric acid
content that
may be 20 times higher than the butyric acid content in human milk fat.
Furthermore,
among the short chain fatty acids ("SCFAs") present in human milk, i.e. fatty
acids
having a carbon chain length from 4 to 12, butyric acid (C4) is one of the
most
predominant in bovine milk. As such, bovine milk fat and/or enriched fractions
of
bovine milk fat may be included in a nutritional composition to provide
dietary
butyrate.
[0056] In embodiments where the dietary butyrate is provided by an enriched
lipid
fraction derived from milk the enriched lipid fraction derived from milk may
be
produced by any number of fractionation techniques. These techniques include
but
are not limited to melting point fractionation, organic solvent fractionation,
super
critical fluid fractionation, and any variants and combinations thereof.
[0057] Furthermore, mixtures that may be subjected to the fractionation
procedures to
produce the enriched lipid fraction include, but are not limited to, bovine
whole milk,
bovine cream, caprine milk, ovine milk, yak milk, and/or mixtures thereof. In
a
preferred embodiment the milk mixture used to create the enriched lipid
fraction is
bovine milk.
[0058] In addition to providing dietary butyrate, the enriched lipid fraction
may
comprise an one of the following ingredients: saturated fatty acids; trans-
fatty acids;
branched-chain fatty acids ("BCFAs"), including odd-branched chain fatty acids
("OBCFAs"); conjugated linoleic acid ("CLA"); monounsaturated fatty acids;
polyunsaturated fatty acids; cholesterol; phospholipids; and milk fat globule
membrane, including milk fat globule membrane protein.
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[0059] In some embodiments the enriched lipid fraction includes, per 100 Kcal,
one or
more of the following:
from about 0.1 g to 8.0 g of saturated fatty acids;
from about 0.2 g to 7.0 g trans-fatty acids;
from about 0.003 g to about 6.1 g branched-chain fatty acids;
from about 0.026 g to about 2.5 g conjugated linoleic acid;
from about 0.8 g to about 2.5 g monounsaturated fatty acids;
from about 2.3 g to about 4.4 g polyunsaturated fatty acids;
from about 100 mg to about 400 mg of cholesterol;
from about 50 mg to about 400 mg of phospholipids; and/or
from about 10 mg to about 500 mg of milk fat globule membrane.
[0060] The following example illustrates a milk fat fraction having an
enriched
concentration of butyric acid (C4) that may be produced by a fractionation
procedure.
Example 1
[0061] Illustrated below is a lipid profile of fractionated milk fat produced
by super
critical carbon extraction fractionation procedure and by melt-fractionation.
Milk Fat composition (g fatty acid /100 g TOTAL fatty acids)
Mel tFrac
AMF SCCO2
10C
C 4:0 3.9 6.0 4.7
C 6:0 2.5 3.3 2.9
C 8:0 1.4 1.9 1.8
C 10:0 3.1 3.9 3.8
C 12:0 4.2 4.1 4.8
C 14:0 11.4 12.2 10.9
C 14:1 1.1 1.0 1.3
C 15:0 1.1 1.0 0.9
C 16:0 29.4 29.6 22.3
C 16:1 1.9 1.4 2.2
C 17:0 0.6 0.5 0.4
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C 18:0 11.4 8.2 6.1
C 18:1, cis, co9 21.9 16.5 25.3
C 18:1, trans, co9 0.3 1.6 1.9
C 18:2, o.) 6 1.9 2.2 1.9
C 18:3, c,.) 3, a 0.6 0.4 0.6
C 20:0 0.0 0.1 0.1
C 20:1, co 9 0.1 0.1 0.2
Saturated 68.7 70.7 58.6
Unsaturated 27.8 23.1 33.3
AIV1F = anhydrous milk fat; SCCO2 = super-critical carbon dioxide fraction
(super olein).
MeltFrac = melt crystallization fraction separated at 10 C.
[0062] The nutritional composition of the present disclosure also includes at
least one
probiotic; in a preferred embodiment, the probiotic comprises LGG. In certain
other
embodiments, the probiotic may be selected from any other Lactobacillus
species,
Bifidobacterium species, Bifidobacterium Ion gum BB536 (BL999, ATCC: BAA-999),
Bifidobacterium longum AH1206 (NCIMB: 41382), Bifidobacterium breve AH1205
(NCIMB: 41387), Bifidobacterium infantis 35624 (NCIMB: 41003), and
Bifidobacterium
animalis subsp. lactis BB-12 (DSM No. 10140) or any combination thereof.
[0063] The amount of the probiotic may vary from about 1 x 104 to about 1.5 x
1012 cfu
of probiotic(s) per 100 kcal. In some embodiments the amount of probiotic may
be
from about 1 x 106 to about 1 x 109 cfu of probiotic(s) per 100 kcal. In
certain other
embodiments the amount of probiotic may vary from about 1 x 107 cfu/100 kcal
to
about 1 x 108 cfu of probiotic(s) per 100 kcal.
[0064] As noted, in a preferred embodiment, the probiotic comprises LGG. LGG
is a
probiotic strain isolated from healthy human intestinal flora. It was
disclosed in U.S.
Patent No. 5,032,399 to Gorbach, et at., which is herein incorporated in its
entirety, by
reference thereto. LGG is resistant to most antibiotics, stable in the
presence of acid
and bile, and attaches avidly to mucosal cells of the human intestinal tract.
It survives
for 1-3 days in most individuals and up to 7 days in 30% of subjects. In
addition to its
colonization ability, LGG also beneficially affects mucosal immune responses.
LGG is
deposited with the depository authority American Type Culture Collection
("ATCC")
under accession number ATCC 53103.
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[0065] In an embodiment, the probiotic(s) may be viable or non-viable. The
probiotics
useful in the present disclosure may be naturally-occurring, synthetic or
developed
through the genetic manipulation of organisms, whether such source is now
known or
later developed.
[0066] In some embodiments, the nutritional composition may include a source
comprising probiotic cell equivalents, which refers to the level of non-
viable, non-
replicating probiotics equivalent to an equal number of viable cells. The term
"non-
replicating" is to be understood as the amount of non-replicating
microorganisms
obtained from the same amount of replicating bacteria (cfu/g), including
inactivated
probiotics, fragments of DNA, cell wall or cytoplasmic compounds. In other
words, the
quantity of non-living, non-replicating organisms is expressed in terms of cfu
as if all the
microorganisms were alive, regardless whether they are dead, non-replicating,
inactivated, fragmented etc. In non-viable probiotics are included in the
nutritional
composition, the amount of the probiotic cell equivalents may vary from about
1 x 104
to about 1.5 x 1010 cell equivalents of probiotic(s) per 100 kcal. In some
embodiments
the amount of probiotic cell equivalents may be from about 1 x 106 to about 1
x 109 cell
equivalents of probiotic(s) per 100 kcal nutritional composition. In certain
other
embodiments the amount of probiotic cell equivalents may vary from about 1 x
107 to
about 1 x 108 cell equivalents of probiotic(s) per 100 kcal of nutritional
composition.
[0067] In some embodiments, the probiotic source incorporated into the
nutritional
composition may comprise both viable colony-forming units, and non-viable cell-
equivalents.
[0068] While, probiotics may be helpful in pediatric patients, the
administration of viable
bacteria to pediatric subjects, and particularly preterm infants, with
impaired intestinal
defenses and immature gut barrier function may not be feasible due to the risk
of
bacteremia. Therefore, there is a need for compositions that can provide the
benefits
of probiotics without introducing viable bacteria into the intestinal tract of
pediatric
subjects
[0069] While not wishing to be bound by theory, it is believed that a culture
supernatant
from batch cultivation of a probiotic, and in particular embodiments, LGG,
provides
beneficial gastrointestinal benefits. It is further believed that the
beneficial effects on
gut barrier function can be attributed to the mixture of components (including
proteinaceous materials, and possibly including (exo)polysaccharide materials)
that
are released into the culture medium at a late stage of the exponential (or
"log")
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phase of batch cultivation of LGG. The composition will be hereinafter
referred to as
"culture supernatant."
[0070] Accordingly, in some embodiments, the nutritional composition includes
a
culture supernatant from a late-exponential growth phase of a probiotic batch-
cultivation process. Without wishing to be bound by theory, it is believed
that the
activity of the culture supernatant can be attributed to the mixture of
components
(including proteinaceous materials, and possibly including (exo)polysaccharide
materials) as found released into the culture medium at a late stage of the
exponential
(or "log") phase of batch cultivation of the probiotic. The term "culture
supernatant" as
used herein, includes the mixture of components found in the culture medium.
The
stages recognized in batch cultivation of bacteria are known to the skilled
person.
These are the "lag," the "log" ("logarithmic" or "exponential"), the
"stationary" and the
"death" (or "logarithmic decline") phases. In all phases during which live
bacteria are
present, the bacteria metabolize nutrients from the media, and secrete (exert,
release)
materials into the culture medium. The composition of the secreted material at
a given
point in time of the growth stages is not generally predictable.
[0071] In an embodiment, a culture supernatant is obtainable by a process
comprising
the steps of (a) subjecting a probiotic such as LGG to cultivation in a
suitable culture
medium using a batch process; (b) harvesting the culture supernatant at a late
exponential growth phase of the cultivation step, which phase is defined with
reference to the second half of the time between the lag phase and the
stationary
phase of the batch-cultivation process; (c) optionally removing low molecular
weight
constituents from the supernatant so as to retain molecular weight
constituents above
5-6 kiloDaltons (kDa); (d) removing liquid contents from the culture
supernatant so as to
obtain the composition.
[0072] The culture supernatant may comprise secreted materials that are
harvested
from a late exponential phase. The late exponential phase occurs in time after
the mid
exponential phase (which is halftime of the duration of the exponential phase,
hence
the reference to the late exponential phase as being the second half of the
time
between the lag phase and the stationary phase). In particular, the term "late
exponential phase" is used herein with reference to the latter quarter portion
of the
time between the lag phase and the stationary phase of the LGG batch-
cultivation
process. In some embodiments, the culture supernatant is harvested at a point
in time
of 75% to 85% of the duration of the exponential phase, and may be harvested
at
about 5/6 of the time elapsed in the exponential phase.
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[0073] The culture supernatant is believed to contain a mixture of amino
acids, oligo-
and polypeptides, and proteins, of various molecular weights. The composition
is
further believed to contain polysaccharide structures and/or nucleotides.
[0074] In some embodiments, the culture supernatant of the present disclosure
excludes low molecular weight components, generally below 6 kDa, or even below
5
kDa. In these and other embodiments, the culture supernatant does not include
lactic
acid and/or lactate salts. These lower molecular weight components can be
removed, for example, by filtration or column chromatography.
[0075] The culture supernatant of the present disclosure can be formulated in
various
ways for administration to pediatric subjects. For example, the culture
supernatant can
be used as such, e.g. incorporated into capsules for oral administration, or
in a liquid
nutritional composition such as a drink, or it can be processed before further
use. Such
processing generally involves separating the compounds from the generally
liquid
continuous phase of the supernatant. This preferably is done by a drying
method, such
as spray-drying or freeze-drying (Iyophilization). Spray-drying is preferred.
In a preferred
embodiment of the spray-drying method, a carrier material will be added before
spray-drying, e.g., maltodextrin DE29.
[0076] The LGG culture supernatant of the present disclosure, whether added in
a
separate dosage form or via a nutritional product, will generally be
administered in an
amount effective in promoting gut regeneration, promoting gut maturation
and/or
protecting gut barrier function. The effective amount is preferably equivalent
to 1x104
to about lx1 012 cell equivalents of live probiotic bacteria per kg body
weight per day,
and more preferably 108-109 cell equivalents per kg body weight per day. In
other
embodiments, the amount of cell equivalents may vary from about 1 x 104 to
about 1.5
x 1010 cell equivalents of probiotic(s) per 100 Kcal. In some embodiments the
amount
of probiotic cell equivalents may be from about 1 x 106 to about 1 x 109 cell
equivalents
of probiotic(s) per 100 Kcal nutritional composition. In certain other
embodiments the
amount of probiotic cell equivalents may vary from about 1 x 107 to about 1 x
108 cell
equivalents of probiotic(s) per 100 Kcal of nutritional composition.
[0077] Without being bound by any theory, it is believed the disclosed
combination of
dietary butyrate and probiotic, especially LGG, provides a higher potential to
bring
allergic infants and children to a normal diet, fast management of CMA
manifestations,
and can accelerate the development of tolerance acquisition in infants
affected by
CMA. The unique combination of nutrients in the disclosed nutritional
composition(s) is
believed to be capable of providing novel and unexpected benefits for infants
and
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18
children. Moreover, the benefit of this nutritional composition is believed to
be
obtained during infancy, and also by including it as part of a diverse diet as
the child
continues to grow and develop.
[0078] In an embodiment, the nutritional composition further comprises a
component
for stimulating endogenous butyrate production. For example, in some
embodiments
the component for stimulating endogenous butyrate production comprises a
microbiota-stimulating component that is a prebiotic including both
polydextrose
("PDX") and galacto-oligosaccharides ("GOS"). A prebiotic component including
PDX
and GOS can enhance butyrate production by microbiota. Butyrate has epigenetic
(histone deacetylase inhibition activity) that results in regulatory responses
such as
generation of regulatory T-cells. In the context of cow s milk allergy, these
regulatory
responses may result in accelerated tolerance acquisition to cow's milk
protein.
[0079] In addition to PDX and GOS, the nutritional composition may also
contain one or
more other prebiotics which can exert additional health benefits, which may
include,
but are not limited to, selective stimulation of the growth and/or activity of
one or a
limited number of beneficial gut bacteria, stimulation of the growth and/or
activity of
ingested probiotic microorganisms, selective reduction in gut pathogens, and
favorable influence on gut short chain fatty acid profile. Such prebiotics may
be
naturally-occurring, synthetic, or developed through the genetic manipulation
of
organisms and/or plants, whether such new source is now known or developed
later.
Prebiotics useful in the present disclosure may include oligosaccharides,
polysaccharides, and other prebiotics that contain fructose, xylose, soya,
galactose,
glucose and mannose.
[0080] More specifically, prebiotics useful in the present disclosure include
PDX and
GOS, and can, in some embodiments, also include, PDX powder, lactulose,
lactosucrose, raffinose, gluco-oligosaccharide, inulin, fructo-oligosaccharide
(FOS),
isomalto-oligosaccharide, soybean oligosaccharides, lactosucrose, xylo-
oligosaccharide (XOS), chito-oligosaccharide, manno-oligosaccharide, aribino-
oligosaccharide, siallyl-oligosaccharide, fuco-oligosaccharide, and gentio-
oligosaccharides.
[0081] In an embodiment, the total amount of prebiotics present in the
nutritional
composition may be from about 1.0 g/L to about 10.0 g/L of the composition.
More
preferably, the total amount of prebiotics present in the nutritional
composition may be
from about 2.0 g/L and about 8.0 g/L of the composition. In some embodiments,
the
total amount of prebiotics present in the nutritional composition may be from
about
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0.01 g/100 Kcal to about 1.5 g/100 Kcal. In certain embodiments, the total
amount of
prebiotics present in the nutritional composition may be from about 0.15 g/100
Kcal to
about 1.5 g/100 Kcal. In some embodiments, the prebiotic component comprises
at
least 20% w/w PDX and GOS.
[0082] The amount of PDX in the nutritional composition may, in an embodiment,
be
within the range of from about 0.015 g/100 Kcal to about 1.5 g/100 Kcal. In
another
embodiment, the amount of polydextrose is within the range of from about 0.2
g/100
Kcal to about 0.6 g/100 Kcal. In some embodiments, PDX may be included in the
nutritional composition in an amount sufficient to provide between about 1.0
g/L and
10.0 g/L. In another embodiment, the nutritional composition contains an
amount of
PDX that is between about 2.0 g/L and 8.0 g/L. And in still other embodiments,
the
amount of PDX in the nutritional composition may be from about 0.05 g/100 Kcal
to
about 1.5 g/100 Kcal.
[0083] The prebiotic component also comprises GOS. The amount of GOS in the
nutritional composition may, in an embodiment, be from about 0.015 g/100 Kcal
to
about 1.0 g/100 Kcal. In another embodiment, the amount of GOS in the
nutritional
composition may be from about 0.2 g/100 Kcal to about 0.5 g/100 Kcal.
[0084] In a particular embodiment, GOS and PDX are supplemented into the
nutritional
composition in a total amount of at least about 0.015 g/100 Kcal or about
0.015 g/100
Kcal to about 1.5 g/100 Kcal. In some embodiments, the nutritional composition
may
comprise GOS and PDX in a total amount of from about 0.1 to about 1.0 g/100
Kcal.
[0085] In certain embodiments, the PDX- and GOS-containing prebiotic and
dietary
butyrate is incorporated into a nutritional composition that is an infant
formula.
Currently, many infant formulas are not formulated with dietary butyrate. One
reason
that infant formulas include little to no dietary butyrate is due to the
unpleasant
organoleptic properties exhibited by the nutritional composition when butyrate
compounds are incorporated into the nutritional composition. For example, many
butyrate compounds exhibit an odor that makes consuming the nutritional
composition
in which they are incorporated an unpleasant experience. Furthermore, the
pediatric
and infant population will not readily consume nutritional products having an
unpleasant odor, taste, and/or mouthfeel. Accordingly, there exists a need for
a
nutritional composition formulated for administration to a pediatric subject
or an infant
that provides butyrate in the gut yet does not have diminished organoleptic
properties.
The incorporation of a prebiotic to stimulate butyrate production by gut
microbiota
and certain dietary butyrate compounds disclosed herein, i.e. glycerol esters
of butyric
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acid, into pediatric and infant nutritional compositions will provide butyrate
while still
providing a pleasant sensory experience.
[0086] In some embodiments, the nutritional composition includes a protein
equivalent
source, wherein the protein equivalent source includes a peptide component
comprising SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO
24, SEQ
ID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO
60, and
SEQ ID NO 63. In some embodiments, the peptide component may comprise
additional peptides disclosed in Table 1. For example, the composition may
include at
least 10 additional peptides disclosed in Table 1. In some embodiments, 20% to
80% of
the protein equivalent source comprises the peptide component, and 20% to 80%
of
the protein equivalent source comprises an intact protein, a partially
hydrolyzed
protein, and combinations thereof. In some embodiments, the term additional
means
selecting different peptides than those enumerated.
[0087] In another embodiment, 1% to about 99% of the protein equivalent source
includes a peptide component comprising at least 3 peptides selected from the
group
consisting of SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO
24,
SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID
NO 60,
and SEQ ID NO 63, and at least 5 additional peptides selected from Table 1;
and
wherein 1% to 99% of the protein equivalent source comprises an intact
protein, a
partially hydrolyzed protein, or combinations thereof. In some embodiments,
20% to
80% of the protein equivalent source includes a peptide component comprising
at
least 3 peptides selected from the group consisting of SEQ ID NO 4, SEQ ID NO
13, SEQ
ID NO 17, SEQ ID NO 21, SEQ ID NO 24, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO
32, SEQ
ID NO 51, SEQ ID NO 57, SEQ ID NO 60, and SEQ ID NO 63, and at least 5
additional
peptides selected from Table 1; and wherein 20% to 80% of the protein
equivalent
source comprises an intact protein, a partially hydrolyzed protein, or
combinations
thereof.
[0088] Table 1 below identifies the amino acid sequences of the peptides that
may be
included in the peptide component of the present nutritional compositions.
TABLE 1
Seq.
ID Amino Acid Sequence
(aa)
1 Ala Ile Asn Pro Ser Lys Glu Asn
8
2 Ala Pro Phe Pro Glu
5
3 Asp Ile Gly Ser Glu Ser
6
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4 Asp Lys Thr Glu lie Pro Thr
7
Asp Met Glu Ser Thr 5
6 Asp Met Pro He
4
7 Asp Val Pro Ser
4
n/a Glu Asp Ile
3
n/a Glu Leu Phe
3
n/a Glu Met Pro
3
8 Glu Thr Ala Pro Val Pro Leo
7
9 Phe Pro Gly Pro lie Pro
6
Phe Pro Gly Pro Ile Pro Asn 7
11 Gly Pro Phe Pro
4
12 Gly Pro Ile Val
4
13 lie Gly Ser Glu Ser Thr Glu Asp Gin
9
14 lie Gly Ser Ser Ser Glu Glu Ser
8
Ile Gly Ser Ser Ser Glu Glu Ser Ala 9
16 Ile Asn Pro Ser Lys Glu
6
17 Ile Pro Asn Pro Ile
5
18 Ile Pro Asn Pro Ile Gly
6
19 Ile Pro Pro Leu Thr Gin Thr Pro Val
9
lie Thr Ala Pro 4
21 Ile Val Pro Asn
4
22 Lys His Gin Gly Leu Pro Gin
7
23 Leu Asp Val Thr Pro
5
24 Leu Glu Asp Ser Pro Glu
6
Leu Pro Leu Pro Leu 5
26 Met Glu Ser Thr Giu Val
6
27 Met His Gin Pro His Gin Pro Leu Pro Pro Thr
11
28 Asn Ala Val Pro lie
5
29 Asn Glu Val Glu Ala
5
n/a Asn Leu Leu
3
Asn Gin Glu Gin Pro Ile 6
31 Asn Val Pro Gly Giu
5
32 Pro Phe Pro Gly Pro Ile
6
33 Pro Gly Pro lie Pro Asn
6
34 Pro His Gin Pro Leu Pro Pro Thr
8
Pro lie Thr Pro Thr 5
36 Pro Asn Pro Ile
4
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37 Pro Asn Ser Leu Pro Gin
6
38 Pro Gin Leu Giu lie Val Pro Asn
8
39 Pro Gin Asn He Pro Pro Leu
7
40 Pro Val Leu Gly Pro Val
6
41 Pro Val Pro Gin
4
42 Pro Val Val Val Pro
5
43 Pro Val Val Val Pro Pro
6
44 Ser Ile Gly Ser Ser Ser Glu Glu Ser Ala Glu 11
45 Ser Ile Ser Ser Ser Glu Glu
7
46 Ser Ile Ser Ser Ser Glu Glu Ile Val Pro Asn 11
47 Ser Lys Asp Ile Gly Ser Glu
7
48 Ser Pro Pro Glu Ile Asn
6
49 Ser Pro Pro Giu lie Asn Thr
7
50 Thr Asp Ala Pro Ser Phe Ser
7
51 Thr Glu Asp Giu Leu
5
52 Vol Ala Thr Glu Glu Vol
6
53 Vol Leu Pro Vol Pro
5
54 Vol Pro Gly Glu
4
55 Vol Pro Gly Giu lie Vol
6
56 Vol Pro Ile Thr Pro Thr
6
57 Vol Pro Ser Giu
4
58 Vol Vol Pro Pro Phe Leu Gin Pro Glu
9
59 Vol Vol Vol Pro Pro
5
60 Tyr Pro Phe Pro Gly Pro
6
61 Tyr Pro Phe Pro Gly Pro Ile
Pro 8
62 Tyr Pro Phe Pro Gly Pro Ile Pro Asn
9
63 Tyr Pro Ser Gly Ala
5
64 Tyr Pro Vol Glu Pro
5
[0089] Table 2 below further identifies a subset of amino acid sequences from
Table 1
that may be included in the peptide component disclosed herein.
TABLE 2
Seq ID
Number Amino Acid Sequence (aa)
4 Asp Lys Thr Glu lie Pro Thr 7
13 lie Gly Ser Giu Ser Thr Giu Asp Gin 9
17 lie Pro Asn Pro lie Gly 6
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21 Ile Val Pro Asn 4
24 Leu Glu Asp Ser Pro Glu 6
30 Asn Gin Glu Gin Pro Ile 6
31 Asn Val Pro Giy Glu 5
32 Pro Phe Pro Gly Pro Ile 6
51 Thr Gil.) Asp Glu Leu 5
57 Val Pro Ser Glu 4
60 Tyr Pro Phe Pro Gly Pro 6
63 Tyr Pro Ser Giy Ala 5
[0090] In some embodiments, the peptide component may be present in the
nutritional
composition in an amount from about 0.2 g/100 Kcal to about 5.6 g/100 Kcal. In
other
embodiments the peptide component may be present in the nutritional
composition in
an amount from about 1 g/100 Kcal to about 4 g/100 Kcal. instill other
embodiments,
the peptide component may be present in the nutritional composition in an
amount
from about 2 g/100 Kcal to about 3 g/100 Kcal.
[0091] The peptide component disclosed herein may be formulated with other
ingredients in the nutritional composition to provide appropriate nutrient
levels for the
target subject. In some embodiments, the peptide component is included in a
nutritionally complete formula that is suitable to support normal growth.
[0092] The peptide component may be provided as an element of a protein
equivalent source. In some embodiments, the peptides identified in Tables 1
and 2,
may be provided by a protein equivalent source obtained from cow's milk
proteins,
including but not limited to bovine casein and bovine whey. In some
embodiments,
the protein equivalent source comprises hydrolyzed bovine casein or hydrolyzed
bovine whey. Accordingly, in some embodiments, the peptides identified in
Table 1
and Table 2 may be provided by a casein hydrolysate. Such peptides may be
obtained by hydrolysis or may be synthesized in vitro by methods know to the
skilled
person.
[0093] A non-limiting example of a method of hydrolysis is disclosed herein.
In some
embodiments, this method may be used to obtain the protein hydrolysate and
peptides of the present disclosure. The proteins are hydrolyzed using a
proteolytic
enzyme, Protease N. Protease N "Amano" is commercially available from Amano
Enzyme U.S.A. Co., Ltd., Elgin, Ill. Protease N is a proteolytic enzyme
preparation that is
derived from the bacterial species Bacillus subtilis. The protease powder is
specified as
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"not less than 150,000 units/g", meaning that one unit of Protease N is the
amount of
enzyme which produces an amino acid equivalent to 100 micrograms of tyrosine
for 60
minutes at a pH of 7Ø To produce the infant formula of the present
disclosure,
Protease N can be used at levels of about 0.5% to about 1.0% by weight of the
total
protein being hydrolyzed.
[0094] The protein hydrolysis by Protease N is typically conducted at a
temperature of
about 50 C. to about 60 C. The hydrolysis occurs for a period of time so as
to obtain a
degree of hydrolysis between about 4% and 10%. In a particular embodiment,
hydrolysis occurs for a period of time so as to obtain a degree of hydrolysis
between
about 6% and 9%. In another embodiment, hydrolysis occurs for a period of time
so as
to obtain a degree of hydrolysis of about 7.5%. This level of hydrolysis may
take
between about one half hour to about 3 hours.
[0095] A constant pH should be maintained during hydrolysis. In the method of
the
present disclosure, the pH is adjusted to and maintained between about 6.5 and
8. In a
particular embodiment, the pH is maintained at about 7Ø
[0096] In order to maintain the optimal pH of the solution of whey protein,
casein, water
and Protease N, a caustic solution of sodium hydroxide and/or potassium
hydroxide
can be used to adjust the pH during hydrolysis. If sodium hydroxide is used to
adjust the
pH, the amount of sodium hydroxide added to the solution should be controlled
to the
level that it comprises less than about 0.3% of the total solid in the
finished protein
hydrolysate. A 10% potassium hydroxide solution can also be used to adjust the
pH of
the solution to the desired value, either before the enzyme is added or during
the
hydrolysis process in order to maintain the optimal pH.
[0097] The amount of caustic solution added to the solution during the protein
hydrolysis can be controlled by a pH-stat or by adding the caustic solution
continuously
and proportionally. The hydrolysate can be manufactured by standard batch
processes or by continuous processes.
[0098] To better ensure the consistent quality of the protein partial
hydrolysate, the
hydrolysate is subjected to enzyme deactivation to end the hydrolysis process.
The
enzyme deactivation step may consist include at heat treatment at a
temperature of
about 82 C. for about 10 minutes. Alternatively, the enzyme can be
deactivated by
heating the solution to a temperature of about 92 C. for about 5 seconds.
After
enzyme deactivation is complete, the hydrolysate can be stored in a liquid
state at a
temperature lower than 10 C.
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[0099] In some embodiments, the protein equivalent source comprises a
hydrolyzed
protein, which includes partially hydrolyzed protein and extensively
hydrolyzed protein,
such as casein. In some embodiments, the protein equivalent source comprises a
hydrolyzed protein including peptides having a molar mass distribution of
greater than
500 Daltons. In some embodiments, the hydrolyzed protein comprises peptides
having
a molar mass distribution in the range of from about 500 Da!tons to about
1,500 Da!tons.
Still, in some embodiments the hydrolyzed protein may comprise peptides having
a
molar mass distribution range of from about 500 Da'tons to about 2,000
Da!tons.
[0100] In some embodiments, the protein equivalent source may comprise the
peptide
component, intact protein, hydrolyzed protein, including partially hydrolyzed
protein
and/or extensively hydrolyzed protein, and combinations thereof. In some
embodiments, 20% to 80% of the protein equivalent source comprises the peptide
component disclosed herein. In some embodiments, 30% to 60% of the protein
equivalent source comprises the peptide component disclosed herein. In still
other
embodiments, 40% to 50% of the protein equivalent source comprises the peptide
component.
[0101] In some embodiments, 20% to 80% of the protein equivalent source
comprises
intact protein, partially hydrolyzed protein, extensively hydrolyzed protein,
or
combinations thereof. In some embodiments, 40% to 70% of the protein
equivalent
source comprises intact proteins, partially hydrolyzed proteins, extensively
hydrolyzed
protein, or a combination thereof. In still further embodiments, 50% to 60% of
the
protein equivalent source may comprise intact proteins, partially hydrolyzed
protein,
extensively hydrolyzed protein, or a combination thereof.
[0102] In some embodiments the protein equivalent source comprises partially
hydrolyzed protein having a degree of hydrolysis of less than 40%. In still
other
embodiments, the protein equivalent source may comprise partially hydrolyzed
protein
having a degree of hydrolysis of less than 25%, or less than 15%.
[0103] In some embodiments, the nutritional composition comprises between
about 1 g
and about 7 g of a protein equivalent source per 100 Kcal. In other
embodiments, the
nutritional composition comprises between about 3.5 g and about 4.5 g of
protein
equivalent source per 100 Kcal.
[0104] Without being bound by any particular theory, the administration of a
nutritional
composition as disclosed herein may reduce allergic response and may improve
tolerance to cow's milk allergy in certain subjects. In some embodiments, the
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combination of probiotic, such as LGG, dietary butyrate, and the protein
equivalent
source provide synergistic health benefits.
[0105] The nutritional composition(s) of the present disclosure may also
comprise a
carbohydrate source. Carbohydrate sources can be any used in the art, e.g.,
lactose,
glucose, fructose, corn syrup solids, maltodextrins, sucrose, starch, rice
syrup solids, and
the like. The amount of carbohydrate in the nutritional composition typically
can vary
from between about 5 g and about 25 g/100 Kcal. In some embodiments, the
amount
of carbohydrate is between about 6 g and about 22 g/ 100 Kcal. In other
embodiments, the amount of carbohydrate is between about 12 g and about 14
g/100 Kcal. In some embodiments, corn syrup solids are preferred. Moreover,
hydrolyzed, partially hydrolyzed, and/or extensively hydrolyzed carbohydrates
may be
desirable for inclusion in the nutritional composition due to their easy
digestibility.
Specifically, hydrolyzed carbohydrates are less likely to contain allergenic
epitopes.
[0106] Non-limiting examples of carbohydrate materials suitable for use herein
include
hydrolyzed or intact, naturally or chemically modified, starches sourced from
corn,
tapioca, rice or potato, in waxy or non-waxy forms. Non-limiting examples of
suitable
carbohydrates include various hydrolyzed starches characterized as hydrolyzed
cornstarch, maltodextrin, maltose, corn syrup, dextrose, corn syrup solids,
glucose, and
various other glucose polymers and combinations thereof. Non-limiting examples
of
other suitable carbohydrates include those often referred to as sucrose,
lactose,
fructose, high fructose corn syrup, indigestible oligosaccharides such as
fructooligosaccharides and combinations thereof.
[0107] The nutritional composition(s) of the disclosure may also comprise a
protein
source. The protein source can be any used in the art, e.g., nonfat milk, whey
protein,
casein, soy protein, hydrolyzed protein, amino acids, and the like. Bovine
milk protein
sources useful in practicing the present disclosure include, but are not
limited to, milk
protein powders, milk protein concentrates, milk protein isolates, nonfat milk
solids,
nonfat milk, nonfat dry milk, whey protein, whey protein isolates, whey
protein
concentrates, sweet whey, acid whey, casein, acid casein, caseinate (e.g.
sodium
caseinate, sodium calcium caseinate, calcium caseinate) and any combinations
thereof.
[0108] In one embodiment, the proteins of the nutritional composition are
provided as
intact proteins. In other embodiments, the proteins are provided as a
combination of
both intact proteins and partially hydrolyzed proteins, with a degree of
hydrolysis of
between about 4% and 10%. In certain other embodiments, the proteins are more
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completely hydrolyzed. In still other embodiments, the protein source
comprises amino
acids. In yet another embodiment, the protein source may be supplemented with
glutamine-containing peptides.
[0109] In a particular embodiment of the nutritional composition, the
whey:casein ratio
of the protein source is similar to that found in human breast milk. In an
embodiment,
the protein source comprises from about 40% to about 80% whey protein and from
about 20% to about 60% casein.
[0110] In some embodiments, the nutritional composition comprises between
about 1 g
and about 7 g of a protein source per 100 Kcal. In other embodiments, the
nutritional
composition comprises between about 3.5 g and about 4.5 g of protein per 100
Kcal.
[0111] In some embodiments, the nutritional composition described herein
comprises a
fat source. The enriched lipid fraction described herein may be the sole fat
source or
may be used in combination with any other suitable fat or lipid source for the
nutritional
composition as known in the art. In certain embodiments, appropriate fat
sources
include, but are not limited to, animal sources, e.g., milk fat, butter,
butter fat, egg yolk
lipid; marine sources, such as fish oils, marine oils, single cell oils;
vegetable and plant
oils, such as corn oil, canola oil, sunflower oil, soybean oil, palm olein
oil, coconut oil,
high oleic sunflower oil, evening primrose oil, rapeseed oil, olive oil,
flaxseed (linseed)
oil, cottonseed oil, high oleic safflower oil, palm stearin, palm kernel oil,
wheat germ oil;
medium chain triglyceride oils and emulsions and esters of fatty acids; and
any
combinations thereof.
[0112] In some embodiment the nutritional composition comprises between about
1
g/100 Kcal to about 10 g/100 Kcal of a fat or lipid source. In some
embodiments, the
nutritional composition comprises between about 2 g/100 Kcal to about 7 g/100
Kcal
of a fat source. In other embodiments the fat source may be present in an
amount
from about 2.5 g/100 Kcal to about 6 g/100 Kcal. In still other embodiments,
the fat
source may be present in the nutritional composition in an amount from about 3
g/100
Kcal to about 4 g/100 Kcal.
[0113] In some embodiments, the fat or lipid source comprises from about 10%
to about
35% palm oil per the total amount of fat or lipid. In some embodiments, the
fat or lipid
source comprises from about 15% to about 30% palm oil per the total amount of
fat or
lipid. Yet in other embodiments, the fat or lipid source may comprise from
about 18% to
about 25 % palm oil per the total amount of fat or lipid.
[0114] In certain embodiments, the fat or lipid source may be formulated to
include
from about 2% to about 16% soybean oil based on the total amount of fat or
lipid. In
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some embodiments, the fat or lipid source may be formulated to include from
about
4% to about 12% soybean oil based on the total amount of fat or lipid. In some
embodiments, the fat or lipid source may be formulated to include from about
6% to
about 10% soybean oil based on the total amount of fat or lipid.
[0115] In certain embodiments, the fat or lipid source may be formulated to
include
from about 2% to about 16% coconut oil based on the total amount of fat or
lipid. In
some embodiments, the fat or lipid source may be formulated to include from
about
4% to about 12% coconut oil based on the total amount of fat or lipid. In some
embodiments, the fat or lipid source may be formulated to include from about
6% to
about 10% coconut oil based on the total amount of fat or lipid.
[0116] In certain embodiments, the fat or lipid source may be formulated to
include
from about 2% to about 16% sunflower oil based on the total amount of fat or
lipid. In
some embodiments, the fat or lipid source may be formulated to include from
about
4% to about 12% sunflower oil based on the total amount of fat or lipid. In
some
embodiments, the fat or lipid source may be formulated to include from about
6% to
about 10% sunflower oil based on the total amount of fat or lipid.
[0117] In some embodiments, the oils, i.e. sunflower oil, soybean oil,
sunflower oil, palm
oil, etc. are meant to cover fortified versions of such oils known in the art.
For example,
in certain embodiments, the use of sunflower oil may include high oleic
sunflower oil. In
other examples, the use of such oils may be fortified with certain fatty
acids, as known
in the art, and may be used in the fat or lipid source disclosed herein.
[0118] In some embodiments the nutritional composition may also include a
source of
LCPUFAs. In one embodiment the amount of LCPUFA in the nutritional composition
is
advantageously at least about 5 mg/100 Kcal, and may vary from about 5 mg/100
Kcal to about 100 mg/100 Kcal, more preferably from about 10 mg/100 Kcal to
about
50 mg/100 Kcal. Non-limiting examples of LCPUFAs include, but are not limited
to, DHA,
ARA, linoleic (18:2 n-6), y-linolenic (18:3 n-6), dihomo- y-linolenic (20:3 n-
6) acids in the
n-6 pathway, a-linolenic (18:3 n-3), stearidonic (18:4 n-3), eicosatetraenoic
(20:4 n-3),
eicosapentaenoic (20:5 n-3), and docosapentaenoic (22:6 n-3).
[0119] In some embodiments, the LCPUFA included in the nutritional composition
may
comprise DHA. In one embodiment the amount of DHA in the nutritional
composition is
advantageously at least about 17 mg/100 Kcal, and may vary from about 5 mg/100
Kcal to about 75 mg/100 Kcal, more preferably from about 10 mg/100 Kcal to
about 50
mg/100 Kcal.
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[0120] In another embodiment, especially if the nutritional composition is an
infant
formula, the nutritional composition is supplemented with both DHA and ARA. In
this
embodiment, the weight ratio of ARA:DHA may be between about 1:3 and about
9:1.
In a particular embodiment, the ratio of ARA:DHA is from about 1:2 to about
4:1.
[0121] The DHA and ARA can be in natural form, provided that the remainder of
the
LCPUFA source does not result in any substantial deleterious effect on the
infant.
Alternatively, the DHA and ARA can be used in refined form.
[0122] The disclosed nutritional composition described herein can, in some
embodiments, also comprise a source of B-glucan. Glucans are polysaccharides,
specifically polymers of glucose, which are naturally occurring and may be
found in
cell walls of bacteria, yeast, fungi, and plants. Beta glucans (13-glucans)
are themselves
a diverse subset of glucose polymers, which are made up of chains of glucose
monomers linked together via beta-type glycosidic bonds to form complex
carbohydrates.
[0123][3-1,3-glucans are carbohydrate polymers purified from, for example,
yeast,
mushroom, bacteria, algae, or cereals. (Stone BA, Clarke AE. Chemistry and
Biology of
(1-3)-Beta-Glucans. London:Portland Press Ltd; 1993.) The chemical structure
of 13-1,3-
glucan depends on the source of the P-1,3-glucan. Moreover, various
physiochemical
parameters, such as solubility, primary structure, molecular weight, and
branching, play
a role in biological activities of P-1,3-glucans. (Yadomae T., Structure and
biological
activities of fungal beta- 1,3-glucans. Yakugaku Zasshi. 2000;120:413-431.)
[0124] 3-1,3-glucans are naturally occurring polysaccharides, with or without
13-1,6-
glucose side chains that are found in the cell walls of a variety of plants,
yeasts, fungi
and bacteria. 3-1,3;1,6-glucans are those containing glucose units with (1,3)
links
having side chains attached at the (1,6) position(s). 13-1,3;1,6 glucans are a
heterogeneous group of glucose polymers that share structural commonalities,
including a backbone of straight chain glucose units linked by a 13-1,3 bond
withP-1,6-
linked glucose branches extending from this backbone. While this is the basic
structure
for the presently described class of P-glucans, some variations may exist. For
example,
certain yeast 13-glucans have additional regions of P(1,3) branching extending
from the
13(1,6) branches, which add further complexity to their respective structures.
[0125] 13-glucans derived from baker's yeast, Saccharomyces cerevisiae, are
made up
of chains of D-glucose molecules connected at the 1 and 3 positions, having
side
chains of glucose attached at the 1 and 6 positions. Yeast-derived P-glucan is
an
insoluble, fiber-like, complex sugar having the general structure of a linear
chain of
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glucose units with a 13-1,3 backbone interspersed with 13-1,6 side chains that
are
generally 6-8 glucose units in length. More specifically, 13-glucan derived
from baker's
yeast is poly-(1,6)-13-D-glucopyranosyl-(1,3)-13-D-glucopyranose.
[0126] Furthermore, 13-glucans are well tolerated and do not produce or cause
excess
gas, abdominal distension, bloating or diarrhea in pediatric subjects.
Addition of 13-
glucan to a nutritional composition for a pediatric subject, such as an infant
formula, a
growing-up milk or another children's nutritional product, will improve the
subject's
immune response by increasing resistance against invading pathogens and
therefore
maintaining or improving overall health.
[0127] In some embodiments, the 13-glucan is 13-1,3;1,6-glucan. In some
embodiments,
the 13-1,3;1,6-glucan is derived from baker's yeast. The nutritional
composition may
comprise whole glucan particle 13-glucan, particulate 13-glucan, PGG-glucan
(poly-1,6-
13-D-glucopyranosy1-1,3-13-D-glucopyranose) or any mixture thereof.
[0128] In some embodiments, the amount of 13-glucan in the nutritional
composition is
between about 3 mg and about 17 mg per 100 Kcal. In another embodiment the
amount of (3-glucan is between about 6 mg and about 17 mg per 100 Kcal.
[0129] The disclosed nutritional composition described herein, can, in some
embodiments also comprise an effective amount of iron. The iron may comprise
encapsulated iron forms, such as encapsulated ferrous fumarate or encapsulated
ferrous sulfate or less reactive iron forms, such as ferric pyrophosphate or
ferric
orthophosphate.
[0130] One or more vitamins and/or minerals may also be added in to the
nutritional
composition in amounts sufficient to supply the daily nutritional requirements
of a
subject. It is to be understood by one of ordinary skill in the art that
vitamin and mineral
requirements will vary, for example, based on the age of the child. For
instance, an
infant may have different vitamin and mineral requirements than a child
between the
ages of one and thirteen years. Thus, the embodiments are not intended to
limit the
nutritional composition to a particular age group but, rather, to provide a
range of
acceptable vitamin and mineral components.
[0131] In embodiments providing a nutritional composition for a child, the
composition
may optionally include, but is not limited to, one or more of the following
vitamins or
derivations thereof: vitamin 131 (thiamin, thiamin pyrophosphate, TPP, thiamin
triphosphate, TTP, thiamin hydrochloride, thiamin mononitrate), vitamin B2
(riboflavin,
flavin mononucleotide, FMN, flavin adenine dinucleotide, FAD, lactoflavin,
ovoflavin),
vitamin B3 (niacin, nicotinic acid, nicotinamide, niacinamide, nicotinamide
adenine
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dinucleotide, NAD, nicotinic acid mononucleotide, NicMN, pyridine-3-carboxylic
acid),
vitamin B3-precursor tryptophan, vitamin B6 (pyridoxine, pyridoxal,
pyridoxamine,
pyridoxine hydrochloride), pantothenic acid (pantothenate, panthenol), folate
(folic
acid, folacin, pteroylglutamic acid), vitamin B12 (cobalamin, methylcobalamin,
deoxyadenosylcobalamin, cyanocobalamin, hydroxycobalamin, adenosylcobalamin),
biotin, vitamin C (ascorbic acid), vitamin A (retinol, retinyl acetate,
retinyl palmitate,
retinyl esters with other long-chain fatty acids, retinal, retinoic acid,
retinol esters),
vitamin D (calciferol, cholecalciferol, vitamin D3, 1,25,-dihydroxyvitamin D),
vitamin E (a-
tocopherol, a-tocopherol acetate, a-tocopherol succinate, a-tocopherol
nicotinate,
a-tocopherol), vitamin K (vitamin Ki, phylloquinone, naphthoquinone, vitamin
K2,
menaquinone-7, vitamin K3, menaquinone-4, menadione, menaquinone-8,
menaquinone-8H, menaquinone-9, menaquinone-9H, menaquinone-10,
menaquinone-11, menaquinone-12, menaquinone-13), chorine, inosito1,13-carotene
and any combinations thereof.
[0132] In embodiments providing a children's nutritional product, such as a
growing-up
milk, the composition may optionally include, but is not limited to, one or
more of the
following minerals or derivations thereof: boron, calcium, calcium acetate,
calcium
gluconate, calcium chloride, calcium lactate, calcium phosphate, calcium
sulfate,
chloride, chromium, chromium chloride, chromium picolonate, copper, copper
sulfate,
copper gluconate, cupric sulfate, fluoride, iron, carbonyl iron, ferric iron,
ferrous
fumarate, ferric orthophosphate, iron trituration, polysaccharide iron,
iodide, iodine,
magnesium, magnesium carbonate, magnesium hydroxide, magnesium oxide,
magnesium stearate, magnesium sulfate, manganese, molybdenum, phosphorus,
potassium, potassium phosphate, potassium iodide, potassium chloride,
potassium
acetate, selenium, sulfur, sodium, docusate sodium, sodium chloride, sodium
selenate,
sodium molybdate, zinc, zinc oxide, zinc sulfate and mixtures thereof. Non-
limiting
exemplary derivatives of mineral compounds include salts, alkaline salts,
esters and
cherates of any mineral compound.
[0133] The minerals can be added to growing-up milks or to other children's
nutritional
compositions in the form of salts such as calcium phosphate, calcium glycerol
phosphate, sodium citrate, potassium chloride, potassium phosphate, magnesium
phosphate, ferrous sulfate, zinc sulfate, cupric sulfate, manganese sulfate,
and sodium
selenite. Additional vitamins and minerals can be added as known within the
art.
[0134] The nutritional compositions of the present disclosure may optionally
include one
or more of the following flavoring agents, including, but not limited to,
flavored
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extracts, volatile oils, cocoa or chocolate flavorings, peanut butter
flavoring, cookie
crumbs, vanilla or any commercially available flavoring. Examples of useful
flavorings
include, but are not limited to, pure anise extract, imitation banana extract,
imitation
cherry extract, chocolate extract, pure lemon extract, pure orange extract,
pure
peppermint extract, honey, imitation pineapple extract, imitation rum extract,
imitation
strawberry extract, or vanilla extract; or volatile oils, such as balm oil,
bay oil, bergamot
oil, cedarwood oil, cherry oil, cinnamon oil, clove oil, or peppermint oil;
peanut butter,
chocolate flavoring, vanilla cookie crumb, butterscotch, toffee, and mixtures
thereof.
The amounts of flavoring agent can vary greatly depending upon the flavoring
agent
used. The type and amount of flavoring agent can be selected as is known in
the art.
[0135] The nutritional compositions of the present disclosure may optionally
include one
or more emulsifiers that may be added for stability of the final product.
Examples of
suitable emulsifiers include, but are not limited to, lecithin (e.g., from egg
or soy), alpha
lactalbumin and/or mono- and di-glycerides, and mixtures thereof. Other
emulsifiers
are readily apparent to the skilled artisan and selection of suitable
emulsifier(s) will
depend, in part, upon the formulation and final product.
[0136] The nutritional compositions of the present disclosure may optionally
include one
or more preservatives that may also be added to extend product shelf life.
Suitable
preservatives include, but are not limited to, potassium sorbate, sodium
sorbate,
potassium benzoate, sodium benzoate, calcium disodium EDTA, and mixtures
thereof.
[0137] The nutritional compositions of the present disclosure may optionally
include one
or more stabilizers. Suitable stabilizers for use in practicing the
nutritional composition of
the present disclosure include, but are not limited to, gum arabic, gum
ghatti, gum
karaya, gum tragacanth, agar, furcellaran, guar gum, gellan gum, locust bean
gum,
pectin, low methoxyl pectin, gelatin, microcrystalline cellulose, CMC (sodium
carboxymethylcellulose), methylcellulose hydroxypropyl methyl cellulose,
hydroxypropyl cellulose, DATEM (diacetyl tartaric acid esters of mono- and
diglycerides), dextran, carrageenans, and mixtures thereof.
[0138] The nutritional compositions of the disclosure may provide minimal,
partial or
total nutritional support. The compositions may be nutritional supplements or
meal
replacements. The compositions may, but need not, be nutritionally complete.
In an
embodiment, the nutritional composition of the disclosure is nutritionally
complete and
contains suitable types and amounts of lipid, carbohydrate, protein, vitamins
and
minerals. The amount of lipid or fat typically can vary from about 1 to about
25 g/100
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Kcal. The amount of protein typically can vary from about 1 to about 7 g/100
Kcal. The
amount of carbohydrate typically can vary from about 6 to about 22 g/100 Kcal.
[0139] In an embodiment, the children's nutritional composition may contain
between
about 10 and about 50% of the maximum dietary recommendation for any given
country, or between about 10 and about 50% of the average dietary
recommendation
for a group of countries, per serving of vitamins A, C, and E, zinc, iron,
iodine, selenium,
and choline. In another embodiment, the children's nutritional composition may
supply about 10 - 30% of the maximum dietary recommendation for any given
country,
or about 10 - 30% of the average dietary recommendation for a group of
countries,
per serving of B-vitamins. In yet another embodiment, the levels of vitamin D,
calcium,
magnesium, phosphorus, and potassium in the children's nutritional product may
correspond with the average levels found in milk. In other embodiments, other
nutrients in the children's nutritional composition may be present at about
20% of the
maximum dietary recommendation for any given country, or about 20% of the
average dietary recommendation for a group of countries, per serving.
[0140] In some embodiments the nutritional composition is an infant formula.
Infant
formulas are fortified nutritional compositions for an infant. The content of
an infant
formula is dictated by federal regulations, which define macronutrient,
vitamin,
mineral, and other ingredient levels in an effort to simulate the nutritional
and other
properties of human breast milk. Infant formulas are designed to support
overall health
and development in a pediatric human subject, such as an infant or a child.
[0141] In some embodiments, the nutritional composition of the present
disclosure is a
growing-up milk. Growing-up milks are fortified milk-based beverages intended
for
children over 1 year of age (typically from 1-3 years of age, from 4-6 years
of age or
from 1-6 years of age). They are not medical foods and are not intended as a
meal
replacement or a supplement to address a particular nutritional deficiency.
Instead,
growing-up milks are designed with the intent to serve as a complement to a
diverse
diet to provide additional insurance that a child achieves continual, daily
intake of all
essential vitamins and minerals, macronutrients plus additional functional
dietary
components, such as non-essential nutrients that have purported health-
promoting
properties.
[0142] The exact composition of a growing-up milk or other nutritional
composition
according to the present disclosure can vary from market-to-market, depending
on
local regulations and dietary intake information of the population of
interest. In some
embodiments, nutritional compositions according to the disclosure consist of a
milk
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protein source, such as whole or skim milk, plus added sugar and sweeteners to
achieve desired sensory properties, and added vitamins and minerals. The fat
composition includes an enriched lipid fraction derived from milk. Total
protein can be
targeted to match that of human milk, cow milk or a lower value. Total
carbohydrate is
usually targeted to provide as little added sugar, such as sucrose or
fructose, as
possible to achieve an acceptable taste. Typically, Vitamin A, calcium and
Vitamin D
are added at levels to match the nutrient contribution of regional cow milk.
Otherwise,
in some embodiments, vitamins and minerals can be added at levels that provide
approximately 20% of the dietary reference intake (DRI) or 20% of the Daily
Value (DV)
per serving. Moreover, nutrient values can vary between markets depending on
the
identified nutritional needs of the intended population, raw material
contributions and
regional regulations.
[0143] The disclosed nutritional composition(s) may be provided in any form
known in
the art, such as a powder, a gel, a suspension, a paste, a solid, a liquid, a
liquid
concentrate, a reconstituteable powdered milk substitute or a ready-to-use
product.
The nutritional composition may, in certain embodiments, comprise a
nutritional
supplement, children's nutritional product, infant formula, human milk
fortifier, growing-
up milk or any other nutritional composition designed for an infant or a
pediatric
subject. Nutritional compositions of the present disclosure include, for
example, orally-
ingestible, health-promoting substances including, for example, foods,
beverages,
tablets, capsules and powders. Moreover, the nutritional composition of the
present
disclosure may be standardized to a specific caloric content, it may be
provided as a
ready-to-use product, or it may be provided in a concentrated form. In some
embodiments, the nutritional composition is in powder form with a particle
size in the
range of 5 pm to 1500 pm, more preferably in the range of 10 pm to 300pm.
[0144] The nutritional compositions of the present disclosure may be provided
in a
suitable container system. For example, non-limiting examples of suitable
container
systems include plastic containers, metal containers, foil pouches, plastic
pouches,
multi-layered pouches, and combinations thereof. In certain embodiments, the
nutritional composition may be a powdered composition that is contained within
a
plastic container. In certain other embodiments, the nutritional composition
may be
contained within a plastic pouch located inside a plastic container.
[0145] The nutritional compositions described herein, in some embodiments,
advantageously reduce the incidence of allergic reaction and improve tolerance
to
cow's milk allergy in a subject. Further, in some embodiments, the nutritional
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compositions advantageously reduce the inflammatory response caused by allergy
in
a subject. Accordingly, the disclosure relates to methods of improving
tolerance to
cow's milk allergy in a subject. Further, the disclosure relates to methods
for the dietary
management of allergic diseases and/or allergic reaction in a subject via
administration of the nutritional compositions including a probiotic, such as
LGG, and
dietary butyrate as disclosed herein.
[0146] In some embodiments, the method comprises the step of subjecting the
target
subject to cow's milk and then providing the nutritional composition disclosed
herein
including a probiotic and dietary butyrate to the target subject. In certain
embodiments, after the target subject has been subjected to cow's milk, the
target
subject may be provided with a nutritional composition that includes a
probiotic and
dietary butyrate and a protein equivalent source as disclosed herein. In
certain
embodiments, the target subject, after being exposed to cow's milk or other
allergen,
may be administered a nutritional composition comprising probiotic and dietary
butyrate, and a protein equivalent source. In certain embodiments, the protein
equivalent source may be substantially free of whole and/or intact protein. In
certain
other embodiments, the protein equivalent source may comprise hydrolyzed
protein,
amino acids, the peptide component disclosed herein, and combinations thereof.
In
some embodiments, the nutritional composition includes a protein equivalent
source
includes amino acids and no hydrolyzed or whole/intact protein.
[0147] In some embodiments, the target subject is not subjected to cow's milk
or an
allergen prior to administration of the nutritional composition. Thus, in some
embodiments, the method is directed to reducing allergic response in a target
subject
via providing the nutritional compositions disclosed herein including dietary
butyrate to
the target subject, and subsequently exposing the target subject to cow's milk
or other
allergen.
[0148] The nutritional compositions described herein, in some embodiments,
advantageously reduce the inflammatory response in a subject. Accordingly, the
disclosure relates to methods of reducing a proinflammatory response in a
subject by
administering to a subject a nutritional composition containing the protein
equivalent
source described herein in combination with probiotic and dietary butyrate.
For
example, the present methods may reduce the production of proinflammatory
cytokines in a subject.
[0149] In some embodiments, the method for reducing an inflammatory response
in a
subject comprises administering to a subject a nutritional composition
comprising a
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carbohydrate source, a protein equivalent source, fat source, probiotic, and
dietary
butyrate, wherein the protein equivalent source includes a peptide component
comprising SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO
24, SEQ
ID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO
60, and
SEQ ID NO 63. In some embodiments, the peptide component may comprise
additional peptides disclosed in Table 1. For example, the composition may
include at
least 10 additional peptides disclosed in Table 1. In some embodiments, 20% to
80% of
the protein equivalent source comprises the peptide component, and 20% to 80%
of
the protein equivalent source comprises an intact protein, a partially
hydrolyzed
protein, and combinations thereof.
[0150] In another embodiment, the method comprises administering to a subject
a
nutritional composition, wherein 20% to 80% of the protein equivalent source
includes a
peptide component comprising at least 3 peptides selected from the group
consisting
of SEQ ID NO 4, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 21, SEQ ID NO 24, SEQ ID
NO 30,
SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 51, SEQ ID NO 57, SEQ ID NO 60, and SEQ
ID NO
63, and at least 5 additional peptides selected from Table 1; and wherein 20%
to 80% of
the protein equivalent source comprises an intact protein, a partially
hydrolyzed
protein, or combinations thereof.
[0151] In yet other embodiments, the method for reducing the inflammatory
response
includes providing a nutritional composition comprising a peptide component
from
Table 1, wherein the peptide component is derived from a casein hydrolysate
having a
molar mass distribution of greater than 500 Daltons. In some embodiments, the
molar
mass distribution of the casein hydrolysate is in a range of 500 to 2000
Daltons. In other
embodiments, the method for reducing the inflammatory response includes
providing
a nutritional composition comprising the peptide component described herein,
wherein the peptide component is derived from a casein hydrolysate that does
not
include peptides having a molar mass distribution of less than 200 Daltons.
[0152] In some embodiments the target subject may be a pediatric subject.
Further, in
one embodiment, the nutritional composition provided to the pediatric subject
may be
an infant formula. The peptide component identified herein, probiotic and
dietary
butyrate as disclosed herein may be added to the infant formula and, further,
each
may be selected from a specific source and concentrations thereof may be
adjusted
to maximize health benefits. In another embodiment of this method, the
nutritional
composition comprising the peptide component disclosed herein, probiotic and
dietary butyrate is a growing up milk.
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[0153] In embodiments when the nutritional composition is an infant formula,
the
composition may advantageously reduce a pro-inflammatory response in the
infant,
and thereby reduce the incidence of inflammatory disease. Moreover, the
reduction
in inflammatory disease may last throughout childhood and into adulthood.
Similarly,
when the nutritional composition is a growing-up milk, a child who ingests the
growing-
up milk may experience a reduction in the incidence of inflammatory disease in
adulthood, as well as during childhood.
[0154] In certain embodiments, the disclosure is directed to a method for
improving the
absorption of butyrate in a target subject by providing or administering the
nutritional
compositions disclosed herein including probiotic and dietary butyrate to the
target
subject. In some embodiments, the target subject is a pediatric subject or an
infant. In
some embodiments, the nutritional composition is an infant formula or a
growing-up
milk.
[0155] All combinations of method or process steps as used herein can be
performed in
any order, unless otherwise specified or clearly implied to the contrary by
the context in
which the referenced combination is made.
[0156] The methods and compositions of the present disclosure, including
components
thereof, can comprise, consist of, or consist essentially of the essential
elements and
limitations of the embodiments described herein, as well as any additional or
optional
ingredients, components or limitations described herein or otherwise useful in
nutritional
compositions.
[0157] Formulation examples are provided to illustrate some embodiments of the
nutritional composition of the present disclosure but should not be
interpreted as any
limitation thereon. Other embodiments within the scope of the claims herein
will be
apparent to one skilled in the art from the consideration of the specification
or practice
of the nutritional composition or methods disclosed herein. It is intended
that the
specification, together with the example, be considered to be exemplary only,
with the
scope and spirit of the disclosure being indicated by the claims which follow
the
example.
FORMULATION EXAMPLES
[0158] Table 3 provides an example embodiment of a peptide component including
8
peptides from Table1.
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Table 3. Example peptide component
Example of Selected Peptides
for Peptide Component
SEQ ID NO 5
SEQ ID NO 24
SEQ ID NO 33
SEQ ID NO 56
SEQ ID NO 64
SEQ ID NO 13
SEQ ID NO 24
SEQ ID NO 60
[0159] Table 4 provides an example embodiment of a peptide component including
certain peptides from Table 1.
Table 4. Example peptide component
Example of Selected Peptides
for Peptide Component
SEQ ID NO 13
SEQ ID NO 24
SEQ ID NO 60
SEQ ID NO 5
SEQ ID NO 11
SEQ ID NO 22
SEQ ID NO 25
SEQ ID NO 33
SEQ ID NO 45
SEQ ID NO 46
SEQ ID NO 47
SEQ ID NO 48
SEQ ID NO 52
SEQ ID NO 34
SEQ ID NO 36
SEQ ID NO 61
SEQ ID NO 62
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SEQ ID NO 64
Table 5
[0160] Table 5, illustrated below, provides an example embodiment of the
nutritional
profile of a nutritional composition including PDX/GOS and dietary butyrate
and
describes the amount of each ingredient to be included per 100 Kcal serving of
nutritional composition.
Table 5. Nutrition profile of an example nutritional composition including
dietary
butyrate
per 100 Kcal
Nutrient
Minimum Maximum
Protein Equivalent Source (g) 1.0 7.0
Dietary butyrate (mg) 22 280
Lactobacillus rhamnosus GG (cfu) 1x104 1.5x1012
Carbohydrates (g) 6 22
Fat (g) 1.3 7.2
Pre biotic (g) 0.3 1.2
DHA (g) 4 22
Beta glucan (mg) 2.9 17
Probiotics (cfu) 0.5 5.0
Vitamin A (1U) 9.60x 108 3.80 x 108
Vitamin D (1U) 134 921
Vitamin E (1U) 22 126
Vitamin K (mcg) 0.8 5.4
Thiamin (mcg) 2.9 18
Riboflavin (mcg) 63 328
Vitamin B6 (mcg) 68 420
Vitamin B12 (mcg) 52 397
Niacin (mcg) 0.2 0.9
Folic acid (mcg) 690 5881
Panthothenic acid (mcg) 8 66
Biotin (mcg) 232 1 211
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Vitamin C (mg) 1.4 5.5
Choline (mg) 4.9 24
Calcium (mg) 4.9 43
Phosphorus (mg) 68 297
Magnesium (mg) 54 210
Sodium (mg) 4.9 34
Potassium (mg) 24 88
Chloride (mg) 82 346
Iodine (mcg) 53 237
Iron (mg) 8.9 79
Zinc (mg) 0.7 2.8
Manganese (mcg) 0.7 2.4
Copper (mcg) 7.2 41
[0161] All references cited in this specification, including without
limitation, all papers,
publications, patents, patent applications, presentations, texts, reports,
manuscripts,
brochures, books, internet postings, journal articles, periodicals, and the
like, are hereby
incorporated by reference into this specification in their entireties. The
discussion of the
references herein is intended merely to summarize the assertions made by their
authors
and no admission is made that any reference constitutes prior art. Applicants
reserve
the right to challenge the accuracy and pertinence of the cited references.
[0162] Although embodiments of the disclosure have been described using
specific
terms, devices, and methods, such description is for illustrative purposes
only. The
words used are words of description rather than of limitation. It is to be
understood that
changes and variations may be made by those of ordinary skill in the art
without
departing from the spirit or the scope of the present disclosure, which is set
forth in the
following claims. In addition, it should be understood that aspects of the
various
embodiments may be interchanged in whole or in part. Therefore, the spirit and
scope
of the appended claims should not be limited to the description of the
versions
contained therein.
