U.S. patent application number 13/605223 was filed with the patent office on 2013-03-07 for nutritional compositions containing human milk oligosaccharides and methods for using the same.
This patent application is currently assigned to Prolacta Bioscience, Inc.. The applicant listed for this patent is Scott Eaker, Scott Elster, Joseph FOURNELL, David J. Rechtman. Invention is credited to Scott Eaker, Scott Elster, Joseph FOURNELL, David J. Rechtman.
Application Number | 20130059815 13/605223 |
Document ID | / |
Family ID | 42233595 |
Filed Date | 2013-03-07 |
United States Patent
Application |
20130059815 |
Kind Code |
A1 |
FOURNELL; Joseph ; et
al. |
March 7, 2013 |
NUTRITIONAL COMPOSITIONS CONTAINING HUMAN MILK OLIGOSACCHARIDES AND
METHODS FOR USING THE SAME
Abstract
This disclosure features human milk permeates and compositions
containing the same obtained from fractionated whole human milk.
The oligosaccharide rich permeate and permeate compositions of the
present invention are useful as nutritional supplements for
pre-term and full term infants, for establishing or maintaining gut
flora and for treating the symptoms of inflammatory bowel
disease.
Inventors: |
FOURNELL; Joseph; (Newbury
Park, CA) ; Eaker; Scott; (Thousand Oaks, CA)
; Elster; Scott; (Monrovia, CA) ; Rechtman; David
J.; (Hermosa Beach, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
FOURNELL; Joseph
Eaker; Scott
Elster; Scott
Rechtman; David J. |
Newbury Park
Thousand Oaks
Monrovia
Hermosa Beach |
CA
CA
CA
CA |
US
US
US
US |
|
|
Assignee: |
Prolacta Bioscience, Inc.
Monrovia
CA
|
Family ID: |
42233595 |
Appl. No.: |
13/605223 |
Filed: |
September 6, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13132545 |
Jul 7, 2011 |
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PCT/US2009/066430 |
Dec 2, 2009 |
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13605223 |
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61119176 |
Dec 2, 2008 |
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61193546 |
Dec 5, 2008 |
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Current U.S.
Class: |
514/54 ; 426/2;
426/580; 426/61; 426/72 |
Current CPC
Class: |
A23C 9/1422 20130101;
A23L 33/40 20160801; A61K 35/20 20130101; A61P 1/00 20180101; A61P
1/04 20180101; A23C 9/148 20130101; A23V 2002/00 20130101; A23C
9/1522 20130101; A61P 29/00 20180101; A23C 3/02 20130101; A23C
9/206 20130101; A23C 9/1427 20130101; A23C 9/158 20130101; A61P
3/02 20180101; A23V 2002/00 20130101; A23V 2250/28 20130101; A23V
2200/3202 20130101 |
Class at
Publication: |
514/54 ; 426/580;
426/72; 426/61; 426/2 |
International
Class: |
A23C 9/152 20060101
A23C009/152; A61P 1/04 20060101 A61P001/04; A61K 31/702 20060101
A61K031/702; A61P 1/00 20060101 A61P001/00; A23C 9/00 20060101
A23C009/00; A23C 9/158 20060101 A23C009/158 |
Claims
1-63. (canceled)
64. A nutritional supplement comprising human milk
oligosaccharides.
65. The nutritional supplement of claim 64, wherein said
nutritional supplement is administered as a part of a non-human
milk baby formula.
66. The nutritional supplement of claim 64 wherein said nutritional
supplement comprises neutral and/or anionic human milk
oligosaccharides.
67. The nutritional supplement of claim 66 wherein the neutral
and/or anionic human milk oligosaccharides contain galactose,
N-acetylglucosamine, lactose and/or fucose.
68. The nutritional supplement of claim 64 wherein said human milk
oligosaccharides are fucosilated and/or sialilated.
69. The nutritional supplement of claim 64 further comprising
vitamins and/or minerals.
70. The nutritional supplement of claim 64 further comprising a
probiotic.
71. The nutritional supplement of claim 70 wherein the probiotic is
a lactobacilli and/or a bifidobacteria.
72. The nutritional supplement of claim 71 wherein the probiotic is
a bifidobacteria.
73. A method for promoting the development of beneficial gut flora
in preterm infants or neonates or maintaining proper gut flora in
children and adults comprising administering a nutritional
supplement comprising human milk oligosaccharides.
74. The method of claim 73 wherein said nutritional supplement is
added to a human milk formulation or a non-human milk formulation
prior to administration.
75. The method of claim 74, wherein said nutritional supplement is
added to a non-human milk formulation prior to administration.
76. The method of claim 73 wherein said nutritional supplement
comprises neutral and/or anionic human milk oligosaccharides.
77. The method of claim 76 wherein the neutral and/or anionic human
milk oligosaccharides contain galactose, N-acetylglucosamine,
lactose and/or fucose.
78. The method of claim 73 wherein said human milk oligosaccharides
are fucosilated and/or sialilated.
79. The method of claim 73 further comprising vitamins and/or
minerals.
80. The method of claim 73 further comprising a probiotic.
81. The method of claim 80 wherein the probiotic is a lactobacilli
and/or a bifidobacteria.
82. The method of claim 81 wherein the probiotic is a
bifidobacteria.
83. A method for treating an inflammatory bowel disease comprising
administering a nutritional supplement comprising human milk
oligosaccharides.
84. The method of claim 83 wherein the inflammatory bowel disease
is selected from the group consisting of ulcerative colitis,
indeterminate colitis, microscopic colitis, collagenous colitis,
Crohn's disease, and irritable bowel disease.
85. The method of claim 83 wherein the nutritional supplement is
administered to a preterm infant, infant, child or adult.
86. The method of claim 83 wherein said nutritional supplement is
added to a human milk formulation or a non-human milk formulation
prior to administration.
87. The method of claim 86, wherein said nutritional supplement is
added to a non-human milk formulation prior to administration.
88. The method of claim 87 wherein said nutritional supplement
comprises neutral and/or anionic human milk oligosaccharides.
89. The method of claim 88 wherein the neutral and/or anionic human
milk oligosaccharides contain galactose, N-acetylglucosamine,
lactose and/or fucose.
90. The nutritional supplement of claim 83 wherein said human milk
oligosaccharides are fucosilated and/or sialilated.
91. A method of treating or preventing necrotizing Enterocolitis
comprising administering a nutritional composition comprising human
milk oligosaccharides.
92. The method of claim 91 wherein said nutritional supplement is
added to a human milk formulation or a non-human milk formulation
prior to administration.
93. The method of claim 92, wherein said nutritional supplement is
added to a non-human milk formulation prior to administration.
94. The method of claim 91 wherein said nutritional supplement
comprises neutral and/or anionic human milk oligosaccharides.
95. The method of claim 94 wherein the neutral and/or anionic human
milk oligosaccharides contain galactose, N-acetylglucosamine,
lactose and/or fucose.
96. The method of claim 91 wherein said human milk oligosaccharides
are fucosilated and/or sialilated.
97. The nutritional supplement of claim 64, wherein the human milk
oligosaccharides are derived from human milk.
98. The method of claim 73, wherein the human milk oligosaccharides
are derived from human milk.
99. The method of claim 83, wherein the human milk oligosaccharides
are derived from human milk.
100. The method of claim 91, wherein the human milk
oligosaccharides are derived from human milk.
Description
[0001] This application claims priority to U.S. Provisional
Application No. 61/193,546, filed Dec. 5, 2008 and U.S. Provisional
Application No. 61/119,176, filed Dec. 2, 2008, both of which are
incorporated herein in their entirety.
TECHNICAL FIELD
[0002] This disclosure is related to human milk permeate
compositions, e.g., compositions comprising human milk
oligosaccharides, and methods of making and using such
compositions.
BACKGROUND
[0003] Human milk is generally the food of choice for preterm and
full term infants because of its nutritional composition and
immunologic benefits. The source of human milk can be, e.g., a
donor or the infant's mother. The nutritional value of raw or
conventionally-processed donor milk, however, varies and, in most
instances, is not sufficient to meet the needs of preterm infants.
In addition, a possibility of bacterial, viral and other
contamination of raw donor milk exists. Even the mother's own milk
is often not nutritionally sufficient for the premature infant. It
is often desirable to feed preterm infants milk that is fortified
with various supplements, e.g., oligosaccharides. Compositions that
contain human milk oligosaccharides can also be useful as
supplements for infants, children, and adults who have or are at
risk of developing various disorders and diseases, e.g., infections
or immune deficiencies, and who need to develop and/or maintain
proper gut flora.
SUMMARY
[0004] This disclosure features human milk permeate compositions,
e.g., compositions that include human oligosaccharides, peptides.,
and other small molecules, and methods of making and using such
compositions. The permeate compositions can contain various levels
of nutritional components and can be used in feeding preterm and
full term infants, as well as children and adults with various
disorders and/or diseases. The compositions are generated from,
inter alia, filtered portions of human milk. The present inventors
have found that surprisingly, permeate (which had been thought to
be a waste product lacking significant nutritional value) contains
high biologically active content, including human oligosaccharides.
It was found that the oligosaccharide content of the permeate and
the human milk products described in application U.S. Ser. No.
11/947,580 (2008/0124430), both natural and concentrated, did not
differ substantially with respect to size and composition as
compared to mother's milk. Because the starting material, from
which permeate is obtained, is pooled human milk, permeate and
other processed human milk products (e.g., those described in
application U.S. Ser. No. 11/947,580) can contain more varieties of
oligosaccharides than individual mother's milk. Therefore, the
permeate can be added to non-human (e.g. bovine) and/or human milk
to increase its nutritional and/or immunologic value. The permeate
can also be used to fight infections, treat inflammatory bowel
disease and help develop and maintain proper gut flora. The
permeate can also be diluted or concentrated and used in such forms
as a nutritional supplement. Similar benefits can be obtained from
permeate obtained by processing pooled human milk and administering
it to infants, e.g., premature infants (e.g., pooled human milk
described in application U.S. Ser. No. 11/947,580).
[0005] The methods featured herein are used to process large
volumes of donor milk, e.g., about 75-2,000 liters/lot of starting
material.
[0006] In a first aspect, the present invention provides a
composition comprising milk and a human milk permeate. In one
embodiment, the milk is human milk. In another embodiment, the milk
is non-human milk. In some embodiments, the non-human milk is
bovine milk. In some embodiments, the permeate of the composition
is obtained by filtering liquid from human milk. In some
embodiments, the composition is obtained by filtering liquid from
human skim milk. In some embodiments, the composition further
comprises vitamins and minerals.
[0007] In a second aspect, the present invention provides a
nutritional composition comprising a concentrated or diluted human
milk permeate. In one embodiment, the permeate is obtained by
filtering liquid from human milk. In other embodiments, the
permeate is obtained by filtering liquid from human skim milk. In
some embodiments, the composition further comprises vitamins and
minerals.
[0008] In a third aspect, the present invention provides a method
of making a concentrated or a diluted human milk permeate,
comprising (a) obtaining human milk, (b) separating the milk into
cream and skim; (c) filtering the skim to obtain a permeate; (d)
retaining the permeate; and (e) diluting, concentrating or drying
the permeate. In some embodiments the diluting step in (e)
comprises adding a non-human milk to the permeate. In some
embodiments, the non-human milk is bovine milk. In other aspects of
the invention, the diluting step in (e) comprises adding water or a
buffer to the permeate. In some embodiments, the concentrating step
in (e) comprises reverse osmosis. In some embodiments, the method
of making a concentrated or diluted human milk permeate further
comprises adding vitamins and minerals to the permeate after step
(d). In some embodiments, the filtering in step (c) comprises
ultrafiltration. In some embodiments, the method of making a
concentrated or diluted human milk permeate further includes (f)
reducing the bioburden. In some embodiments, reducing the bioburden
comprises pasteurization or sterile filtration.
[0009] In a fourth aspect, the present invention provides a method
of administering a permeate composition to a subject comprising
human milk oligosaccharides to a subject comprising (a) obtaining
human milk; (b) separating the milk into cream and skim; (c)
filtering the skim to obtain a permeate; (d) retaining the
permeate; and (e) administering the permeate to the subject. In
some embodiments, the subject is a human preterm or full term
infant. In some embodiments, the composition is administered
topically or orally. In some embodiments, the composition is
administered orally via a feeding tube.
[0010] In a fifth aspect, the present invention provides a method
of establishing beneficial gut flora in a subject comprising
administering a permeate composition comprising oligosaccharides
from human milk to a subject. In some embodiments, the subject is a
human. In some embodiments, the subject is a human preterm or full
term infant. In some embodiments, the human preterm or full term
infant is fed formula after birth. In some embodiments, the
composition is administered prior to, concurrently with, or
following other compositions useful for establishing beneficial gut
flora. In some embodiments, the other compositions useful for
establishing beneficial gut flora are probiotic bacteria or plant
polysaccharides. In some embodiments, the composition is
administered in conjunction with a non-human milk formulation. In
some embodiments, the composition is in a mixture with a non-human
milk formulation. In some embodiments, the composition is a human
milk ultrafiltration permeate. In some embodiments, the permeate is
obtained by filtering human milk. In some embodiments, the permeate
is obtained by filtering human skim milk. In some embodiments,
establishing beneficial gut flora comprises populating the gut with
bifidobacteria or lactobacilli or both.
[0011] In a sixth aspect, the present invention provides a method
of treating a subject who has an infection or is at risk of
developing an infection comprising administering a permeate
composition comprising oligosaccharides from human milk to the
subject. In some embodiments, the symptoms of the infection are
caused by bacteria, bacterial toxins or viruses. In some
embodiments, the subject is a human. In some embodiments, the
subject is a human neonate, infant, child or an adult. In some
embodiments, treating comprises ameliorating at least one symptom
of the infection. In some embodiments, treating comprises promoting
the development of beneficial gut bacteria. In some embodiments,
the beneficial gut bacteria are bifidobacteria or lactobacilli or
both.
[0012] In a seven aspect, the invention provides a method of
treating a subject suffering from an inflammatory bowel disease,
the method comprising administering a permeate composition
comprising oligosaccharides from human milk to a subject. In some
embodiments, the inflammatory bowel disease is one or more of
Crohn's disease, irritable bowel disease, ulcerative colitis (UC),
indeterminate colitis, microscopic colitis, collagenous colitis and
pseudomembrenous colitis. In some embodiments, the subject is a
human. In some embodiments, the subject is a human neonate, infant,
child or an adult. In some embodiments, treating comprises
ameliorating at least one symptom of the inflammatory bowel
disease. In some embodiments, treating comprises promoting
development of beneficial gut bacteria. In some embodiments, the
beneficial gut bacteria are bifidobacteria or lactobacilli or
both.
[0013] In an eighth aspect, the invention provides a method of
making a nutritional supplement comprising obtaining a permeate
composition comprising oligosaccharides from human milk and
supplementing a non-human milk formulation with the composition. In
some embodiments, the composition is a concentrated or diluted
human milk permeate composition. In some embodiments, the permeate
composition is obtained by filtering liquid from human milk. In
some embodiments, the permeate composition is obtained from
filtering liquid from the skim portion of human milk.
[0014] In a ninth aspect, the invention provides a method of making
a concentrated or diluted human milk permeate comprising (a)
obtaining human milk; (b) separating the milk into cream and skim;
(c) filtering the skim to obtain a permeate; (d) retaining the
permeate; and (e) diluting, concentrating, or drying the permeate.
In some embodiments, the diluting step (c) comprises adding a
non-human milk composition to the permeate. In some embodiments,
the non-human milk is bovine milk. In some embodiments, the
diluting step in (c) comprises adding deionized water or a buffer.
In some embodiments the concentrating step in (e) comprises reverse
osmosis. In some embodiments, the method further comprises adding
vitamins and minerals to the permeate after step (d). In some
embodiments, the filtering in step (c) comprises ultrafiltering. In
some embodiments, the method further comprises (f) reducing the
bioburden. In some embodiments, reducing the bioburden comprises
pasteurization or sterile filtration.
[0015] All patents, patent applications, and references cited
herein are incorporated in their entireties by reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 is a chart of an embodiment of a method of making
human milk permeate.
[0017] FIG. 2 is a chart showing a summary of oligosaccharide
composition of samples 42, 48, 49, 53, and 54.
[0018] FIG. 3 is a histogram of common human milk oligosaccharides
found in human milk and in samples 42, 48, 54 (all bulk final
product), 53 (ultrafiltration permeate), and 49 (initial pool of
donor milk).
[0019] FIG. 4 is a graph showing a typical mass spectrometric
profile of free oligosaccharides in human milk. Round dots
represent human milk oligosaccharides.
[0020] FIG. 5 is a graph showing mass spectrum profile obtained for
sample 42 (PROLACT+.TM.H2MF.TM.) (See U.S. Ser. No. 11/947,580)
Round dots represent human milk oligosaccharides.
[0021] FIG. 6 is a graph showing mass spectrum profile obtained for
sample 53 (PROLACT+.TM.H2MF.TM. process byproduct or permeate) (See
U.S. Ser. No. 11/947,580). Round dots represent human milk
oligosaccharides.
[0022] FIG. 7 is a graphical representation of the process of
separating permeate from collected whole human milk.
DETAILED DESCRIPTION
[0023] This disclosure features human milk permeate compositions,
e.g., compositions that include human oligosaccharides, peptides,
and other small molecules and methods of making and using such
compositions. The compositions contain various levels of
nutritional components and can be used in feeding of or
administration to preterm and full term infants, as well as
children and adults with various disorders and/or diseases. The
compositions are generated from, inter alia, filtered portions of
human skim milk. The present inventors have found that,
surprisingly, permeate (which had been thought to be a waste
product lacking significant nutritional value) contains high
biologically active content, including human oligosaccharides.
Because the starting material, from which permeate is obtained, is
pooled human milk, permeate can contain more discrete molecular
forms or types of oligosaccharides than individual mother's
milk.
[0024] The permeate can be added to non-human or human milk to
increase its nutritional value. For example, the permeate can be
added to human milk fortifiers and standardized milk compositions
described in application U.S. Ser. No. 11/947,580, filed on Nov.
29, 2007, the contents of which are incorporated herein by
reference in their entirety. The permeate can also be added to
non-human milk formula, e.g., bovine milk formulations or mixtures
of human and non-human milk formulations. Without being bound by
theory, it is believed by the inventors that permeate compositions
comprising human milk oligosaccharides, peptides, and other small
molecules can be used to promote development of beneficial gut
flora in preterm infants or neonates and to maintain proper gut
flora in children and adults. Permeate can also be useful as a
supplement for infants, children, and adults who have or are at
risk of developing various disorders and diseases, e.g., infections
or immune deficiencies. The permeate can be diluted or concentrated
and/or fortified with vitamins and minerals and used in such forms
as a nutritional supplement.
[0025] The methods featured herein are used to process large
volumes of donor milk, e.g., about 75-2,000 liters/lot of starting
material. The compositions of the present disclosure are generated
from human donor milk, e.g., pooled milk, which undergoes rigorous
genetic screening and processing (e.g., to reduce the
bioburden).
[0026] By "permeate" (also referred to herein as "permeate
composition" or a "milk processing byproduct" or "milk byproducts")
is meant a portion of milk that has been processed by filtration,
e.g., ultrafiltration, of human milk, e.g. skim milk. Typically,
the screen size used in ultrafiltration is 1 Kda-1,000 Kda in size.
The liquid passing through the filtration contains a significant
amount of oligosaccharides and is referred to as permeate.
[0027] By "whole milk" is meant milk from which no fat has been
removed.
[0028] By "skim milk" is meant milk from which at least 75% of fat
has been removed.
[0029] The terms "premature", "preterm" and "low-birth-weight
(LBW)" infants are used interchangeably and refer to infants born
less than 37 weeks gestational age and/or with birth weights less
than 2500 gm.
[0030] The term "full term" infant is used to refer to infants born
after 37 weeks gestational age and/or with birth weights greater
than 2500 gm.
[0031] By "bioburden" is meant microbiological contaminants and
pathogens (generally living) that can be present in milk, e.g.,
viruses, bacteria, mold, fungus and the like.
[0032] Permeate Compositions and Methods of Obtaining Said
Compositions
[0033] Permeate compositions featured herein are obtained from
human milk, e.g., pooled donor milk. Methods of obtaining and
screening human donor milk (including qualifying donors) are
described in applications U.S. Ser. No. 11/947,580, and U.S. Ser.
No. 11/526,127 (2007/0098863), the contents of which are
incorporated herein by reference in their entirety.
[0034] FIG. 1 shows one embodiment of a method of obtaining human
milk permeate. As discussed above, donor milk is carefully analyzed
for both identification purposes and to avoid contamination. The
donor milk is frozen and, when desired, thawed and pooled. It is
then screened (step 1 of FIG. 1), e.g., genetically screened, e.g.,
by polymerase chain reaction (PCR). Genetic screening is done to
identify any contaminants, e.g., viral, e.g., HIV-1, HBV, and/or
HCV. The milk then undergoes filtering, e.g., through about a 200
micron filter (step 2), and heat treatment (step 3). For example,
the composition can be treated at about 63.degree. C. or greater
for about 30 minutes or more. In step 4, the milk is transferred to
a separator, e.g., a centrifuge, to separate the cream from the
skim. The skim can be transferred into a second processing tank
where it remains at about 2 to 8.degree. C. until a filtration step
(step 5). Optionally, the cream separated from the skim in step 4,
can undergo separation again to yield more skim.
[0035] Following separation of cream and skim (step 4), a desired
amount of cream can be added to the skim, and the composition
undergoes further filtration (step 5), e.g., ultrafiltration, e.g.,
with a pore size between 1-1000 Kda. This process concentrates the
nutrients in the skim milk by filtering out the what was previously
thought to be generally water or referred to as permeate. The
present inventors have discovered, however, that the permeate
retains a significant amount of oligosaccharides and can itself be
used, e.g., as a nutritional supplement or in other ways described
herein.
[0036] Skim milk can undergo further processing for a human milk
fortifier or standardized human milk composition, as described,
e.g., in U.S. Ser. No. 11/947,580. For example, the skim can be
blended with cream to obtain bulk final product, pasteurized and
processed for bioburden. The permeate is retained (step 6) and can
be used as a nutritional supplement. FIG. 7 also depicts the
production of a "permeate" and a "second permeate," the latter
produced by additional filtration of the "permeate."
[0037] The permeate can be further processed, e.g., concentrated or
diluted and/or pasteurized. The permeate can be frozen and stored
for future use. The permeate can be concentrated by reverse osmosis
or dried using techniques familiar to those versed in the art. The
resulting liquid products could then be pasteurized, sterile
filtered, or subjected to other bioburden reduction steps. The
product would then be filled into the final product container.
[0038] The permeate can also be supplemented with vitamins and/or
minerals, e.g., calcium, chloride, zinc, copper, iron, manganese,
magnesium, phosphorus, potassium, sodium, selenium, chromium,
molybdenum, iodine, taurine, carnitine, choline, vitamin A, vitamin
B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D,
vitamin E, vitamin K, biotin, folic acid, pantothenic acid, niacin,
and m-inositol. Vitamins and minerals are important, e.g., for
proper nutrition and development of an infant. Trace minerals are
associated with cell division, immune function and growth. Some key
trace minerals include copper, magnesium, and iron (which is
important, e.g., for the synthesis of hemoglobin, myoglobin and
iron-containing enzymes). Zinc is needed, e.g., for growth, for the
activity of numerous enzymes, and for DNA, RNA and protein
synthesis. Copper is necessary for, e.g., the activity of several
enzymes. Manganese is needed, e.g., for the development of bone and
cartilage and is important in the synthesis of polysaccharides and
glycoproteins. Vitamin A is a fat-soluble vitamin essential for,
e.g., growth, cell differentiation, vision and proper functioning
of the immune system. Vitamin D is important, e.g., for absorption
of calcium and to a lesser extent, phosphorus, and for the
development of bone. Vitamin E (tocopherol) prevents peroxidation
of polyunsaturated fatty acids in the cell, thus preventing tissue
damage. Folic acid plays a role in, e.g., amino acid and nucleotide
metabolism. Thus, the permeate compositions can be supplemented
with various minerals and/or vitamins, as desired.
[0039] Applications
[0040] The present disclosure features compositions that include
human milk permeate and methods of using such compositions. The
permeate compositions can be obtained, e.g., as described above by
filtering liquid from skim milk. The compositions contain a
significant number of human oligosaccharides, fucosilated and/or
sialilated as described in Example 1 below. Human milk also
contains peptides and other small molecules that can also be
present in the permeate, e.g., can be obtained by varying the pore
size during ultrafiltration. Because permeate is derived from
pooled human milk, it can contain more forms of oligosaccharides
than found in individual mother's milk. The permeate compositions
can be used alone or in conjunction with other milk compositions,
e.g., human milk compositions described in U.S. Ser. No. 11/947,580
and non-human milk formulas. Permeate or human and non-human milk
formulas supplemented with permeate can be administered to pre-term
or full term infants to, e.g., promote development of proper gut
flora, treat conditions characterized by immunodeficiency, and
treat or prevent infections. Permeate can also be administered to
children and adults alone or in conjunction with probiotics to
promote beneficial gut flora, treat conditions characterized by
immunodeficiency, and treat or prevent infections. Permeate
compositions can be administered, e.g., orally or topically to
treat skin conditions.
[0041] Human Milk Oligosaccharides
[0042] Many factors can affect the clinical outcome of a newborn,
e.g., prematurely born, infant. Preterm infants have vulnerable
immune systems, immature digestive systems, and increased total
caloric and specific nutrient needs (when generally compared with
full term infants). Thus, nutrition provided to such infants is an
important factor in their growth and development. Human milk has
long been recognized as the ideal food for preterm and full term
infants because of its nutritional composition and immunologic
benefits. Not every mother, however, can or will breastfeed her
baby (or use a breast pump and store her milk). For example,
mothers who have certain diseases, e.g., active tuberculosis, or
are being administered radioisotopes, antimetabolites, or
chemotherapy may not breast feed their infants. In addition,
mother's own milk may not contain sufficient nutritional content to
sustain a preterm infant. Use of donor milk can also be
problematic, as such milk may not contain adequate nutrition for a
preterm infant.
[0043] The present disclosure features permeate compositions that
include oligosaccharides from pooled human milk, and methods of
using such compositions to benefit, e.g., of premature human
infants, full term infants, children, and adults.
[0044] Human milk oligosaccharides are carbohydrates that contain
lactose at the reducing end and, typically, a fucose or a sialic
acid at the nonreducing end. (Morrow et al., J. Nutrit.
135:1304-07, 2005). These terminal sugars are the residues that
most strongly influence the selective growth of bacteria and the
interaction of oligosaccharides with other molecules or cells,
including bacterial pathogens in the gut lumen. For example, sialic
acids are structural and functional components of brain
gangliosides and have been implicated in neurological development
of infants. Oligosaccharides can be free or conjugated as
glycoproteins, glycolipids etc. and are classified as glycans. They
constitute the third most numerous solid component of human milk,
after lactose and lipid (Morrow et al.). The majority of these milk
oligosaccharides, however, are not digcstablc by infants and can be
found in infant feces largely intact. The present disclosure
describes various uses of these oligosaccharides present in human
milk permeate (which, in fact, contains a wide variety of
oligosaccharides as it is derived from a pool of donated milk),
which was previously thought to be a waste product of processing
human milk. The permeate or compositions obtained from the permeate
can be used (alone or in combination with human and non-human milk
formulations or in combination with drugs or prebiotics), e.g., to
promote beneficial gut flora, fight infections, ameliorate symptoms
of conditions characterized by immunodeficiency, and provide
additional nutrition. These compositions can be beneficial to
recipients of various ages: from preterm infants through children
to adults.
[0045] Gut Flora
[0046] Human gut flora, i.e., bacteria found in the gut, serves
various functions, including digestion of certain polysaccharides
and development of the gut's mucosal immune system. Gut bacteria
can stimulate the lymphoid tissue associated with the gut mucosa to
produce antibodies to pathogens, leaving the helpful species alone,
a tolerance developed in infancy.
[0047] The gastrointestinal tract of a normal fetus is sterile.
During birth and soon thereafter, the infant's gut is colonized by
various bacteria. The sources of the bacteria can be the mother's
body and/or the environment. After birth, various bacteria can be
transferred from the mother to the infant through suckling and
touching. Most infant GI tracts arc initially colonized by large
numbers of E. coli and Streptococci. Eventually, breast-fed babies
become dominated by bifidobacteria, while formula-fed infants have
a variety of bacteria, including Enterobacteriaceae, Enterococci,
bifidobacteria, Bacteroides, and Clostridia in the gut. After the
introduction of solid food and weaning, the microflora of
breast-fed infants becomes similar to that of formula-fed infants.
By about two years, the fecal microflora of children is similar to
that of adults.
[0048] Given the importance of gut flora, it is crucial to develop
a beneficial bacterial population at birth and maintain it
throughout childhood and adult life. The compositions and methods
featured herein can help in the establishment and maintenance of
such beneficial gut flora.
[0049] The compositions that include human milk oligosaccharides,
e.g., human milk permeate or compositions derived from human milk
permeate, can be administered to preterm infants, full term
infants, children, and adults. They can be administered alone or in
combination with other compositions that aid in establishing
beneficial gut flora, e.g., probiotic bacteria or prebiotic plant
polysaccharides, to form a symbiotic composition. They can be
administered as part of human milk formulations (e.g., compositions
described in application U.S. Ser. No. 11/947,580) or non-human
milk baby formulas. Without being bound by theory, it is believed
that the present permeate compositions can promote colonization of
the gut by lactobacilli and/or bifidobacteria. Both lactobacilli
and bifidobacteria are known as probiotics--bacteria that protect
their host and prevent disease. Bifidobacteria are anaerobic
bacteria that aid in digestion and have been associated with
reduced incidence of allergies and cancer prevention. Lactobacilli
are bacteria that convert lactose and other sugars to lactic acid
and may possess anti-inflammatory and anti-cancer properties.
[0050] Additionally, the human milk permeate or compositions
derived from human milk permeate of the current invention
containing human milk oligosaccharides can be administered to
children or adults for the treatment of inflammatory bowel disease.
Inflammatory bowel disease is a collective term encompassing
related, but distinct, chronic inflammatory disorders of the
gastrointestinal tract, such as Crohn's disease, irritable bowel
disease, ulcerative colitis, indeterminate colitis, microscopic
colitis, and collagenous colitis. It has been found that patients
suffering from active Crohn's disease have significantly less
recoverable bifidobacteria in their feces compared with healthy
individuals. This reduction in bifidobacteria numbers was observed
to be directly correlated with decreased levels off3-D
galactosidase production and activity (Favier, C. et at, Dig. Dis.
Sci. 1997;42:817-822). 13-D galactosidase is an enzyme produced by
bifidobacteria. These results support suggestions proposed in other
studies that strains of bifidobacteria may play important roles in
maintaining a balanced healthy intestinal microflora. Therefore,
according to the present invention, the colony promotion of
colonization of bifidobacteria in the gut by the oligosaccharide
rich permeate of the current invention can be useful in
ameliorating the symptoms of inflammatory bowel disease. Similarly,
the compositions of the current invention can be useful in treating
pseudomembrenous colitis (also known as C. difficile-related
colits), a common complication of prolonged broad-spectrum
antibiotic treatment that is due to overgrowth of normal flora by a
toxin-producing strain of C. difficile. The permeate compositions
of the current invention can be administered before during or after
any other treatment for inflammatory bowel disease including
probiotic formulations, anti-inflammatory medications or
immunomodulators.
[0051] Infections
[0052] Human milk oligosaccharides are thought to serve various
anti-infective functions, e.g., inhibit pathogen binding. Thus, the
permeate compositions featured herein and containing a wide variety
of human milk oligosaccharides (as they are derived from pooled
human milk) can be used to protect infants, children, and adults
from various types of infection. For example, preterm and full term
infants administered the present compositions can be protected from
infectious diarrhea, necrotizing enterocolitis (NEC), respiratory
tract infections, bacteremia, meningitis and others. Children and
adults can likewise benefit from administration of the
compositions. The compositions can be administered after an onset
of an infection, to ameliorate and/or eliminate its symptoms or
prior to infection (for individuals thought to be at risk). The
compositions can be administered via various means, including
orally or topically (e.g., to treat skin infections and other skin
conditions).
[0053] In general, the permeate compositions featured herein can be
administered to subjects, e.g., human subjects of various ages, for
any condition or disorder that would be ameliorated or eliminated
by application of oligosaccharides, peptides, or other small
molecules derived from human milk.
[0054] Kits
[0055] The present disclosure also features kits that include the
human milk permeate compositions described herein and containers
for mixing the compositions with raw human milk or non-human milk.
The containers can include bottles, e.g., graduated bottles to
assist in proper dilution, syringes, cans, and other containers
known in the art.
[0056] The embodiments of the disclosure may be carried out in
other ways than those set forth herein without departing from the
spirit and scope of the disclosure. The embodiments are, therefore,
to be considered in all respects as illustrative and not
restrictive.
EXAMPLE 1
Collection and Analysis of Human Milk Permeate
[0057] Breast milk was collected from screened donors throughout
the U.S. and stored at -20.degree. C. for up to 12 months. Pooled
donor human milk was processed and various samples analyzed for
their oligosaccharide composition. Pools from 20-50 donors were
used to produce human milk fortifier compositions described in
application U.S. Ser. No. 11/947,580.
[0058] Before processing, a sample of initial pooled donor milk was
set aside for analysis (sample 49 in Table III). The pooled milk
was screened, filtered, heat-treated, separated into skim and
cream, and the skim was ultrafiltered. A portion of the
filtered-out composition, the permeate, was set aside for analysis
(sample 53 in Table IV). The skim was blended with cream and
pasteurized. A portion of the bulk final product was set aside for
analysis (samples 42, 48, and 54 in Tables I, II, and V,
respectively).
[0059] Each milk sample that was set aside for analysis (0.5 mL)
was diluted with 0.5 mL of pure water and centrifuged at 4000 rpm
at 4.degree. C. for 30 minutes to separate fat. The permeate was
analyzed undiluted. The fat-free fraction was treated with 4
volumes (2:1) of a chloroform-methanol solution (v/v). The emulsion
was centrifuged at 3500 rpm for 30 min at 4.degree. C., and the
lower chloroform layer and denatured protein were discarded. The
upper layer was collected, two volumes of pure ethanol were added
and protein fraction was left to precipitate at 4.degree. C.
overnight.
[0060] After protein separation by centrifugation at 3500 rpm for
30 min at 4.degree. C., the limpid upper solution was collected and
freeze-dried. The resulting powder (freeze-dried oligosaccharide
rich fraction) was used for oligosaccharide analysis.
Oligosaccharides were reduced to alditol form using 1.0 M sodium
borohydride in deionized water and incubated at 42.degree. C.`
overnight. After the reaction, oligosaccharides were purified from
contaminants by solid-phase extraction using a nonporous
graphitized carbon cartridge (GCC-SPE). Nonporous graphitized
carbon cartridges (150 mg of bed weight, 4 mL tube size) for
desalting were purchased from Alltech (Deerfield, Ill., USA).
Evaporation of solvents was performed using a speedvac
centrifuge.
[0061] Solid Phase Extraction. Prior to use, each GCC-SPE cartridge
was washed with 3 column volumes of 80% acetonitrile in 0.05%
trifluoroacetic acid (TEA) (v/v) followed by 3 column volumes of
deionized water. After loading of the oligosaccharide mixture onto
a cartridge, salts and residual peptides were removed by washing
with 8 cartridge volumes of deionized water. The oligosaccharides
were then eluted from the column using 10% acetonitrile in water
(v/v) and 20% acetonitrile in water (v/v). Each fraction (6 mL) was
collected and evaporated in vacuo prior to MS analysis.
[0062] Mass Spectrometric Analysis. Matrix-assisted laser
desorption/ionization (MALDI) Fourier transform-ion cyclotron
resonance mass spectrometry (FT ICR MS) was performed on an
HiResMALDI (lonSpec Corp., Irvine, Calif.) equipped with an
external MALDI source, a 7.0 Tesla superconducting magnet and a
pulsed Nd:YAG laser (355 nm). 2,5-Dihydroxybenzoic acid (DHB) was
used as a matrix (5 mg/100 .mu.L in a solution of 50%
acetonitrile/50% water (v/v)). The solution of oligosaccharide (1
.mu.L) was applied to the MALDI probe followed by addition of 0.01
M NaCl (0.5 .mu.L) and the matrix solution (1 .mu.L). The sample
was dried under a stream of air and subjected to mass
spectrometry.
[0063] The compositions of the various samples discussed above are
presented in Tables I-IV, and a summary is shown in FIG. 2. The
experimental mass:charge ratios (m/z expe) were matched to
calculated mass:charge ratios (m/z cal) for known oligosaccharides
in human milk. Mass spectrometric profiles of specific samples arc
shown in Tables III-V (human milk sample in Table III, bulk product
in Table IV, and permeate in Table V). Mass spectrometric analyses
of the five samples revealed a highly complex mixture of
oligosaccharides, typical of human milk, and varying in size,
composition, and abundance.
[0064] The oligosaccharides identified were from two main classes:
(1) neutral oligosaccharides containing galactose,
N-acetylglucosamine, lactose, and fuctose (Hex, HexNAc and Fuc);
and (2) anionic oligosaccharides containing the same
oligosaccharide compositions with the addition of
N-acetylneuraminic acid (NeuAC). A previous investigation
(Ninonuevo et al., J. Agric. Food Chem. 2008, 54: 7471-7480)
identified diversity between individuals in the total numbers and
relative abundances of specific oligosaccharides in unfiltered
milk. Thus, the milk samples analyzed (natural or concentrated)
confirmed that all samples show ostensibly most of the
oligosaccharides previously found to be common in human milk from
different donors.
TABLE-US-00001 TABLE I SAMPLE 42 (BULK FINAL PRODUCT)
OLIGOSACCHARIDES m/z expe Hex HexNAc Fuc NeuAC m/z cal Error Mode
Form abund. 1389.477 4 2 2 0 1389.501 -0.0246 Na Alditol 100
1243.419 4 1 2 0 1243.443 -0.0239 Na Alditol 79.32 1754.611 5 2 3 0
1754.633 -0.0228 Na Alditol 36.36 1608.563 5 1 3 0 1608.576 -0.013
Na Alditol 22.4 1900.682 5 3 3 0 1900.691 -0.0095 Na Alditol 18.22
1535.545 4 3 2 0 1535.559 -0.0144 Na Alditol 17.4 878.3013 3 1 1 0
878.3111 -0.0098 Na Alditol 15.74 1097.368 4 0 2 0 1097.385 -0.0176
Na Alditol 14.48 732.245 3 0 1 0 732.2532 -0.0082 Na Alditol 13.02
1024.361 3 2 1 0 1024.369 -0.0083 Na Alditol 6.88 1462.514 5 0 3 0
1462.518 -0.0041 Na Alditol 4.29 1556.519 4 1 2 1 1556.521 -0.0021
1 Na Alditol 3.48 2119.781 6 2 4 0 2119.766 0.0156 Na Alditol 2.72
2046.765 5 4 3 0 2046.749 0.0154 Na Alditol 2.24 1557.524 4 3 2 0
1557.541 -0.0168 1 Na Alditol 2.21
TABLE-US-00002 TABLE II SAMPLE 48 (BULK FINAL PRODUCT)
OLIGOSACCHARIDES m/z expe Hex HexNAc Fuc NeuAC m/z cal Error Mode
Form abund. 1389.474 4 2 2 0 1389.501 -0.0274 Na Alditol 100
1754.609 5 2 3 0 1754.633 -0.0243 Na Alditol 15.63 1462.519 3 5 1 0
1462.543 -0.0239 Na Alditol 1.27 1243.422 4 1 2 0 1243.443 -0.0212
Na Alditol 73.84 1405.476 5 1 2 0 1405.496 -0.0202 Na Alditol 1.45
1900.672 5 3 3 0 1900.691 -0.0198 Na Alditol 10.32 1535.54 4 3 2 0
1535.559 -0.0196 Na Alditol 23.25 1097.373 4 0 2 0 1097.385 -0.0129
Na Alditol 11.85 1608.567 5 1 3 0 1608.576 -0.0089 Na Alditol 6.79
1024.364 3 2 1 0 1024.369 -0.0047 Na Alditol 14.85 878.3075 3 1 1 0
878.3111 -0.0036 Na Alditol 24.05 2046.746 5 4 3 0 2046.749 -0.0031
Na Alditol 1.28 732.2536 3 0 1 0 732.2532 0.0004 Na Alditol 14.72
1462.519 5 0 3 0 1462.518 0.0012 Na Alditol 1.27 1081.392 3 1 2 0
1081.391 0.0015 Na Alditol 1.99 513.1818 2 1 0 0 513.1789 0.0029 Na
Alditol 9.19 935.3364 3 0 2 0 935.3326 0.0038 Na Alditol 1.26
659.2407 2 2 0 0 659.2368 0.0039 Na Alditol 1.08
TABLE-US-00003 TABLE III SAMPLE 49 (INITIAL POOL OF DONOR MILK)
OLIGOSACCHARIDES m/z expe Hex HexNAc Fuc NeuAC m/z cal Error Mode
Form abund. 1388.972 4 3 3 0 1388.99 -0.0202 Na Alditol 100
1754.608 5 2 3 0 1754.633 -0.0258 Na Alditol 5.1 1243.423 4 1 2 0
1243.443 -0.0207 Na Alditol 4.1 1900.671 5 3 3 0 1900.691 -0.0202
Na Alditol 1.48 1535.542 4 3 2 0 1535.559 -0.0169 Na Alditol 5.9
1608.562 5 1 3 0 1608.576 -0.0139 Na Alditol 3.6 1097.377 4 0 2 0
1097.385 -0.0084 Na Alditol 10.5 1024.364 3 2 1 0 1024.369 -0.0054
Na Alditol 14.7 1462.513 5 0 3 0 1462.518 -0.0042 Na Alditol 1.2
878.3073 3 1 1 0 878.3111 -0.0038 Na Alditol 26 732.2535 3 0 1 0
732.2532 0.0003 Na Alditol 14.2 513.1826 2 1 0 0 513.1789 0.0037 Na
Alditol 8.9 659.241 2 2 0 0 659.2368 0.0042 Na Alditol 1.66
TABLE-US-00004 TABLE IV SAMPLE 53 (PERMEATE) OLIGOSACCHARIDES m/z
expe Hex HexNAc Fuc NeuAC m/z cal Error Mode Form abund. 1973.685 6
1 4 0 1973.708 -0.0229 Na Alditol 100 2119.75 6 2 4 0 2119.766
-0.0152 Na Alditol 89.8 1608.564 5 1 3 0 1608.576 -0.0116 Na
Alditol 69.33 1754.626 5 2 3 0 1754.633 -0.0077 Na Alditol 54.88
1827.651 6 0 4 0 1827.65 0.0009 Na Alditol 41.26 2265.835 6 3 4 0
2265.824 0.0119 Na Alditol 34.5 1462.52 5 0 3 0 1462.518 0.0022 Na
Alditol 28.01 1900.709 5 3 3 0 1900.691 0.0177 Na Alditol 15.16
2484.931 7 2 5 0 2484.898 0.0334 Na Alditol 15.12 2338.868 7 1 5 0
2338.84 0.028 Na Alditol 14.2 1243.447 4 1 2 0 1243.443 0.0039 Na
Alditol 13.39 1389.508 4 2 2 0 1389.501 0.0069 Na Alditol 11.41
732.2539 3 0 1 0 732.2532 0.0007 Na Alditol 8.37 2631.035 7 3 5 0
2630.956 0.0792 Na Alditol 4.85 2192.827 7 0 5 0 2192.782 0.0447 Na
Alditol 4.82 2411.932 6 4 4 0 2411.881 0.0504 Na Alditol 3.39
2704.042 8 1 6 0 2703.972 0.0697 Na Alditol 2.85 1556.543 4 1 2 1
1556.521 0.0227 Na Alditol 2.67 2850.121 8 2 6 0 2850.03 0.0908 Na
Alditol 2.16 570.2012 2 0 1 0 570.2004 0.0008 Na Alditol 1.94
1097.392 4 0 2 0 1097.385 0.0063 Na Alditol 1.82 2557.988 8 0 6 0
2557.914 0.0739 Na Alditol 1.28 1300.476 4 0 3 0 1300.465 0.011 Na
Alditol 1.19 659.2383 2 2 0 0 659.2368 0.0015 Na Alditol 1.16
513.1802 2 1 0 0 513.1789 0.0013 Na Alditol 1.01
TABLE-US-00005 TABLE V SAMPLE 54 (BULK FINAL PRODUCT)
OLIGOSACCHARIDES m/z expe Hex HexNAc Fuc NeuAC m/z cal Error Mode
Form abund. 513.1576 2 1 0 0 513.1789 -0.0213 Na Alditol 12.82
659.2137 2 2 0 0 659.2368 -0.0231 Na Alditol 1.38 570.1783 2 0 1 0
570.2004 -0.0221 Na Alditol 4.24 878.2782 3 1 1 0 878.3111 -0.0329
Na Alditol 20.97 1024.338 3 2 1 0 1024.369 -0.0309 Na Alditol 8.82
732.2245 3 0 1 0 732.2532 -0.0287 Na Alditol 16 935.3093 3 0 2 0
935.3326 -0.0233 Na Alditol 1.16 1243.39 4 1 2 0 1243.443 -0.0536
Na Alditol 76.72 1389.445 4 2 2 0 1389.501 -0.0567 Na Alditol 100
1535.524 4 3 2 0 1535.559 -0.0348 Na Alditol 19.2 1097.334 4 0 2 0
1097.385 -0.0511 Na Alditol 14.86 1608.547 5 1 3 0 1608.576 -0.029
Na Alditol 19.86 1754.592 5 2 3 0 1754.633 -0.0416 Na Alditol 36.7
1900.669 5 3 3 0 1900.691 -0.0225 Na Alditol 24.32 2046.773 5 4 3 0
2046.749 0.0236 Na Alditol 4.32 1462.497 5 0 3 0 1462.518 -0.0204
Na Alditol 3.8 1973.729 6 1 4 0 1973.708 0.0213 Na Alditol 1.66
2119.792 6 2 4 0 2119.766 0.0261 Na Alditol 4.15 2265.855 6 3 4 0
2265.824 0.0314 Na Alditol 4.45 2411.941 6 4 4 0 2411.881 0.0596 Na
Alditol 1.3 m/z expe: Mass/charge experimental, m/z cal:
mass/charge calculated, Hex: hexose; HexNAc: N-acetylhexosamine;
Fuc: fucose; NeuAc: N-acetyl-neuraminic acid or sialic acid;
[0065] It appears that processed human milk product (fortifier or
samples 42, 48, and 54) and permeate (sample 53) contain the full
range of oligosaccharides commonly found in human milk. The
selection and amounts of the individual oligosaccharides present in
these samples reflect the relative amounts in the donor pool which
average out the variation across individual donors.
[0066] Thus, according to the present invention, the permeate is a
valuable source of human milk oligosaccharides that can be used as
a nutritional or immunologic supplement for preterm infants,
infants, children, and adults.
EXAMPLE 2
Oligosaccharide Profile of Pooled Permeate Samples
[0067] Six pooled permeate samples were analyzed for specific
oligosaccharide content (Table VI). Each of the pooled permeate
samples were derived from pooled milk received from multiple donors
by the method discussed above in Example 1. The collected samples
of permeate were analyzed by mass spectrometry by the method
described above in Example 1, to detect the presence (1) or absence
(0) of the specific oligosaccharides indicated. The
oligosaccharides arc represented in Table VI by their mass/charge
ratios.
TABLE-US-00006 TABLE VI Per- Per- Per- Olgios meate Permeate meate
Permeate meate Permeate m/z Sample Sample Sample Sample Sample
Sample expe #5 #11 #23 #28 #47 #53 513 0 0 1 0 1 0 732 1 1 1 1 1 1
878 1 1 1 1 1 1 935 0 0 1 0 0 1 1024 1 1 1 1 1 0 1097 1 1 1 1 1 1
1243 1 1 1 1 1 1 1389 1 1 1 1 1 1 1462 1 1 1 0 1 1 1535 0 0 1 0 1 0
1556 0 0 0 0 0 1 1608 1 0 1 1 1 1 1754 0 1 1 1 0 1 1827 1 0 0 0 0 1
1900 0 1 0 0 0 1 1973 1 0 0 1 0 1 2119 1 1 0 1 0 1 2192 0 0 0 0 0 1
2264 0 0 0 0 0 1 2338 0 0 0 0 0 1 2411 0 0 0 0 0 1 2485 0 0 0 0 0 1
2631 0 0 0 0 1 1 = oligosaccharide present in the sample 0 =
oligosaccharide not detectable in the sample
[0068] It appears as though, while there is overlap between the
samples with respect to certain oligosaccharides (i.e.
oligosaccharides 732, 878, 1097, 1243 and 1389 are present in all
six samples), there are also many oligosaccharides present in
certain samples while not present in other samples (i.e.
oligosaccharide 935 is present in permeate numbers 23, and 53 but
not permeate numbers 5, 11, 28 or 47). Therefore, additional
pooling of the pooled permeate samples may be necessary in order to
obtain a composition with substantially all or all oligosaccharide
species represented.
EXAMPLE 3
Administration of Permeate Compositions
[0069] As discussed above, it is believed by the inventors that
human milk permeate compositions from pooled milk can provide a
wide variety of nutritional and immunologic benefits, as they are a
source of a wider variety of oligosaccharides than those found in
individual mother's milk. They can also contain various types of
peptides and other beneficial small molecules.
[0070] The isolated human milk permeate compositions will be
administered to infants, both preterm and full term infants, to
promote their development of proper gut flora, ameliorate and/or
eliminate infections. The study will investigate infants divided
into 5 arms including infants receiving:
[0071] 1. Mother's milk plus fortifier
[0072] 2. Cow's milk plus permeate
[0073] 3. Cow's milk plus strain 1 of bifidobacter
[0074] 4. Cow's milk plus strain 2 of bifidobacter
[0075] 5. Cow's milk plus non-milk derived fructose
oligosaccharide
[0076] Composition of infant feces will be monitored to determine
the content of their gut flora. Infants suffering from infections
caused by pathogens, e.g., bacteria, bacterial toxins, and viruses
will particularly benefit from oral administration of the permeate
compositions. It is expected that the gut flora from infants
receiving cow's milk plus permeate will most closely mimic the gut
flora from infants receiving mother's milk plus fortifier.
[0077] Other variations and embodiments of the invention described
herein will now be apparent to those of ordinary skill in the art
without departing from the scope of the invention or the spirit of
the claims below.
* * * * *