U.S. patent application number 16/681626 was filed with the patent office on 2020-05-14 for composition and method for treating acute respiratory tract infections.
The applicant listed for this patent is Glycom A/S. Invention is credited to Bruce McConnell, Louise Kristine Vigsn.ae butted.s.
Application Number | 20200147114 16/681626 |
Document ID | / |
Family ID | 56849164 |
Filed Date | 2020-05-14 |
United States Patent
Application |
20200147114 |
Kind Code |
A1 |
McConnell; Bruce ; et
al. |
May 14, 2020 |
COMPOSITION AND METHOD FOR TREATING ACUTE RESPIRATORY TRACT
INFECTIONS
Abstract
A composition consists of an effective amount of a synthetic
neutral fucosylated human milk oligosaccharide ("HMO"), an
effective amount of a synthetic neutral non-fucosylated HMO, and
one or more excipients acceptable for enteral administration to a
non-infant human. In some embodiments, the synthetic neutral
fucosylated HMO is 2'-fucosyllactose and the synthetic neutral
non-fucosylated HMO is selected from lacto-N-neotetraose ("LNnT")
and lacto-N-tetraose ("LNT"). In certain embodiments, the mass
ratio of 2'-FL to the selected LNnT or LNT is about 1.5 to about
4.5. In one embodiment, the synthetic neutral non-fucosylated HMO
is lacto-N-neotetraose ("LNnT"). In certain embodiments, the one or
more excipients acceptable for enteral administration to a human
are selected from diluents, antioxidants, lubricants, colorants,
binders, disintegrants, and combinations thereof. In various
embodiments, the one or more excipients are effective to form a
galenic unit dosage form selected from a capsule, a tablet, and a
sachet.
Inventors: |
McConnell; Bruce; (La Tour
de Peilz, CH) ; Vigsn.ae butted.s; Louise Kristine;
(Kobenhavn, DK) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Glycom A/S |
Horsholm |
|
DK |
|
|
Family ID: |
56849164 |
Appl. No.: |
16/681626 |
Filed: |
November 12, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15555936 |
Sep 5, 2017 |
10471084 |
|
|
PCT/DK2016/050062 |
Mar 4, 2016 |
|
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16681626 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/702 20130101;
A23V 2002/00 20130101; A23L 33/125 20160801; A61K 31/7048 20130101;
A61K 9/205 20130101; A61P 31/00 20180101; A61P 11/04 20180101; A61K
9/2054 20130101; A23V 2002/00 20130101; A23V 2200/3202 20130101;
A61K 31/702 20130101; A23V 2250/28 20130101; A23V 2200/314
20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/7048 20060101
A61K031/7048; A23L 33/125 20060101 A23L033/125; A61K 9/20 20060101
A61K009/20; A61K 31/702 20060101 A61K031/702 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 5, 2015 |
DK |
PA201570124 |
Feb 24, 2016 |
DK |
PA201670097 |
Claims
1. A composition consisting of: an effective amount of a synthetic
neutral fucosylated human milk oligosaccharide ("HMO") consisting
of 2'-fucosyllactose ("2'-FL"); an effective amount of at least one
synthetic neutral non-fucosylated HMO selected from
lacto-N-neotetraose ("LNnT") and lacto-N-tetraose ("LNT"); and one
or more excipients acceptable for enteral administration to a
human.
2. (canceled)
3. The composition of claim 1, wherein the mass ratio in the
composition of 2'-FL to the at least one synthetic neutral
non-fucosylated HMO is about 1.5 to about 4.5.
4. The composition of claim 1, wherein the synthetic neutral
non-fucosylated HMO consists of lacto-N-neotetraose ("LNnT").
5. The composition of claim 1, wherein the one or more excipients
acceptable for enteral administration to the human are selected
from diluents, antioxidants, lubricants, colorants, binders,
disintegrants, and combinations thereof.
6. The composition of claim 1, wherein the one or more excipients
are effective to form a galenic unit dosage form selected from a
capsule, a tablet, and a sachet.
7. A composition consisting essentially of: an effective amount of
synthetic 2'-fucosyllactose ("2'-FL") and at least one synthetic
neutral human milk oligosaccharide ("HMO") selected from
lacto-N-neotetraose ("LNnT") and lacto-N-tetraose ("LNT").
8. The composition of claim 7, wherein the mass ratio in the
composition of 2'-FL to the at least one synthetic neutral
non-fucosylated HMO is about 1.5 to about 4.5.
9. The composition of claim 7, wherein the at least one synthetic
neutral non-fucosylated HMO is lacto-N-neotetraose ("LNnT").
10. A composition consisting of: an effective amount of synthetic
2'-fucosyllactose ("2'-FL") and at least one synthetic neutral
human milk oligosaccharide ("HMO") selected from
lacto-N-neotetraose ("LNnT") and lacto-N-tetraose ("LNT").
11. The composition of claim 10, wherein the mass ratio in the
composition of 2'-FL to the at least one synthetic neutral
non-fucosylated HMO is about 1.5 to about 4.5.
12. The composition of claim 11, wherein the at least one synthetic
neutral non-fucosylated HMO is lacto-N-neotetraose ("LNnT").
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This is a continuation application of and claims priority to
U.S. patent application Ser. No. 15/555,936 entitled "COMPOSITION
AND METHOD FOR TREATING ACUTE RESPIRATORY TRACT INFECTIONS" and
filed on Sep. 5, 2017 for Bruce McConnell, et al., and claims the
priority to PCT/DK2016/050062 entitled "COMPOSITION AND METHOD FOR
TREATING ACUTE RESPIRATORY TRACT INFECTIONS" and filed on Mar. 4,
2016 for Bruce McConnell, et al., and claims priority to Denmark
Patent Application No. PA201570124 filed on Mar. 5, 2015 and claims
priority to Denmark Patent Application No. PA201670097 filed on
Feb. 24, 2016, which are incorporated herein by reference for all
purposes.
FIELD OF THE INVENTION
[0002] This invention relates to a composition for use in
preventing an acute respiratory tract infection (ARI) and/or
relieving symptoms of an ARI in immune-compromised persons,
particularly adults. The invention also relates to a method for the
treatment or prevention of an ARI in immune-compromised persons
BACKGROUND
[0003] Acute infections of the respiratory tract are very common,
especially in infants, children and the elderly. In the US, for
patients five years or older, ARI's accounted for 8% of all visits
to primary care and 58% of all antibiotics prescribed in primary
care (Grijalva et al. JAMA 302, 758 (2009)). These infections
produce conditions such as otitis, bronchitis, pneumonia,
sinusitis, pharyngitis and strep throat.
[0004] Acute bronchitis, which results from an infection, is an
inflammation of the bronchi. It is one of the most prevalent
respiratory infections treated in primary care and occurs most
often during the winter. It is an acute illness usually lasting
less than three weeks with coughing as the main symptom and at
least one other lower respiratory tract symptom such as wheezing,
sputum production or chest pain. Older people can have symptoms
such as confusion or rapid breathing. About 90% of cases of acute
bronchitis are caused by viruses, such as rhinoviruses,
coronaviruses, adenoviruses, metapneumoviruses, parainfluenza
viruses, respiratory syncytial viruses and influenza viruses. About
10% of cases are caused by bacteria such as Mycoplasma pneumoniae,
Chlamydophila pneumoniae, Bordetella pertussis, Streptococcus
pneumonia and Haemophilus influenzae.
[0005] Treatment for acute bronchitis usually involves just
treating the symptoms, e.g., with non-steroidal anti-inflammatory
drugs (NSAIDs) to treat fever and sore throat. As most cases of
acute bronchitis are caused by viruses, antibiotics are not
recommended but tend to be heavily overused. Over 60% of bronchitis
patients are prescribed antibiotics, and this is a leading cause of
the development of antibiotic-resistant bacteria. This over
prescription occurs despite fact that there is no clinical benefit
for most patients.
[0006] While acute bronchitis is generally not serious, serious
complications can occur as a result. Pneumonia is the most common
complication of acute bronchitis and occurs when the infection
spreads further into the lungs, causing air sacs inside the lungs
to fill up with fluid. 1 in 20 cases of acute bronchitis leads to
pneumonia. Another possible complication is respiratory failure. It
is also possible that acute bronchitis can lead to chronic
bronchitis. These complications are more common in
immune-compromised people such as the elderly, diabetics, etc. For
example, the number of hospitalisations for acute lower respiratory
infections in England is about three times higher in those over 75
years than in younger people. Further, the average length of stay
for acute respiratory conditions increases progressively with
age.
[0007] Although most ARI's are resolved reasonably quickly, the
suffering and the financial costs of respiratory tract infections
are enormous. Little effective prevention is currently possible
outside of two strategies: attempting to avoid contact with or
spreading of infectious agents; and vaccination for influenza and
pneumococcal pneumonia.
[0008] Human milk oligosaccharides (HMOs) have been proposed as a
possible means for preventing or treating ARIs. In WO 2012/076323,
a method is disclosed for preventing or treating ARIs in children
of up to three years old using HMOs. The children are treated using
nutritional compositions which contain an N-acetyl lactosamine, a
sialylated oligosaccharide and a fucosylated oligosaccharide. The
method is in particularly suited for treating otitis or
bronchiolitis. In WO 2014/100696, a nutritional composition is
disclosed for modulating inflammation, such as a respiratory
virus-induced inflammation, in adults and the elderly. In US
2015/0031645, a method is disclosed for improving the immune
system's response to a viral infection, such as a respiratory viral
infection, in children of up to twelve years old. A nutritional
composition which contains 2'-fucosyllactose and
lacto-N-neotetraose is disclosed as being preferred.
[0009] However, there remains a need for an effective, as well as
convenient and safe, method for preventing ARI infections and/or
relieving their symptoms in immune-compromised persons,
particularly adults.
SUMMARY
[0010] A composition in one embodiment may be used for in
preventing and/or relieving symptoms of an acute respiratory tract
infection (ARI) in immune-compromised persons, particularly adults,
more particularly pregnant women and the elderly, the composition
comprising 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose (LNnT)
and/or lacto-N-tetraose (LNT).
[0011] In some embodiments, the composition consists essentially of
2'-FL and LNnT and/or LNT. In various embodiments, the composition
contains more 2'-FL than LNnT and/or LNT. Even more advantageously,
the mass ratio of 2'-FL to LNnT and/or LNT in the composition is
about 1.5 to about 4.5. In one embodiment, the mass ratio of 2'-FL
to LNnT and/or LNT in the composition is about 1.5 to about 2.5,
for example about 2:1. In another embodiment, the mass ratio of
2'-FL to LNnT and/or LNT in the composition is about 3.5 to about
4.5, for example about 4:1.
[0012] In one embodiment, the composition comprises at least 0.5 g
of 2'-FL and LNnT and/or LNT, in other embodiments the composition
comprises at least 1 g of 2'-FL and LNnT and/or LNT, for example
about 1.5 g or more of 2'-FL and LNnT and/or LNT.
[0013] In certain embodiments, the composition is a once-a-day
formulation providing at least about 2 g of 2'-FL and LNnT and/or
LNT per day, more advantageously at least 3 g of 2'-FL and LNnT
and/or LNT.
[0014] A method of reducing a risk of acute respiratory tract
infection and/or relieving symptoms of acute respiratory tract
infection in immune-compromised persons, particularly adults, more
particularly pregnant women and the elderly, the method comprising
administering, to the person, an effective dose of 2'-FL and LNnT
and/or LNT.
[0015] In certain embodiments, the dose of 2'-FL and LNnT and/or
LNT is administered prophylactically to the person on a daily basis
to the immune-compromised person. In some embodiments, the dose is
administered prophylactically in periods of higher risk, for
example during winter.
[0016] In some embodiments, a daily dose of at least 0.5 g of 2'-FL
and LNnT and/or LNT, more advantageously at least 1 g of 2'-FL and
LNnT and/or LNT, for example about 1.5 g or more of 2'-FL and LNnT
and/or LNT. More advantageously, the person is administered the
2'-FL and LNnT and/or LNT in a single daily dose of at least about
2 g of 2'-FL and LNnT and/or LNT per day, even more advantageously
at least 1 3 g of 2'-FL and LNnT and/or LNT.
[0017] In certain embodiments, the abundance of bifidobacteria in
the intestine of the person is increased by the dose of 2'-FL and
LNnT and/or LNT, more advantageously a bifidobacterium of the B.
adolescentis phylogenetic group, especially B. adolescentis and/or
B. pseudocatenulatum.
[0018] In some embodiments, the 2'-FL and LNnT and/or LNT are
effective in reducing the risk of ARI and/or relieving symptoms of
ARI, and in particular reducing the risk and/or relieving symptoms
of bronchitis in immune-compromised persons.
DETAILED DESCRIPTION
[0019] Reference throughout this specification to "one embodiment,"
"an embodiment," or similar language means that a particular
feature, structure, or characteristic described in connection with
the embodiment is included in at least one embodiment of the
present invention. Thus, appearances of the phrases "in one
embodiment," "in an embodiment," and similar language throughout
this specification may, but do not necessarily, all refer to the
same embodiment.
[0020] Furthermore, the described features, structures, or
characteristics of the invention may be combined in any suitable
manner in one or more embodiments. One skilled in the relevant art
will recognize, however, that the invention may be practiced
without one or more of the specific details, or with other methods,
components, materials, and so forth. In other instances, well-known
structures, materials, or procedures are not shown or described in
detail to avoid obscuring aspects of the invention.
[0021] The following terms preferably have the following
meanings:
[0022] "Immune-compromised" means that the immune system of a
person, particularly an adult, is significantly weakened or absent.
Examples of immune-compromised people are HIV and AIDS patients,
patients undergoing chemotherapy or radiation therapy for cancer,
patients having certain cancers and genetic disorders, pregnant
women, and the elderly. Immune-compromised people can also be those
who cannot or should not take pharmaceutical medications against
infections, such as pregnant women.
[0023] "Elderly" means a person of age 60 or above.
[0024] "Adult" means a person above age 13.
[0025] "Preventing AM" means the prevention and the reduction of
frequency and/or occurrence and/or severity and/or duration of AM.
Occurrence is related to the number of any AM. Frequency is related
to the number of the same AM.
[0026] "Relieving symptoms of ARI" means reducing the symptoms of
ARI, and in particular reducing fever, easing the breathing process
and/or diminishing the pain and/or easing sleep.
[0027] "Nutritional composition" means a composition which
nourishes a person. A nutritional composition is usually taken
orally or enterally and usually includes one or more
macronutrients, such as lipids, fats, and proteins, and one or more
micronutrients.
[0028] "Probiotic" means a microbial cell preparation or components
of microbial cells with a beneficial effect on the health or
well-being of the host (Salminen et al. Trends Food Sci. Technol.
10, 107 (1999)).
[0029] "Bifidobacteria of the B. adolescentis phylogenetic group"
is meant a bacterium selected from a group consisting of
Bifidobacterium adolescentis, Bifidobacterium angulatum,
Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum,
Bifidobacterium kashiwanohense, Bifidobacterium dentum and
Bifidobacterium stercoris (Duranti et al. Appl. Environ. Microbiol.
79, 336 (2013), Bottacini et al. Microbial Cell Fact. 13:S4
(2014)).
[0030] The invention is based upon the surprising finding that a
combination of 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose
(LNnT) and/or lacto-N-tetraose (LNT) is effective to protect
immune-compromised persons, particularly adults, against ARI and/or
relieve symptoms of ARI. 2'-FL and LNnT and/or LNT are
oligosaccharides found in human breast milk and safe for long-term
consumption. Therefore, the combination is particularly suitable
for protecting pregnant women and the elderly where reduced drug
intake is advisable.
[0031] The 2'-FL and LNnT and/or LNT can be produced by well-known
processes using microbial fermentation, enzymatic processes,
chemical syntheses, or combinations of these technologies. As
examples, using chemistry, LNnT can be made as described in WO
2011/100980 and WO 2013/044928, LNT can be synthesized as described
in WO 2012/155916 and WO 2013/044928, a mixture of LNT and LNnT can
be made as described in WO 2013/091660, and 2'-FL can be made as
described in WO 2010/115934 and WO 2010/115935.
[0032] To obtain the benefits of preventing ARI and/or relieving
symptoms of ARI, the immune-compromised person is administered a
composition containing, preferably consisting essentially of, 2'-FL
and LNnT and/or LNT. The composition can be in any suitable form,
such as, for example, a nutritional composition, a nutritional
supplement, a drink, a food, a medical device or a pharmaceutical.
The supplement, medical device or pharmaceutical can be in a
galenic form such as a tablet, capsule, pastille, powder in a
sachet or a liquid.
[0033] The composition of the invention is preferably a synthetic
composition. The term "synthetic composition" designates a
composition which is artificially prepared and preferably means a
composition containing at least one compound that is produced ex
vivo chemically and/or biologically, e.g. by means of chemical
reaction, enzymatic reaction or recombinantly. In some embodiments
a synthetic composition of the invention may be, but preferably is
not, identical with a naturally occurring composition. The
synthetic composition of the invention typically comprises 2'-FL
and LNnT and/or LNT, and in some embodiments may comprise one or
more compounds or components other than 2'-FL and LNnT and/or LNT
that may have an effect on bifidobacteria of a human subject
microbiota in vivo, e.g. non-digestible oligosaccharides or
prebiotics. In some embodiments, the synthetic compositions may
comprise one or more nutritionally or pharmaceutically active
components which do not affect adversely the efficacy of the
above-mentioned compounds. According to one embodiment, the
composition of the invention can be co-administered to a person
along with a vaccine, preferably an influenza vaccine. Some
non-limiting embodiments of a synthetic composition of the
invention are also described below.
[0034] Nutritional Compositions
[0035] A suitable nutritional composition can contain a source of
protein, lipids and/or digestible carbohydrates and can be in
powdered, liquid or solid form. The composition can be formulated
to be the sole source of nutrition or as a nutritional
supplement.
[0036] Suitable protein sources include milk proteins, soy protein,
rice protein, pea protein and oat protein, or mixtures of these.
Milk proteins can be in the form of milk protein concentrates, milk
protein isolates, whey protein or casein, or mixtures of these. The
protein can be a whole protein or a hydrolysed protein, either
partially hydrolysed or extensively hydrolysed. The protein can
also be provided in the form of free amino acids. The protein can
comprise about 5% to about 30% of the energy of the nutritional
composition, normally about 10% to 20%.
[0037] The protein source can be a source of glutamine and/or
cysteine. The glutamine source can be a glutamine dipeptide and/or
a glutamine enriched protein. Glutamine can be included due to its
use by enterocytes as an energy source. Cysteine is a major
precursor of glutathione, which is key for the antioxidant defences
of the body.
[0038] Suitable digestible carbohydrates include maltodextrin,
hydrolysed or modified starch or corn starch, glucose polymers,
corn syrup, corn syrup solids, high fructose corn syrup,
rice-derived carbohydrates, pea-derived carbohydrates,
potato-derived carbohydrates, tapioca, sucrose, glucose, fructose,
sucrose, lactose, honey, sugar alcohols (e.g., maltitol,
erythritol, sorbitol), or mixtures of these. Generally, digestible
carbohydrates provide about 35% to about 55% of the energy of the
nutritional composition. Preferably, the nutritional composition is
low in or free from lactose. A particularly suitable digestible
carbohydrate is a low dextrose equivalent (DE) maltodextrin.
[0039] Suitable lipids include medium chain triglycerides (MCT) and
long chain triglycerides (LCT). Suitable sources of long chain
triglycerides are rapeseed oil, sunflower seed oil, palm oil, soy
oil, milk fat, corn oil, high oleic oils, and soy lecithin.
Fractionated coconut oils are a suitable source of medium chain
triglycerides. Generally, the lipids provide about 35% to about 50%
of the energy of the nutritional composition. The lipids can
include polyunsaturated fatty acids such as omega-6 and omega-3
fatty acids. Preferably, polyunsaturated fatty acids provide less
than about 30% of total energy of the lipid source. Decreasing the
levels of polyunsaturated fatty acids is believed to decrease
sensitivity to peroxidation which can be beneficial for persons
having inflammatory conditions.
[0040] The nutritional composition preferably also includes
vitamins and minerals. If the nutritional composition is intended
to be a sole source of nutrition, it preferably includes a complete
vitamin and mineral profile. Examples of vitamins include vitamins
A, B-complex (such as B1, B2, B6 and B12), C, D, E and K, niacin
and acid vitamins such as pantothenic acid, folic acid and biotin.
Examples of minerals include calcium, iron, zinc, magnesium,
iodine, copper, phosphorus, manganese, potassium, chromium,
molybdenum, selenium, nickel, tin, silicon, vanadium and boron. For
example, the composition can contain per daily dose one or more of
the following micronutrients in the ranges given: 300 to 500 mg
calcium, 50 to 100 mg magnesium, 150 to 250 mg phosphorus, 5 to 20
mg iron, 1 to 7 mg zinc, 0.1 to 0.3 mg copper, 50 to 200 .mu.g
iodine, 5 to 15 .mu.g selenium, 1000 to 3000 .mu.g beta carotene,
10 to 80 mg Vitamin C, 1 to 2 mg Vitamin B1, 0.5 to 1.5 mg Vitamin
B6, 0.5 to 2 mg Vitamin B2, 5 to 18 mg niacin, 0.5 to 2.0 .mu.g
Vitamin B12, 100 to 800 .mu.g folic acid, 30 to 70 .mu.g biotin, 1
to 5 .mu.g Vitamin D, 3 to 10 .mu.g Vitamin E.
[0041] The nutritional composition can also include a carotenoid
such as lutein, lycopene, zeaxanthin, and beta-carotene. The total
amount of carotenoid included can vary from about 0.001 .mu.g/ml to
about 10 .mu.g/ml. Lutein can be included in an amount of from
about 0.001 .mu.g/ml to about 10 .mu.g/ml, preferably from about
0.044 .mu.g/ml to about 5 g/ml of lutein. Lycopene can be included
in an amount from about 0.001 .mu.g/ml to about 10 .mu.g/ml,
preferably about 0.0185 mg/ml to about 5 g/ml of lycopene.
Beta-carotene can comprise from about 0.001 .mu.g/ml to about 10
mg/ml, for example about 0.034 .mu.g/ml to about 5 .mu.g/ml of
beta-carotene.
[0042] The nutritional composition can further comprise at least
one probiotic bacterial strain. Suitable probiotic bacterial
strains include Lactobacillus rhamnosus ATCC 53103, Lactobacillus
rhamnosus CGMCC 1.3724, Lactobacillus paracasei CNCM 1-21 16,
Lactobacillus reuteri, Lactobacillus johnsonii CN CM 1-1225,
Streptococcus salivarius DSM 13084, Bifidobacterium lactis CNCM
1-3446, Bifidobacterium longum ATCC BAA-999, Bifidobacterium breve,
Bifidobacterium infantis. Preferably, the nutritional composition
contains from 10e3 to 10e12 cfu of the probiotic bacterial strain
per g of composition on a dry weight basis, more preferably between
10e7 and 10e12 cfu.
[0043] If necessary, the nutritional composition can contain
emulsifiers and stabilisers such as soy, lecithin, citric acid
esters of mono- and di-glycerides, and the like.
[0044] The nutritional composition can also contain other
substances which can have a beneficial effect such as lactoferrin,
nucleotides, nucleosides, and the like.
[0045] For liquid applications, the nutritional composition can be
in the form of a reconstitutable powder, a liquid concentrate, or a
ready-to-use formulation. In liquid form, the nutritional
composition can be fed to a person via a nasogastric tube or by
having the person drink it. Various flavours, fibres and other
additives can also be present.
[0046] The nutritional composition can be prepared in any suitable
manner. For example, it can be prepared by blending together the
protein, the digestible carbohydrate source, and the fat source in
appropriate proportions. If used, the emulsifiers can be included
at this point. The vitamins and minerals can be added at this point
but are usually added later to avoid thermal degradation. Any
lipophilic vitamins, emulsifiers and the like can be dissolved into
the fat source prior to blending. Water, preferably water which has
been subjected to reverse osmosis, can then be mixed in to form a
liquid mixture. The temperature of the water is conveniently in the
range between about 50.degree. C. and about 80.degree. C. to aid
dispersal of the ingredients. Commercially available liquefiers can
be used to form the liquid mixture. The 2'-FL and LNnT and/or LNT
are added at this stage if the final product will be in liquid
form. If the final product is to be a powder, the oligosaccharides
can be added at this stage if desired. The liquid mixture is then
homogenised, for example in two stages.
[0047] The liquid mixture can then be thermally treated to reduce
bacterial loads, by rapidly heating the liquid mixture to a
temperature in the range between about 80.degree. C. and about
150.degree. C. for a duration between about 5 seconds and about 5
minutes, for example. This can be carried out by means of steam
injection, an autoclave or a heat exchanger, for example a plate
heat exchanger.
[0048] Then, the liquid mixture can be cooled to between about
60.degree. C. and about 85.degree. C. for example by flash cooling.
The liquid mixture can then be again homogenised, for example in
two stages between about 10 MPa and about 30 MPa in the first stage
and between about 2 MPa and about 10 MPa in the second stage. The
homogenised mixture can then be further cooled to add any heat
sensitive components, such as vitamins and minerals. The pH and
solids content of the homogenised mixture are conveniently adjusted
at this point.
[0049] The homogenised mixture is transferred to a suitable drying
apparatus such as a spray dryer or freeze dryer and converted to
powder. The powder should have a moisture content of less than
about 5% by weight. The 2'-FL and LNnT and/or LNT can be added at
this stage by dry-mixing, or by blending them in a syrup form of
crystals, along with the probiotic bacterial strain(s) if used, and
spray-dried (or freeze-dried).
[0050] If a liquid composition is preferred, the homogenised
mixture can be sterilised then aseptically filled into suitable
containers or can be first filled into the containers and then
retorted.
[0051] When the nutritional product is a ready-to-feed nutritional
liquid, the total concentration of 2'-FL and LNnT and/or LNT in the
liquid, by weight of the liquid, is from about 0.02% to about 2.0%,
including from about 0.03% to about 1.5%, including from about
0.04% to about 1.0%. When the nutritional product is a concentrated
nutritional liquid, the total concentration of HMOs in the liquid,
by weight of the liquid, is from about 0.04% to about 4.0%,
including from about 0.06% to about 3.0%, including from about
0.08% to about 2.0%.
[0052] The nutritional composition can also be formulated into
solid and semi-solid forms such as cereal bars, yoghurts and the
like.
[0053] Galenic Forms
[0054] The synthetic composition can also be in a galenic form such
as a capsule, tablet or sachet. For example, the composition can be
in a tablet form comprising the HMOs, and one or more additional
components to aid formulation and administration, such as diluents,
excipients, antioxidants, lubricants, colorants, binders,
disintegrants, and the like.
[0055] Suitable diluents, excipients, lubricants, colorants,
binders, and disintegrants include polyethylene, polyvinyl
chloride, ethyl cellulose, acrylate polymers and their copolymers,
hydroxyethyl-cellulose, hydroxypropylmethyl-cellulose (HPMC),
sodium carboxymethylcellulose, polyhydroxyethylmethylacrylate
(PHEMA), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP),
polyethylene oxide (PEO), or polyacrylamide (PA), carrageenan,
sodium alginate, polycarbophil, polyacrylic acid, tragacanth,
methyl cellulose, pectin, natural gums, xanthan gum, guar gum,
karaya gum, hypromellose, magnesium stearate, microcrystalline
cellulose, and colloidal silicon dioxide. Suitable antioxidants are
vitamin A, carotenoids, vitamin C, vitamin E, selenium, flavonoids,
polyphenols, lycopene, lutein, lignan, coenzyme Q10 ("CoQlO") and
glutathione. The unit dosage forms, especially those in sachet
form, can also include various nutrients including
macronutrients.
[0056] The galenic forms can fall within any suitable regulatory
framework, for example they can be nutritional supplements, foods
for special medical purposes/medical foods, medical devices or
drugs.
[0057] Administration Dosing
[0058] For preventing acute respiratory tract infections (ARI)
and/or relieving symptoms of ARI in immune-compromised person,
preferably an adult, the amount of 2'-fucosyllactose and
lacto-N-neotetraose and/or lacto-N-tetraose required to be
administered to the adult will vary depending upon factors such as
the severity of the immune deficiency, the age of the person, the
form of the composition, and other medications being administered
to the person. However, the required amount can be readily set by a
medical practitioner and would generally be in the range from about
200 mg to about 20 g per day, in certain embodiments from about 300
mg to about 15 g per day, from about 400 mg to about 10 g per day,
in certain embodiments from about 500 mg to about 10 g per day, in
certain embodiments from about 1 g to about 10 g per day. An
appropriate dose can be determined based on several factors,
including, for example, the body weight and/or condition of the
person being treated, the severity of the immune deficiency, other
ailments and/or diseases of the person, and the manner of
administration. Appropriate dose regimes can be determined by
methods known to those skilled in the art.
[0059] The composition can be co-administered to a person along
with a vaccine. Co-administration includes the administration of
the composition from three months before the vaccination period to
three months after the vaccination period. Suitable vaccines
include a vaccine against influenza.
Example 1
[0060] An aged mouse model is used to assess the HMO impact on the
immune system of an immune-compromised individual. Ageing is
associated with a noted decline in cell-mediated immune response
concomitant with an increased humoral immune dysfunction. Ageing is
furthermore often associated with a status of low-grade
inflammation and of increased oxidative stress. Consequently, many
elderly are at increased risk of infectious and non-infectious
diseases that contribute to morbidity and mortality.
[0061] Specific pathogen-free male C57BL/6J mice (4-weeks old) are
housed under conventional conditions (12 hours light/dark cycle,
temperature 22.degree. C., humidity 56%) and receive water and a
semisynthetic diet ad libitum. Until the age of 5 months, the mice
are maintained in groups of 5 per cage and then they are
individually caged. At 19 months after birth, the mice are
randomised into 3 groups of 10 animals and are fed either with a
control semi-synthetic diet (control group, n=10) or one of two
semi-synthetic diets supplemented either with 2'-FL and LNnT in a
2:1 ratio (1.2 weight % of 2'-FL and LNnT) (treatment group 1,
n=10) or with 2'-FL and LNnT in a 4:1 ratio (1.2 weight-% of 2'-FL
and LNnT) (treatment group 2, n=10). During the 7 weeks of trial,
all mice are allowed to drink and eat ad libitum.
[0062] On day 15 of the trial, the mice are immunized by
subcutaneous injection (100 .mu.l) of an inert antigen Keyhole
Limpet Haemocyanin (KLH) at 100 .mu.g in 1% Alum. Seven days after
immunization, Delayed-Type Hypersensitivity (DTH) responses are
elicited by injecting the recall-antigen KLH (10 .mu.l of 0.5
.mu.g/ml) into each mouse's right ear. The left ears are injected
with vehicle alone (saline=PBS) and serve as internal controls for
each animal. At 24 hours post-elicitation, and during the following
5 days, both the non-elicited (left ear) and the elicited ears
(right ear) are measured. DTH responses are expressed as the
magnitude of ear swelling, i.e. the change in ear thickness using
the following formula: .DELTA. in ear thickness=[elicited ear
(right) ear sickness-non-elicited (left) ear sickness], where
.DELTA. in ear thickness=[post-elicitation-pre-elicitation ear
thickness].
[0063] Mice are euthanized on day 42 of trial. The expression of
genes involved in inflammatory processes are determined in the
liver of the mice. At the autopsy, the liver is removed and a piece
is immediately frozen in liquid nitrogen and then stored at
-80.degree. C. until further analysis.
[0064] Liver samples are transferred into 1 ml of RNA lysis buffer
and homogenized using a Ribolyzer. RNA is extracted using a
commercially available kit. RNA is quantified a Ribogreen RNA
Quantitation Kit, and RNA quality is assayed using Agilent RNA 6000
Nano LabChip Kit. Total RNA (2 .mu.g) is reverse transcribed using
Multischbe reverse transcriptase following manufacturer's
instructions. Custom-made low density arrays (LDA) with TaqMan
probes are used. Gene expression is calculated using the relative
quantification method .DELTA..DELTA.Ct method with SDS 2.2.2
software (Applied Biosystems). Data are analysed by means+/-SEM and
the Student's T test (unpaired) or two-way ANOVA when appropriate.
Probability values of less than 5% were considered as
significant.
[0065] For both treatment groups, dietary supplementation with
2'-FL and LNnT (1.2 weight % incorporated in the diet) improves
cell-mediated immune response in aged mice. This supplementation
alleviates age-related low-grade inflammation and oxidative damage,
as observed by increased expression of genes encoding for an
anti-inflammatory molecule (TGF-.beta.2) or for molecules involved
in the defence against oxidative damage (HO-1 and SOD2).
Example 2
[0066] A total of 30 of elderly male and female persons are
recruited to participate in the study. The persons are residents of
long-term care facilities for the elderly. After a screening visit
and run-in period of 1-2 weeks, the persons are selected. The
selected individuals are randomized into two groups, each of 15
individuals, with one group consuming the treatment product and one
group the placebo product for 12 weeks. The treatment product
contains 2 grams of a combination of 2'-FL, and LNnT in a 2:1 mass
ratio. The control product contains 2 grams glucose. Both products
are in powder form in a unit dosage container.
[0067] The selected individuals are eligible to participate if they
are able and willing to understand and comply with the study
procedures. Individuals are excluded if: they have: participated in
a clinical study one month prior to screening visit; they have
abnormal results in the screening tests which are clinically
relevant for study participation; they are suffering for a severe
disease; they have used highly dosed probiotic supplements (yoghurt
allowed) for 3 months prior to the study; they have consumed
antibiotic drugs 3 months prior to the study; they have consumed on
a regular basis any medication that might interfere with symptom
evaluation 2 weeks prior to the study; or they are pregnant or
lactating.
[0068] At the screening visit, medical history and concomitant
medication is registered and a blood sample for safety analyses is
collected from each selected individual. A faecal sample kit is
distributed. The selected individuals are instructed to keep their
samples in the freezer until the next visit.
[0069] At the second visit, eligibility criteria are checked and
eligible subjects are randomised to the two arms in the trial. The
faecal samples are collected and equipment for new samples are
distributed. Blood samples are collected for biomarker studies.
[0070] The serum from the blood samples is transferred to cryotubes
and stored at -80.degree. C. The blood is analysed for biomarkers
of safety and inflammatory response.
[0071] The faecal samples are stored at -80.degree. C. until
analysis. Faecal samples are subjected to 16 S RNA sequencing
analysis.
[0072] The study runs for 12 weeks over the winter months with the
subjects consuming either a placebo or a treatment product daily.
Subjects are instructed to consume the products in the morning with
breakfast.
[0073] At the end of the study, each subject has an exit visit with
the medical team. Faecal samples and blood samples are collected
and analysed as before.
[0074] The treatment subjects have reduced incidence of lower
respiratory tract infections, particularly bronchitis. Further,
incidences of fever and antibiotic use are lower in the treatment
group subjects. Also, the treatment group subjects who are
infected, have less acute symptoms and recover more quickly. The
blood biomarker analysis indicates that the treatment subjects have
improved immune status. The faecal analysis indicates that the
treatment subjects have improved levels of beneficial
bifidobacteria, in particular a bifidobacterium of the B.
adolescentis phylogenetic group, especially Bifidobacterium
adolescentis and/or Bifidobacterium pseudocatenulatum.
Example 3--Nutritional Composition
[0075] A ready to feed nutritional composition is prepared from
water, maltodextrin, whey protein (from cows milk), medium chain
triglycerides (from coconut and/or palm kernel oil), cornstarch,
soybean oil, soy lecithin, 2'-FL, LNnT, magnesium chloride, calcium
phosphate, guar gum, sodium ascorbate, potassium citrate, sodium
phosphate, calcium citrate, choline chloride, potassium chloride,
sodium citrate, magnesium oxide, taurine, L-carnitine,
alpha-tocopheryl acetate, zinc sulphate, ferrous sulphate,
niacinamide, calcium pantothenate, vitamin A palmitate, citric
acid, manganese sulphate, pyridoxine hydrochloride, vitamin D3,
copper sulphate, thiamine mononitrate, riboflavin, beta carotene,
folic acid, biotin, potassium iodide, chromium chloride, sodium
selenate, sodium molybdate, phytonadione, vitamin B12.
[0076] The composition has a calorific density of 1.0 kcal/ml with
a caloric distribution (% of kcal) as follows: protein: 16%,
carbohydrate: 51%, fat: 33%. The protein source has an NPC:N ratio
of 131:1. The MCT:LCT ratio is 70:30 and the n6:n3 ratio is 7.4:1.
The osmolality (mOsm/kg water) is 270 when unflavoured. The
composition contains 85% water and 1500 ml meets 100% of the RDI
for 22 key micronutrients.
Example 4 Tablet Composition
[0077] A tablet is prepared from 2'-FL, LNnT, hydroxypropyl
methylcellulose, sodium alginate, gum, microcrystalline cellulose,
colloidal silicon dioxide, and magnesium stearate. All raw
materials except the magnesium stearate are placed into a high
shear granulator and premixed. Water is sprayed onto the premix
while continuing to mix at 300 rpm. The granulate is transferred to
a fluidised bed drier and dried at 75.degree. C. The dried powder
is sieved and sized using a mill. The resulting powder is then
lubricated with magnesium stearate and pressed into tablets. The
tablets each contain 325 mg of 2'-FL and LNnT. The tablets each
have a weight of 750 mg.
Example 5 Capsule Composition
[0078] A capsule is prepared by filling about 1 g of 2'-FL and LNnT
into a 000 gelatine capsule using a filing machine. The capsules
are then closed. The 2'-FL and LNnT are in free flowing, powder
form.
Example 6 Capsule Composition
[0079] A capsule is prepared by filling about 1 g of 2'-FL and LNT
into a 000 gelatine capsule using a filing machine. The capsules
are then closed. The 2'-FL and LNnT are in free flowing, powder
form.
* * * * *