U.S. patent application number 16/211743 was filed with the patent office on 2019-10-10 for bifidobacteria for treating diabetes and related conditions.
The applicant listed for this patent is DUPONT NUTRITION BIOSCIENCES APS. Invention is credited to REMY BURCELIN, DIDIER CARCANO, SAMPO LAHTINEN.
Application Number | 20190307815 16/211743 |
Document ID | / |
Family ID | 42989263 |
Filed Date | 2019-10-10 |
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United States Patent
Application |
20190307815 |
Kind Code |
A1 |
BURCELIN; REMY ; et
al. |
October 10, 2019 |
BIFIDOBACTERIA FOR TREATING DIABETES AND RELATED CONDITIONS
Abstract
This invention relates to new uses of Bifidobacteria
(particularly, although not exclusively, probiotic Bifidobacteria),
and to food products, feed products, dietary supplements and
pharmaceutical formulations containing them. The bacteria are
suitable for the treatment of diabetes (particularly Type 2
diabetes), obesity and related conditions, metabolic syndrome,
insulin resistance, and impaired glucose metabolism and
consequences thereof, lowering tissue inflammation, treating
hepatitis, myositis and cardiovascular conditions.
Inventors: |
BURCELIN; REMY; (ESCALQUENS,
FR) ; CARCANO; DIDIER; (PARIS, FR) ; LAHTINEN;
SAMPO; (LOHJA, FI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
DUPONT NUTRITION BIOSCIENCES APS |
COPENHAGEN |
|
DK |
|
|
Family ID: |
42989263 |
Appl. No.: |
16/211743 |
Filed: |
December 6, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15452240 |
Mar 7, 2017 |
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16211743 |
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13377325 |
Jan 13, 2012 |
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PCT/IB10/52757 |
Jun 18, 2010 |
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15452240 |
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61218563 |
Jun 19, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
Y02A 50/30 20180101;
A61K 2035/115 20130101; A61K 35/747 20130101; A61K 31/715 20130101;
A23Y 2300/21 20130101; A23V 2002/00 20130101; Y02A 50/463 20180101;
A61P 3/04 20180101; A61K 45/06 20130101; A61K 2300/00 20130101;
A61K 31/155 20130101; A23L 33/135 20160801; A61P 5/50 20180101;
A61K 35/745 20130101; A23C 9/123 20130101; A61K 31/155 20130101;
A61K 2300/00 20130101; A61K 35/747 20130101; A61K 2300/00 20130101;
A61K 35/745 20130101; A61K 2300/00 20130101; A23V 2002/00 20130101;
A23V 2200/328 20130101; A23V 2200/332 20130101; A23V 2200/324
20130101; A23V 2200/326 20130101 |
International
Class: |
A61K 35/745 20060101
A61K035/745; A61K 31/715 20060101 A61K031/715; A61K 31/155 20060101
A61K031/155; A61K 35/747 20060101 A61K035/747; A61K 45/06 20060101
A61K045/06; A23C 9/123 20060101 A23C009/123; A61P 5/50 20060101
A61P005/50; A61P 3/04 20060101 A61P003/04; A23L 33/135 20060101
A23L033/135 |
Claims
1-44 (canceled)
45. A method of treating a disease or condition in a mammal,
wherein: the method comprises administering to a mammal in need of
such treatment a combination of (i) a bacterium of the genus
Bifidobacterium and (ii) an antidiabetic drug; and the disease or
condition is selected from: (a) diabetes; (b) metabolic syndrome;
(c) impaired glucose tolerance; (d) reduced insulin sensitivity;
(e) reduced fed insulin secretion; (f) elevated fasted insulin
secretion; (g) obesity; (h) weight gain; (i) elevated body fat
mass; and (j) elevated mesenteric fat mass.
46. The method of claim 45, wherein: the method comprises treating
diabetes, and the diabetes is Type 2 diabetes.
47-51 (canceled)
52. The method of claim 45, wherein the mammal in need of the
treatment ingests a high-fat diet.
53. The method of claim 45, wherein the Bifidobacterium is a
probiotic Bifidobacterium.
54. The method of claim 45, wherein the Bifidobacterium is selected
from the species Bifidobacterium lactis, Bifidobacterium bifidium,
Bifidobacterium longum, Bifidobacterium animalis, Bifidobacterium
breve, Bifidobacterium infantis, Bifidobacterium catenulatum,
Bifidobacterium pseudocatenulatum, Bifidobacterium adolescentis,
and Bifidobacterium angulatum.
55. The method of claim 54, wherein the Bifidobacterium is of the
species Bifidobacterium animalis.
56. The method of claim 55, wherein the Bifidobacterium is of the
species Bifidobacterium animalis subsp. lactis.
57. The method of claim 45, wherein the Bifidobacterium is of the
species Bifidobacterium animalis subsp. lactis strain 420
(B420).
58. The method of claim 45, wherein the Bifidobacterium and
antidiabetic drug are administered in combination with an
additional bacterium of the genus Lactobacillus.
59. The method of claim 58, wherein the additional bacterium is of
the species Lactobacillus acidophilus.
60. The method of claim 59, wherein the additional bacterium is
Lactobacillus acidophilus strain NCFM (ATCC PTA-4797).
61. The method of claim 45, wherein the Bifidobacterium and
antidiabetic drug are administered in combination with a
prebiotic.
62. The method of claim 61, wherein the prebiotic is
polydextrose.
63. (canceled)
64. The method of claim 45, wherein the antidiabetic drug is
selected from a biguanide, a sulfonylurea, an alpha-glucosidase
inhibitor, a thiazolidinedione, a meglitinide, a dipeptidyl
peptidase-4 (DPP-4) inhibitor, a glucagon-like peptide-1 analog, an
amylin analog, a fast acting insulin analog, a long acting insulin
analog, a dual PPAR agonist and an SGLT2 inhibitor.
65. The method of claim 45, wherein the antidiabetic drug is a
biguanide.
66. The method of claim 45, wherein the antidiabetic drug is
metformin.
67. The method of claim 45, wherein the antidiabetic drug is
selected from metformin, carbutamide, chlorpropamide,
glibenclamide, gliclazide, glimepiride, glipizide, gliquidone,
tolazamide, tolbutamide), acarbose, miglitol, voglibose,
pioglitazone, rivoglitazone, rosiglitazone, nateglinide,
repaglinide, mitiglinide, alogliptin, saxagliptin, sitagliptin,
vildagliptin, exenatide, liraglutide, albiglutide, pramlintide,
insulin lispro, insulin aspart, insulin glulisine, insulin
glargine, insulin detemir, aleglitazar, dapagliflozin,
remogliflozin and sergliflozin.
68. The method of claim 45, wherein the antidiabetic drug is
selected from alogliptin, saxagliptin, sitagliptin and
vildagliptin.
69. The method of claim 57, wherein: the method comprises treating
diabetes, and the diabetes is Type 2 diabetes.
70. The method of claim 57, wherein the mammal in need of the
treatment ingests a high-fat diet.
71. The method of claim 57, wherein the Bifidobacterium and
antidiabetic drug are administered in combination with an
additional bacterium of the genus Lactobacillus.
72. The method of claim 71, wherein the additional bacterium is of
the species Lactobacillus acidophilus.
73. The method of claim 71, wherein the additional bacterium is
Lactobacillus acidophilus strain NCFM (ATCC PTA-4797).
74. The method of claim 57, wherein the antidiabetic drug is
selected from metformin, carbutamide, chlorpropamide,
glibenclamide, gliclazide, glimepiride, glipizide, gliquidone,
tolazamide, tolbutamide), acarbose, miglitol, voglibose,
pioglitazone, rivoglitazone, rosiglitazone, nateglinide,
repaglinide, mitiglinide, alogliptin, saxagliptin, sitagliptin,
vildagliptin, exenatide, liraglutide, albiglutide, pramlintide,
insulin lispro, insulin aspart, insulin glulisine, insulin
glargine, insulin detemir, aleglitazar, dapagliflozin,
remogliflozin and sergliflozin.
75. The method of claim 57, wherein the antidiabetic drug is
selected from a biguanide, a sulfonylurea, an alpha-glucosidase
inhibitor, a thiazolidinedione, a meglitinide, a dipeptidyl
peptidase-4 (DPP-4) inhibitor, a glucagon-like peptide-1 analog, an
amylin, a fast acting insulin analog, a long acting insulin analog,
a dual PPAR agonist and an SGLT2 inhibitor.
76. The method of claim 57, wherein the antidiabetic drug is a
biguanide.
77. The method of claim 57, wherein the antidiabetic drug is
metformin.
78. The method of claim 57, wherein the antidiabetic drug is a
dipeptidyl peptidase-4 (DPP-4) inhibitor.
79. The method of claim 64, wherein the antidiabetic drug is a
dipeptidyl peptidase-4 (DPP-4) inhibitor or a biguanide.
80. The method of claim 79, wherein: the method comprises treating
diabetes, and the diabetes is Type 2 diabetes.
81. The method of claim 79, wherein the mammal in need of the
treatment ingests a high-fat diet.
82. The method of claim 79, wherein the Bifidobacterium is selected
from the species Bifidobacterium lactis, Bifidobacterium bifidium,
Bifidobacterium longum, Bifidobacterium animalis, Bifidobacterium
breve, Bifidobacterium infantis, Bifidobacterium catenulatum,
Bifidobacterium pseudocatenulatum, Bifidobacterium adolescentis,
and Bifidobacterium angulatum.
83. The method of claim 79, wherein the Bifidobacterium is of the
species Bifidobacterium animalis.
84. The method of claim 79, wherein the Bifidobacterium is of the
species Bifidobacterium animalis subsp. lactis strain 420
(B420).
85. The method of claim 79, wherein the Bifidobacterium and
antidiabetic drug are administered in combination with an
additional bacterium of the genus Lactobacillus.
Description
FIELD OF THE INVENTION
[0001] This invention relates to new uses of Bifidobacteria
(particularly, although not exclusively, probiotic Bifidobacteria),
and to food products, feed products, dietary supplements and
pharmaceutical formulations containing them.
DESCRIPTION OF THE PRIOR ART
[0002] Diabetes mellitus, often referred to simply as diabetes, is
a condition characterized by disordered metabolism and abnormally
high blood sugar (hyperglycaemia) resulting from insufficient
levels and/or action of the hormone insulin. The characteristic
symptoms are excessive urine production (polyuria) due to high
blood glucose levels, excessive thirst and increased fluid intake
(polydipsia) attempting to compensate for increased urination,
blurred vision due to high blood glucose effects on the eye's
optics, unexplained weight loss, and lethargy. These symptoms are
likely to be less apparent if the blood sugar is only mildly
elevated.
[0003] The World Health Organisation recognises three main forms of
diabetes mellitus: type 1, type 2, and gestational diabetes
(occurring during pregnancy), which have different causes and
population distributions. While, ultimately, all forms are due to
the beta cells of the pancreas being unable to produce sufficient
insulin to prevent hyperglycemia, the causes are different. Type 1
diabetes is usually due to autoimmune destruction of the pancreatic
beta cells. Type 2 diabetes is characterized by insulin resistance
in target tissues. This causes a need for abnormally high amounts
of insulin and diabetes develops when the beta cells cannot meet
this demand. Gestational diabetes is similar to type 2 diabetes in
that it involves insulin resistance; the hormones of pregnancy can
cause insulin resistance in women genetically predisposed to
developing this condition.
[0004] Gestational diabetes typically resolves with delivery of the
child: however, types 1 and 2 diabetes are chronic conditions. All
types have been treatable since insulin became medically available
in 1921. Type 1 diabetes, in which insulin is not secreted by the
pancreas, is directly treatable only with injected insulin,
although dietary and other lifestyle adjustments re part of
management. Type 2 may be managed with a combination of dietary
treatment, tablets and injections and, frequently, insulin
supplementation.
[0005] Diabetes can cause many complications. Acute complications
(hypoglycemia, ketoacidosis or nonketotic hyperosmolar coma) may
occur if the disease is not adequately controlled. Serious
long-term complications include cardiovascular disease (doubled
risk), chronic renal failure, retinal damage (which can lead to
blindness), nerve damage (of several kinds), and microvascular
damage, which may cause impotence and poor healing. Poor healing of
wounds, particularly of the feet, can lead to gangrene, which may
require amputation. Adequate treatment of diabetes, as well as
increased emphasis on blood pressure control and lifestyle factors
(such as not smoking and keeping a healthy body weight), may
improve the risk profile of most aforementioned complications. In
the developed world, diabetes is the most significant cause of
adult blindness in the non-elderly and the leading cause of
non-traumatic amputation in adults, and diabetic nephropathy is the
main illness requiring renal dialysis in the United States.
[0006] Diabetes mellitus is currently a chronic disease, without a
cure, and medical emphasis must necessarily be on managing/avoiding
possible short-term as well as long-term diabetes-related problems.
There is an exceptionally important role for patient education,
dietetic support, sensible exercise, self glucose monitoring, with
the goal of keeping both short-term blood glucose levels, and long
term levels as well, within acceptable bounds. Careful control is
needed to reduce the risk of long term complications. This is
theoretically achievable with combinations of diet, exercise and
weight loss (type 2), various oral diabetic drugs (type 2 only),
and insulin use (type 1 and increasingly for type 2 not responding
to oral medications). In addition, given the associated higher
risks of cardiovascular disease, lifestyle modifications should be
undertaken to control blood pressure and cholesterol by exercising
more, smoking cessation, consuming an appropriate diet, wearing
diabetic socks, and if necessary, taking any of several drugs to
reduce pressure.
[0007] Oral antidiabetic drugs and insulin analogs currently on the
market or undergoing clinical trials include biguanides (such as
metformin), sulfonylureas (such as carbutamide, chlorpropamide,
glibenclamide (Glyburide), gliclazide, glimepiride, glipizide,
gliquidone, tolazamide or tolbutamide), alpha-glucosidase
inhibitors (such as acarbose, miglitol or voglibose),
thiazolidinediones (TZD) (such as pioglitazone, rivoglitazone or
rosiglitazone), meglitinides (such as nateglinide, repaglinide or
mitiglinide), dipeptidyl peptidase-4 (DPP-4) inhibitors (such as
alogliptin, saxagliptin, sitagliptin car vildagliptin),
glucagon-like peptide-1 analogs (such as exenatide, liraglutide, or
albiglutide), amylin analogs (such as pramlintide), fast acting
insulin analogs (such as insulin lispro, insulin aspart and insulin
glulisine), long acting insulin analogs (such as insulin glargine,
insulin detemir), dual PPAR agonists (such as aleglitazar) and
SGLT2 inhibitors (such as dapagliflozin, remogliflozin and
sergliflozin).
[0008] Type 2 diabetes is often associated with obesity. The body
mass index (BMI) (calculated as weight in kilograms divided by the
square of height in metres) is the most commonly accepted
measurement for overweight and/or obesity. A BMI exceeding 25 is
considered overweight. Obesity is defined as a BMI of 30 or more,
with a BMI of 35 or more considered as serious comorbidity obesity
and a BMI of 40 or more considered morbid obesity. Mortality is
increased in obesity, with a BMI of over 32 being associated with a
doubled risk of death. There are alterations in the body's response
to insulin (insulin resistance), a proinflammatory state and an
increased tendency to thrombosis (prothrombotic state).
[0009] Central obesity (male-type or waist-predominant obesity,
characterised by a high waist-hip ratio), is a particularly
important risk factor for diabetes and metabolic syndrome, the
clustering of a number of diseases and risk factors that heavily
predispose for cardiovascular disease. These are diabetes mellitus
type 2, high blood pressure, high blood cholesterol, and
triglyceride levels (combined hyperlipidemia).
[0010] The use of microorganisms in treating obesity, diabetes and
diabetes-related conditions is in general known in the art. For
example, WO 2007/043933 describes the use of probiotic bacteria for
the manufacture of food and feed products, dietary supplements, for
controlling weight gain, preventing obesity, increasing satiety,
prolonging satiation, reducing food intake, reducing fat
deposition, improving energy metabolism, enhancing insulin
sensitivity, treating obesity and treating insulin
insensitivity.
[0011] WO 2009/024429 describes the use of a primary composition
comprising an agent that reduces the amount of proteobacteria, in
particular enterobacteria and/or deferribacteres in the gut for the
treatment or prevention of metabolic disorders, to support and/or
to support weight management.
[0012] WO 2009/004076 describes the use of probiotic bacteria for
normalising plasma glucose concentrations, improving insulin
sensitivity, and reducing the risk of development in pregnant
women, and preventing gestational diabetes.
[0013] WO 2009/021824 describes the use of probiotic bacteria, in
particular Lactobacillus rhamnosus, to treat obesity, treat
metabolic disorders, and support weight loss and/or weight
maintenance.
[0014] WO 2008/016214 describes a probiotic lactic acid bacterium
of the strain Lactobacillus gasseri BNR17 and its use in the
inhibition of weight gain.
[0015] WO 02/38165 describes use of a strain of Lactobacillus (in
particular, Lactobacillus plantarum) in reducing the risk factors
involved in the metabolic syndrome.
[0016] US 2002/0037577 describes the use of microorganisms, such as
Lactobacilli, for the treatment or prevention of obesity or
diabetes mellitus by reduction of the amount of monosaccharide or
disaccharide which may be absorbed into the body, by converting
such compounds into polymeric materials which cannot be absorbed by
the intestine.
[0017] Lee et al., J. Appl. Microbiol. 2007, 103, 1140-1146,
describes the anti-obesity activity of trans-10, cis-12-conjugated
linoleic acid (CLA)-producing bacterium of the strain Lactobacillus
plantarum PL62 in mice.
[0018] Li et al., Hepatology, 2003, 37(2), 343-350, describe the
use of probiotics and anti-TNF antibodies in a mouse model for
non-alcoholic fatty liver disease.
SUMMARY OF THE INVENTION
[0019] In one aspect, the invention comprises use of a bacterium of
the genus Bifidobacterium or a mixture thereof in the manufacture
of a food product, dietary supplement or medicament for treating
diabetes (preferably but not exclusively Type 2 diabetes) in a
mammal.
[0020] In another aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating impaired glucose tolerance in a mammal.
[0021] In a further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
normalising insulin sensitivity in a mammal.
[0022] In a yet further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
increasing fed insulin secretion in a mammal.
[0023] In a still further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
decreasing fasted insulin secretion in a mammal.
[0024] In an additional aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
improving glucose tolerance in a mammal.
[0025] In another aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating obesity, controlling weight gain and/or inducing weight
loss in a mammal.
[0026] In a further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
lowering body fat mass in a mammal.
[0027] In a yet further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
lowering mesenteric fat mass in a mammal.
[0028] In a still further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
lowering tissue inflammation (particularly, although not
exclusively, muscle tissue inflammation, liver tissue inflammation
and/or adipose tissue inflammation) in a mammal.
[0029] In a still further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating hepatitis in a mammal.
[0030] In a yet further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating myositis in a mammal.
[0031] In a still further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating cardiovascular disease in a mammal.
[0032] In a yet further aspect, the invention comprises use of a
bacterium of the genus Bifidobacterium or a mixture thereof in the
manufacture of a food product, dietary supplement or medicament for
treating metabolic syndrome in a mammal.
[0033] In another aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in treating
diabetes (particularly, although not exclusively, Type 2 diabetes)
in a mammal.
[0034] In a further aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in treating
impaired glucose tolerance in a mammal.
[0035] In a yet further aspect, the invention comprises a bacterium
of the genus Bifidobacterium or a mixture thereof for use in
normalising insulin sensitivity in a mammal.
[0036] In a still further aspect, the invention comprises a
bacterium of the genus Bifidobacterium or a mixture thereof for use
in increasing fed insulin secretion in a mammal.
[0037] In another aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in
decreasing fasted insulin secretion in a mammal.
[0038] In a further aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in improving
glucose tolerance in a mammal.
[0039] In a yet further aspect, the invention comprises a bacterium
of the genus Bifidobacterium or a mixture thereof for use in
treating obesity, controlling weight gain and/or inducing weight
loss in a mammal.
[0040] In another aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in lowering
body fat mass in a mammal.
[0041] In a further aspect, the invention comprises a bacterium of
the genus Bifidobacterium or a mixture thereof for use in lowering
mesenteric fat mass in a mammal.
[0042] In a yet further aspect, the invention comprises a bacterium
of the genus Bifidobacterium or a mixture thereof for use in
lowering tissue inflammation (particularly, although not
exclusively, muscle tissue inflammation, liver tissue inflammation
and/or adipose tissue inflammation) in a mammal.
[0043] In a still further aspect, the invention comprises a
bacterium of the genus Bifidobacterium or a mixture thereof for use
in treating hepatitis in a mammal.
[0044] In a yet further aspect, the invention comprises a bacterium
of the genus Bifidobacterium or a mixture thereof for use in
treating myositis in a mammal.
[0045] In a still further aspect, the invention comprises a
bacterium of the genus Bifidobacterium or a mixture thereof for use
in treating cardiovascular disease in a mammal.
[0046] In a still further aspect, the invention comprises a
bacterium of the genus Bifidobacterium or a mixture thereof for use
in treating metabolic syndrome in a mammal.
[0047] In another aspect, the invention comprises a method of
treating diabetes (particularly although not exclusively Type 2
diabetes) in a mammal, comprising administering to a mammal in need
of such treatment a bacterium of the genus Bifidobacterium or a
mixture thereof.
[0048] In a further aspect, the invention comprises a method of
treating impaired glucose tolerance in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0049] In a yet further aspect, the invention comprises a method of
normalising insulin sensitivity in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0050] In a still further aspect, the invention comprises a method
of increasing fed insulin secretion in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0051] In another aspect, the invention comprises a method of
decreasing fasted insulin secretion in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0052] In a further aspect, the invention comprises a method of
improving glucose tolerance in a mammal, comprising administering
to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0053] In a yet further aspect, the invention comprises a method of
treating obesity, controlling weight gain and/or inducing weight
loss in a mammal, comprising administering to a mammal in need of
such treatment a bacterium of the genus Bifidobacterium or a
mixture thereof.
[0054] In a still further aspect, the invention comprises a method
of lowering body fat mass in a mammal, comprising administering to
a mammal in need thereof a bacterium of the genus Bifidobacterium
or a mixture thereof.
[0055] In another aspect, the invention comprises a method of
lowering mesenteric fat mass in a mammal, comprising administering
to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0056] In a further aspect, the invention comprises a method of
lowering tissue inflammation (particularly, although not
exclusively, muscle tissue inflammation, liver tissue inflammation
and/or adipose tissue inflammation) in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0057] In a yet further aspect, the invention comprises a method of
treating hepatitis in a mammal, comprising administering to a
mammal in need thereof a bacterium of the genus Bifidobacterium or
a mixture thereof.
[0058] In a still further aspect, the invention comprises a method
of treating myositis in a mammal, comprising administering to a
mammal in need thereof a bacterium of the genus Bifidobacterium or
a mixture thereof.
[0059] In a yet further aspect, the invention comprises a method of
treating cardiovascular disease in a mammal, comprising
administering to a mammal in need thereof a bacterium of the genus
Bifidobacterium or a mixture thereof.
[0060] In a still further aspect, the invention comprises a method
of treating metabolic syndrome in a mammal, comprising
administering to a mammal in need of such treatment a bacterium of
the genus Bifidobacterium or a mixture thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0061] FIG. 1 illustrates the results of an intraperitoneal glucose
tolerance test in adult male C57bl6 mice fed a high fat diet (HFD)
for four weeks or a normal chow (NC);
[0062] FIG. 2 illustrates the results of an intraperitoneal glucose
tolerance test four weeks after the beginning of the probiotic
treatment in high fat diet fed mice;
[0063] FIG. 3 illustrates the effect of treatment with
Bifidobacterium animalis subsp. lactis strain 420 (B420)
Lactobacillus acidophilus strain NCFM (NCFM) or a combination of
the two (B420+NCFM) on the body fat mass of high fat diet fed
mice;
[0064] FIG. 4 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the free water weight of high fat diet fed
mice;
[0065] FIG. 5 illustrates the effect of treatment with B420, NCFM
or 420+NCFM on the lean body mass weight of high fat diet fed
mice;
[0066] FIG. 6 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the total water weight of high fat diet fed
mice;
[0067] FIG. 7 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the subcutaneous adipose tissue weight of high fat
diet fed mice;
[0068] FIG. 8 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the mesenteric adipose tissue weight of high fat
diet fed mice;
[0069] FIG. 9 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the liver weight of high fat diet fed mice;
[0070] FIG. 10 illustrates the body weight gain before and after
treatment with B420, NCFM or B420+NCFM of high fat diet fed
mice;
[0071] FIG. 11 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the fasted and fed insulin levels of high fat diet
fed mice;
[0072] FIG. 12 illustrates the effect of treatment with B420, NCFM
or B420+NCFM on the insulin sensitivity of high fat diet fed
mice;
[0073] FIG. 13 illustrates the liver cytokine mRNA concentrations
in HFD diabetic mice treated with B420, NCFM or B420+NCFM and
control HFD-fed mice;
[0074] FIG. 14 illustrates the liver tissue inflammatory index in
HFD diabetic mice treated with B420, NCFM or B420+NCFM and control
HFD-fed mice;
[0075] FIG. 15 illustrates the skeletal muscle cytokine mRNA
concentrations in HFD diabetic mice treated with B420, NCFM or
B420+NCFM and control HFD-fed mice;
[0076] FIG. 16 illustrates the inflammatory index of skeletal
muscle tissues in HFD diabetic mice treated with B420, NCFM or
B420+NCFM and control HFD-fed mice.
[0077] FIG. 17 illustrates the subcutaneous adipose tissue cytokine
mRNA concentrations in HFD diabetic mice treated with B420, NCFM or
B420+NCFM and control HFD-fed mice;
[0078] FIG. 18 illustrates the inflammatory index of subcutaneous
adipose tissues in HFD diabetic mice treated with B420, NCFM or
B420+NCFM and control HFD-fed mice;
[0079] FIG. 19 illustrates the blood glucose levels of HFD-fed mice
treated with B420, a combination of B420 with polydextrose
(B420+PDX), metformin (MET) or a combination of polydextrose and
metformin (PDX+MET) and control HFD-treated mice;
[0080] FIG. 20 illustrates the plasma insulin levels of HFD-fed
mice treated with B420, B420+PDX, MET or a combination of B420 and
metformin (B420+MET) and control HFD-treated mice; and
[0081] FIG. 21 illustrates the HOMA-IR (homeostatic model of
insulin resistance) levels of HFD-fed mice treated with B420,
B420+PDX, MET, B420+MET, PDX+MET and control HFD-treated mice.
DETAILED DESCRIPTION OF THE INVENTION
Bacteria
[0082] The bacterium used in the present invention is selected from
a Bifidobacterium or a mixture thereof. Preferably the
Bifidobacterium to be used in the present invention is a
Bifidobacterium which is generally recognised as safe and, which is
preferably GRAS approved.
[0083] The bacterium may be used in any form capable of exerting
the effects described herein. For example, the bacteria may be
viable, dormant, inactivated or dead bacteria. Preferably, the
bacteria are viable bacteria.
[0084] The bacteria may comprise whole bacteria or may comprise
bacterial components. Examples of such components include bacterial
cell wall components such as peptidoglycan, bacterial nucleic acids
such as DNA and RNA, bacterial membrane components, and bacterial
structural components such as proteins, carbohydrates, lipids and
combinations of these such as lipoproteins, glycolipids and
glycoproteins.
[0085] The bacteria may also or alternatively comprise bacterial
metabolites. In this specification the term `bacterial metabolites`
includes all molecules produced or modified by the (probiotic)
bacteria as a result of bacterial metabolism during growth,
survival, persistence, transit or existence of bacteria during
probiotic product manufacture and storage and during
gastrointestinal transit in a mammal. Examples include all organic
acids, inorganic acids, bases, proteins and peptides, enzymes and
co-enzymes, amino acids and nucleic acids, carbohydrates, lipids,
glycoproteins, lipoproteins, glycolipids, vitamins, all bioactive
compounds, metabolites containing an inorganic component, and all
small molecules, for example nitrous molecules or molecules
containing a sulphurous acid.
[0086] Preferably the bacteria comprise whole bacteria, more
preferably whole viable bacteria.
[0087] Preferably, the Bifidobacterium used in accordance with the
present invention is one which is suitable for human and/or animal
consumption. A skilled person will be readily aware of specific
species and or strains of Bifidobacteria from within the genera
described herein which are used in the food and/or agricultural
industries and which are generally considered suitable for human
and/or animal consumption.
[0088] In the present invention, the Bifidobacterium used may be of
the same type (species and strain) or may comprise a mixture of
species and/or strains.
[0089] Suitable Bifidobacteria are selected from the species
Bifidobacterium lactis, Bifidobacterium bifidium, Bifidobacterium
longum, Bifidobacterium animalis, Bifidobacterium breve,
Bifidobacterium infantis, Bifidobacterium catenulatum,
Bifidobacterium pseudocatenulatum, Bifidobacterium adolescentis,
and Bifidobacterium angulatum, and combinations of any thereof.
[0090] Preferably, the Bifidobacterium used in the present
invention is of the species Bifidobacterium animalis. More
preferably, the Bifidobacterium used in the present invention is of
the species Bifidobacterium animalis subsp. lactis.
[0091] In a particularly preferred embodiment, the bacteria used in
the present invention are Bifidobacterium animalis subsp. lactis
strain 420 (B420). This strain is commercially available from
Danisco A/S.
[0092] In one embodiment, the bacterium used in the present
invention is a probiotic bacterium. In this specification the term
`probiotic bacterium` is defined as covering any non-pathogenic
bacterium which, when administered live in adequate amounts, confer
a health benefit on the host. These probiotic strains generally
have the ability to survive the passage through the upper part of
the digestive tract. They are non-pathogenic, non-toxic and
exercise their beneficial effect on health on the one hand via
ecological interactions with the resident flora in the digestive
tract, and on the other hand via their ability to influence the
immune system in a positive manner via the "GALT" (gut-associated
lymphoid tissue). Depending on the definition of probiotics, these
bacteria, when given in a sufficient number, have the ability to
progress live through the intestine, however they do not cross the
intestinal barrier and their primary effects are therefore induced
in the lumen and/or the wall of the gastrointestinal tract. They
then form part of the resident flora during the administration
period. This colonization (or transient colonization) allows the
probiotic bacteria to exercise a beneficial effect, such as the
repression of potentially pathogenic micro-organisms present in the
flora and interactions with the immune system of the intestine.
[0093] In preferred embodiments, the bacterium used in the present
invention is a probiotic Bifidobacterium.
[0094] In some embodiments, the Bifidobacterium is used in the
present invention together with a bacterium of the genus
Lactobacillus. A combination of Bifidobacterium and Lactobacillus
bacteria according to the present invention exhibits a synergistic
effect in certain applications (i.e. an effect which is greater
than the additive effect of the bacteria when used separately). For
example, combinations which, in addition to having effect on the
mammal as single components, may have beneficial effect on the
other components of the combination, for example by producing
metabolites which are then in turn used as an energy source by
other components of the combination, or maintaining physiological
conditions which favour the other components.
[0095] Typically, the Lactobacillus bacteria are selected from the
species Lactobacillus acidophilus, Lactobacillus casei,
Lactobacillus kefiri, Lactobacillus bifidus, Lactobacillus brevis,
Lactobacillus helveticus, Lactobacillus paracasei, Lactobacillus
rhamnosus, Lactobacillus salivarius, Lactobacillus curvatus,
Lactobacillus bulgaricus, Lactobacillus sakei, Lactobacillus
reuteri, Lactobacillus fermentum, Lactobacillus farciminis,
Lactobacillus lactis, Lactobacillus delbreuckii, Lactobacillus
plantarum, Lactobacillus paraplantarum, Lactobacillus crispatus,
Lactobacillus gasseri, Lactobacillus johnsonii and Lactobacillus
jensenii, and combinations of any thereof.
[0096] In preferred embodiments, the Lactobacillus bacterium used
in the present invention is a probiotic Lactobacillus.
[0097] Preferably, the Lactobacillus bacterium used in the present
invention of the species Lactobacillus acidophilus.
[0098] In a preferred embodiment, the Bifidobacterium is used in
the present invention together with a bacterium of the species
Lactobacillus acidophilus strain NCFM. Lactobacillus acidophilus
NCFM was deposited by Rhodia Chimie, France, at the American Type
Culture Collection as PTA-4797 on 15 Nov. 2002.
[0099] In a particularly preferred embodiment, the bacteria used in
the present invention comprise a combination of Bifidobacterium
animalis subsp. lactis strain 420 (B420) and Lactobacillus
acidophilus strain NCFM (PTA-4797).
Dosage
[0100] The Bifidobacterium (such as a strain of Bifidobacterium
animalis subsp. lactis, for example Bifidobacterium animalis subsp.
lactis strain 420 (B420), and (if present) the Lactobacillus (such
as a strain of Lactobacillus acidophilus, for example Lactobacillus
acidophilus strain NCFM) used in accordance with the present
invention may comprise from 10.sup.6 to 10.sup.12 CFU of bacteria/g
of support, and more particularly from 10.sup.8 to 10.sup.12 CFU of
bacteria/g of support, preferably 10.sup.9 to 10.sup.12 CFU/g for
the lyophilized form.
[0101] Suitably, the Bifidobacterium (such as a strain of
Bifidobacterium animalis subsp. lactis, for example Bifidobacterium
animalis subsp. lactis strain 420 (B420), and (if present) the
Lactobacillus (such as a strain of Lactobacillus acidophilus, for
example Lactobacillus acidophilus strain NCFM), may be administered
at a dosage of from about 10.sup.6 to about 10.sup.12 CFU of
microorganism/dose, preferably about 10.sup.8 to about 10.sup.12
CFU of microorganism/dose. By the term "per dose" it is meant that
this amount of microorganism is provided to a subject either per
day or per intake, preferably per day. For example, if the
microorganism is to be administered in a food product (for example
in a yoghurt)--then the yoghurt will preferably contain from about
10.sup.8 to 10.sup.12 CFU of the microorganism. Alternatively,
however, this amount of microorganism may be split into multiple
administrations each consisting of a smaller amount of microbial
loading--so long as the overall amount of microorganism received by
the subject in any specific time (for instance each 24 hour period)
is from about 10.sup.6 to about 10.sup.12 CFU of microorganism,
preferably 10.sup.6 to about 10.sup.12 CFU of microorganism.
[0102] In accordance with the present invention an effective amount
of at least one strain of a microorganism may be at least 10.sup.6
CFU of microorganism/dose, preferably from about 10.sup.6 to about
10.sup.12 CFU of microorganism/dose, preferably about 10.sup.8 to
about 10.sup.12 CFU of microorganism/dose.
[0103] In one embodiment, preferably the Bifidobacterium (such as a
strain of Bifidobacterium animalis subsp. lactis, for example
Bifidobacterium animalis subsp. lactisstrain 420 (B420), and (if
present) the Lactobacillus (such as a strain of Lactobacillus
acidophilus, for example Lactobacillus acidophilus strain NCFM),
may be administered at a dosage of from about 10.sup.6 to about
10.sup.12 CFU of microorganism/day, preferably about 10.sup.8 to
about 10.sup.12 CFU of microorganism/day. Hence, the effective
amount in this embodiment may be from about 10.sup.6 to about
10.sup.12 CFU of microorganism/day, preferably about 10.sup.8 to
about 10.sup.12 CFU of microorganism/day.
[0104] CFU stands for "colony-forming units". By `support` is meant
the food product, dietary supplement or the pharmaceutically
acceptable support.
[0105] When Bifidobacteria are used in the present invention
together with Lactobacilli, the bacteria may be present in any
ratio capable of achieving the desired effects of the invention
described herein. Typically, the Bifidobacteria to Lactobacilli
ratio (measured in terms of colony forming units) is in the range
1:100 to 100:1, suitably 1:50 to 50:1, preferably 1:20 to 20:1,
more preferably 1:10 to 10:1, still more preferably 1:5 to 5:1, yet
more preferably 1:3 to 3:1 and even more preferably 1:2 to 2:1 and
most preferably 1:1.5 to 1.5:1. In a particular example, the
Bifidobacteria to Lactobacilli ratio is 1:1.
[0106] In particular, when Bifidobacteria animalis subsp. lactis
strain 420 (B420) bacteria are used in the present invention
together with Lactobacillus acidophilus strain NCFM bacteria, the
bacteria may be present in any ratio capable of achieving the
desired effects of the invention described herein. Typically, the
ratio of Bifidobacteria animalis subsp. lactis strain 420 to
Lactobacillus acidophilus strain NCFM (measured in terms of colony
forming units) is in the range is in the range 1:100 to 100:1,
suitably 1:50 to 50:1, preferably 1:20 to 20:1, more preferably
1:10 to 10:1, still more preferably 1:5 to 5:1, yet more preferably
1:3 to 3:1 and even more preferably 1:2 to 2:1 and most preferably
1:1.5 to 1.5:1. In a particular example, the Bifidobacteria
animalis subsp. lactis strain 420 to Lactobacillus acidophilus
strain NCFM ratio is 1:1.
Subjects/Medical Indications
[0107] The Bifidobacteria (and, if present, the Lactobacilli) to
which the present invention relates are administered to a mammal,
including for example livestock (including cattle, horses, pigs,
chickens and sheep), and humans. In some aspects of the present
invention the mammal is a companion animal (including pets), such
as a dog or a cat for instance. In some aspects of the present
invention, the subject may suitably be a human.
[0108] The Bifidobacteria (and, if present, the Lactobacilli) to
which the present invention relates may be suitable for treating a
number of diseases or conditions in mammals (particularly humans).
In this specification the term "treatment" or "treating" refers to
any administration of the Bifidobacteria (and, if present,
Lactobacilli) according to the present invention and includes: (1)
preventing the specified disease from occurring in a mammal which
may be predisposed to the disease but does not yet experience or
display the pathology or symptomatology of the disease (including
prevention of one or more risk factors associated with the
disease); (2) inhibiting the disease in a mammal that is
experiencing or displaying the pathology or symptomatology of the
diseased (i.e., arresting further development of the pathology
and/or symptomatology), or (3) ameliorating the disease in a mammal
that is experiencing or displaying the pathology or symptomatology
of the diseased (i.e., reversing the pathology and/or
symptomatology).
[0109] The Bifidobacteria to which the present invention relates
are suitable for administration to both diabetic and obese mammals.
They could also be suitable for diabetic and non-obese mammals, as
well as to obese mammals possessing the risk factors for diabetes,
but not yet in a diabetic state. This aspect is discussed in more
detail below.
[0110] In particular, the use of Bifidobacteria according to the
present invention is suitable for the treatment of mammals
ingesting a high-fat diet. This aspect is discussed in more detail
below.
[0111] As described in more detail in the Examples below, the
Bifidobacteria used in the present invention have a number of
biological activities. In particular, the Bifidobacteria used in
the present invention are capable of normalising insulin
sensitivity, increasing fed insulin secretion, decreasing fasted
insulin secretion, improving glucose tolerance in a mammal. These
effects confer the potential for use in the treatment of diabetes
and diabetes-related conditions (in particular, Type 2 diabetes and
impaired glucose tolerance).
[0112] In particular, as described in more detail in the Examples
below, the Bifidobacteria used in combination with Lactobacillus
bacteria (particularly Lactobacillus acidophilus bacteria) in
accordance with the present invention have a number of biological
activities. In particular, the Bifidobacteria used in the present
invention are capable of increasing fed insulin secretion and
improving glucose tolerance in a mammal. These effects confer the
potential for use in the treatment of diabetes and diabetes-related
conditions (in particular, Type 2 diabetes and impaired glucose
tolerance).
[0113] In this specification the term `diabetes` includes all forms
of diabetes which, as noted above, is characterised by disordered
metabolism and abnormally high blood sugar (hyperglycaemia)
resulting from insufficient levels of the hormone insulin. The term
therefore includes Type 1 diabetes, Type 2 diabetes, gestational
diabetes, and impaired glucose tolerance. Type 1 diabetes is
characterised by loss of the insulin-producing beta cells of the
islets of Langerhans in the pancreas, leading to a deficiency of
insulin. Type 2 diabetes mellitus is characterised by insulin
resistance or reduced insulin sensitivity, combined with reduced
insulin secretion. Gestational diabetes is formally defined as "any
degree of glucose intolerance with onset or first recognition
during pregnancy". Impaired Glucose Tolerance (IGT) is a
pre-diabetic state of dysglycemia that is associated with insulin
resistance and increased risk of cardiovascular pathology.
According to the criteria of the World Health Organization and the
American Diabetes Association, impaired glucose tolerance is
defined as two-hour glucose levels of 140 to 199 mg per dL (7.8 to
11.0 mmol) on the 75-g oral glucose tolerance test. A patient is
said to be under the condition of IGT when he/she has an
intermediately raised glucose level after 2 hours, but less than
would qualify for type 2 diabetes mellitus. The fasting glucose may
be either normal or mildly elevated. IGT may precede type 2
diabetes mellitus by many years. IGT is also a risk factor for
mortality.
[0114] In addition, the Bifidobacteria used in the present
invention are capable of inducing weight loss and lowering body fat
mass (in particular, mesenteric fat mass). These effects confer the
potential for use in the treatment of obesity and controlling
weight gain and/or inducing weight loss in a mammal.
[0115] In particular, as described in more detail in the Examples
below, the Bifidobacteria used in combination with Lactobacillus
bacteria (particularly Lactobacillus acidophilus bacteria) in
accordance with the present invention are capable of inducing
weight loss and lowering body fat mass (in particular, mesenteric
fat mass). These effects confer the potential for use in the
treatment of obesity and controlling weight gain and/or inducing
weight loss in a mammal.
[0116] In this specification, the term obesity is linked to body
mass index (BMI). The body mass index (BMI) (calculated as weight
in kilograms divided by the square of height in metres) is the most
commonly accepted measurement for overweight and/or obesity. A BMI
exceeding 25 is considered overweight. Obesity is defined as a BM!
of 30 or more, with a BMI of 35 or more considered as serious
comorbidity obesity and a BMI of 40 or more considered morbid
obesity.
[0117] As noted above, the term "obesity" as used herein includes
obesity, comorbidity obesity and morbid obesity. Therefore, the
term "obese" as used here may be defined as a subject having a BMI
of more than or equal to 30. In some embodiments, suitably an obese
subject may have a BMI of more than or equal to 30, suitably 35,
suitably 40.
[0118] While the composition of the invention is particularly
suitable for use in patients who are both diabetic and obese, the
composition is also suitable for those who are diabetic but not
obese. It may also be suitable for use in obese patients possessing
the risk factors for diabetes, but not yet in a diabetic state, as
it could be expected that an obese person (but not diabetic), could
limit the metabolic consequences of his obesity, i.e. the diabetes
or at least insulino-resistance development.
[0119] In addition, the Bifidobacteria used in the present
invention may be used for treating metabolic syndrome in a mammal.
Metabolic syndrome is a combination of medical disorders that
increase the risk of developing cardiovascular disease and
diabetes. Metabolic syndrome is also known as metabolic syndrome X,
syndrome X, insulin resistance syndrome, Reaven's syndrome or CHAOS
(Australia).
[0120] There is currently no single accepted definition of
metabolic syndrome. The World Health Organization criteria (1999)
require presence of diabetes mellitus, impaired glucose tolerance,
impaired fasting glucose or insulin resistance, AND two of the
following:
[0121] blood pressure: .gtoreq.140/90 mmHg
[0122] dyslipidaemia: triglycerides (TG): .gtoreq.1.695 mmol/L and
high-density lipoprotein cholesterol (HDL-C).ltoreq.0.9 mmol/L
(male), .ltoreq.1.0 mmol/L (female)
[0123] central obesity: waist:hip ratio>0.90 (male); >0.85
(female), and/or body mass index>30 kg/m.sup.2
[0124] microalbuminuria: urinary albumin excretion ratio.gtoreq.20
mg/min or albumin: creatinine ratio.gtoreq.30 mg/g.
[0125] The European Group for the Study of Insulin Resistance
(1999) requires insulin resistance defined as the top 25% of the
fasting insulin values among non-diabetic individuals AND two or
more of the following:
[0126] central obesity: waist circumference.gtoreq.94 cm (male),
.gtoreq.80 cm (female)
[0127] dyslipidaemia: TG.gtoreq.2.0 mmol/L and/or HDL-C<1.0
mg/dL or treated for dyslipidaemia
[0128] hypertension: blood pressure.gtoreq.140/90 mmHg or
antihypertensive medication fasting plasma glucose 6.1 mmol/L
[0129] The US National Cholesterol Education Program (NCEP) Adult
Treatment Panel III (2001) requires at least three of the
following: central obesity: waist circumference 102 cm or 40 inches
(male), >88 cm or 36 inches (female)
[0130] dyslipidaemia: TG.gtoreq.1.695 mmol/L (150 mg/dl)
[0131] dyslipidaemia: HDL-C<40 mg/dL (male), <50 mg/dL
(female)
[0132] blood pressure.gtoreq.130/85 mmHg
[0133] fasting plasma glucose.gtoreq.6.1 mmol/L (110 mg/dl)
[0134] In further embodiments, the Bifidobacteria (and, if present,
the Lactobacilli) used in the present invention may be used to
lower tissue inflammation (particularly, although not exclusively,
liver tissue inflammation, muscle tissue inflammation and/or
adipose tissue inflammation) in a mammal.
[0135] In one embodiment, the Bifidobacteria (and, if present, the
Lactobacilli) used in the present invention may be used to lower
liver tissue inflammation. This confers the potential for the
application of the bacteria in the treatment of hepatitis, which is
characterised by the destruction of a number of liver cells and the
presence of inflammatory cells in the liver tissue.
[0136] Hepatitis can be divided into two subgroups according to its
duration: acute hepatitis (lasting less than six months) and
chronic hepatitis (lasting longer than six months). Hepatitis may
be also classified according to its cause: for example, hepatitis
may comprise Infectious viral hepatitis (such as hepatitis A,
hepatitis B, hepatitis C, hepatitis D and hepatitis E), hepatitis
caused by other viral diseases (such as mononucleosis and
cytomegalovirus), hepatitis caused by severe bacterial infections
or amoebic infections. hepatitis caused by medicines, hepatitis
caused by toxins such as alcohol, autoimmune hepatitis (in which a
number of liver cells are destroyed by the patient's own immune
system) and hepatitis caused by congenital metabolic disorders,
such as Wilson's disease (disorder of the body's copper metabolism)
and haemochromatosis (disorder of the body's iron metabolism).
[0137] In one embodiment, the Bifidobacteria (and, if present, the
Lactobacilli) used in the present invention may be used to lower
muscle tissue inflammation. This confers the potential for the
application of the bacteria in the treatment of myositis, in which
the muscle fibers and skin are inflamed and damaged, resulting in
muscle weakness.
[0138] There are several types of myositis that affect different
parts of the body. articular forms of myositis treatable according
to the present invention include: polymyositis (PM) (in which
muscles in many parts of the body, and especially those parts
closest to the trunk, are inflamed); dermatomyositis (DM) (which
affects both the muscle fibers and skin by damaging capillaries
that supply blood to the muscle and skin), inclusion body myositis
(IBM) which is characterized by gradual weakening of muscles
throughout the body, including the wrists or fingers, development
of dysphagia, and atrophy of forearms and/or thigh muscles; and
juvenile myositis (JM), which involves muscle weakness, skin rash,
and dysphagia in children.
[0139] The present inventors have surprisingly found that the
Bifidobacteria (and, if present, the Lactobacilli) to which the
present invention relates are capable of lowering adipose tissue
inflammation in mammals. There is epidemiological evidence in the
literature showing a statistical relationship between inflammation,
obesity and insulin resistance in humans (Cani et al., Diabetes,
2007, 56, 1761-1772, and references cited therein). This finding
therefore confers the potential for the Bifidobacteria (and, if
present, the Lactobacilli) to be useful in the treatment of
obesity, diabetes and related conditions, metabolic diseases and
cardiovascular consequences in mammals.
[0140] According to Berg and Scherer, Circulation Research, 2005,
96, 939, recent evidence highlights the role of adipose tissue in
the development of a systemic inflammatory state that contributes
to obesity-associated vasculopathy and cardiovascular risk.
[0141] Circulating mediators of inflammation participate in the
mechanisms of vascular insult and atheromatous change, and many of
these inflammatory proteins are secreted directly from adipocytes
and adipose tissue-derived macrophages. Several factors linking
obesity with an increased cardiovascular risk have been identified.
The adipocyte-specific secretory protein adiponectin is a
particularly promising candidate in this context. its levels are
decreased in obesity.
[0142] The targeted suppression of various proinflammatory cascades
in adipocytes specifically represents a new therapeutic opportunity
for the cardiovascular disease area. Suppression of adipose tissue
inflammation would therefore be expected to provide a therapeutic
benefit in the treatment of cardiovascular diseases.
[0143] Examples of cardiovascular diseases treatable by use of the
Bifidobacteria (and, if present, the Lactobacilli) according to the
present invention include aneurysm, angina, atherosclerosis,
cerebrovascular accident (stroke), cerebrovascular disease,
congestive heart failure (CHF), coronary artery disease, myocardial
infarction (heart attack) and peripheral vascular disease.
[0144] An aneurysm is a localized, blood-filled dilation
(balloon-like bulge) of a blood vessel caused by disease or
weakening of the vessel wall. Aneurysms most commonly occur in
arteries at the base of the brain (the circle of Willis) and in the
aorta (the main artery coming out of the heart, a so-called aortic
aneurysm). As the size of an aneurysm increases, there is an
increased risk of rupture, which can result in severe hemorrhage or
other complications including sudden death.
[0145] Angina pectoris, commonly known as angina, is severe chest
pain due to ischemia (a lack of blood and hence oxygen supply) of
the heart muscle, generally due to obstruction or spasm of the
coronary arteries (the heart's blood vessels). Coronary artery
disease, the main cause of angina, is due to atherosclerosis of the
cardiac arteries.
[0146] Atherosclerosis is the condition in which an artery wall
thickens as the result of a build up of fatty materials such as
cholesterol. It is a syndrome affecting arterial blood vessels. It
is a chronic inflammatory response in the walls of arteries, in
large part due to the accumulation of macrophage white blood cells
and promoted by low density (especially small particle)
lipoproteins (plasma proteins that carry cholesterol and
triglycerides) without adequate removal of fats and cholesterol
from the macrophages by functional high density lipoproteins (HDL).
It is commonly referred to as a hardening or furring of the
arteries. It is caused by the formation of multiple plaques within
the arteries.
[0147] A stroke is the rapidly developing loss of brain function(s)
due to disturbance in the blood supply to the brain. This can be
due to ischemia (lack of blood supply) caused by thrombosis or
embolism or due to a hemorrhage. As a result, the affected area of
the brain is unable to function, leading to inability to move one
or more limbs on one side of the body, inability to understand or
formulate speech, or see one side of the visual field and
ultimately to death.
[0148] Cerebrovascular disease is a group of brain dysfunctions
related to disease of blood vessels supplying the brain.
Hypertension is the most important cause that damages the blood
vessel lining endothelium exposing the underlying collagen where
platelets aggregate to initiate a repairing process which is not
always complete and perfect. Sustained hypertension permanently
changes the architecture of the blood vessels making them narrow,
stiff, deformed and uneven which are more vulnerable to
fluctuations of blood pressure. A fall in blood pressure during
sleep can lead to marked reduction in blood flow in the narrowed
blood vessels causing ischemic stroke in the morning whereas a
sudden rise in blood pressure can cause tearing of the blood
vessels causing intracranial hemorrhage during excitation at
daytime. Primarily people who are elderly, diabetic, smoker, or
have ischemic heart disease, have cerebrovascular disease. All
diseases related to artery dysfunction can be classified under a
disease as known as macrovascular disease. This is a simplistic
study by which arteries are blocked by fatty deposits or by a blood
clot. The results of cerebrovascular disease can include a stroke,
or even sometimes a hemorrhagic stroke. Ischemia or other blood
vessel dysfunctions can affect one during a cerebrovascular
accident.
[0149] Heart failure is a global term for the physiological state
in which cardiac output is insufficient for the body's needs. This
may occur when the cardiac output is low (often termed "congestive
heart failure"). Common causes of heart failure include myocardial
infarction and other forms of ischemic heart disease, hypertension,
valvular heart disease and cardiomyopathy.
[0150] Coronary disease (or coronary heart disease) refers to the
failure of coronary circulation to supply adequate circulation to
cardiac muscle and surrounding tissue. It is most commonly equated
with atherosclerotic coronary artery disease, but coronary disease
can be due to other causes, such as coronary vasospasm. It is
possible for the stenosis to be caused by the spasm.
[0151] Myocardial infarction, commonly known as a heart attack,
occurs when the blood supply to part of the heart is interrupted
causing some heart cells to die. This is most commonly due to
occlusion (blockage) of a coronary artery following the rupture of
a vulnerable atherosclerotic plaque, which is an unstable
collection of lipids (like cholesterol) and white blood cells
(especially macrophages) in the wall of an artery. The resulting
ischemia (restriction in blood supply) and oxygen shortage, if left
untreated for a sufficient period of time, can cause damage and/or
death (infarction) of heart muscle tissue (myocardium).
[0152] Peripheral vascular disease (PVD), also known as peripheral
artery disease (PAD) or peripheral artery occlusive disease (PAOD),
includes all diseases caused by the obstruction of large arteries
in the arms and legs. PVD can result from atherosclerosis,
inflammatory processes leading to stenosis, an embolism or thrombus
formation. It causes either acute or chronic ischemia (lack of
blood supply), typically of the legs.
[0153] It is envisaged within the scope of the present invention
that the embodiments of the invention can be combined such that
combinations of any of the features described herein are included
within the scope of the present invention. In particular, it is
envisaged within the scope of the present invention that any of the
therapeutic effects of the bacteria may be exhibited
concomitantly.
Diet
[0154] As noted above, diabetic and/or obese mammals treated with
bacteria according to the present invention may ingest a high-fat
diet while mitigating the metabolic consequences of their
condition(s). In this specification the term `high-fat diet` means
a diet generally containing at least 20%, preferably at least 25%,
such as at least 30%, for example at least 35%, such as at least
40%, for example at least 45%, such as at least 50%, for example at
least 55%, such as at least 60%, for example at least 65%, such as
at least 70%, for example at least 75%, such as at least 80%, for
example at least 85%, such as at least 90% of calories from
fat.
[0155] In some embodiments, mammals treated with bacteria according
to the present invention may ingest a low-carbohydrate diet during
the course of the treatment. In this specification the term
low-carbohydrate diet' means a diet generally containing no greater
than 50%, such as no greater than 45%, for example no greater than
40%, such as no greater than 35%, for example no greater than 30%,
such as no greater than 25%, for example no greater than 20%, such
as no greater than 15%, for example no greater than 10%, such as no
greater than 5%, for example no greater than 2%, such as no greater
than 1%, for example no greater than 0.5%, such as no greater than
0.2% of calories from carbohydrate.
Compositions
[0156] While is it possible to administer Bifidobacteria (and, if
present, Lactobacilli) alone according to the present invention
(i.e. without any support, diluent or excipient), the
Bifidobacteria (and, if present, Lactobacilli bacteria) are
typically and preferably administered on or in a support as part of
a product, in particular as a component of a food product, a
dietary supplement or a pharmaceutical formulation. These products
typically contain additional components well known to those skilled
in the art.
[0157] Any product which can benefit from the composition may be
used in the present invention. These include but are not limited to
foods, particularly fruit conserves and dairy foods and dairy
food-derived products, and pharmaceutical products. The
Bifidobacteria (and, if present, Lactobacilli) may be referred to
herein as "the composition of the present invention" or "the
composition".
Food
[0158] In one embodiment, the Bifidobacteria (and, if present,
Lactobacilli bacteria) are employed according to the invention in a
food product such as a food supplement, a drink or a powder based
on milk. Here, the term "food" is used in a broad sense and covers
food for humans as well s food for animals (i.e. a feed). In a
preferred aspect, the food is for human consumption.
[0159] The food may be in the form of a solution or as a
solid--depending on the use and/or the mode of application and/or
the mode of administration.
[0160] When used as, or in the preparation of, a food, such as
functional food, the composition of the present invention may be
used in conjunction with one or more of: a nutritionally acceptable
carrier, a nutritionally acceptable diluent, a nutritionally
acceptable excipient, a nutritionally acceptable adjuvant, a
nutritionally active ingredient.
[0161] By way of example, the composition of the present invention
can be used as an ingredient to soft drinks, a fruit juice or a
beverage comprising whey protein, health teas, cocoa drinks, milk
drinks and lactic acid bacteria drinks, yoghurt and drinking
yoghurt, cheese, ice cream, water ices and desserts, confectionery,
biscuits cakes and cake mixes, snack foods, balanced foods and
drinks, fruit fillings, care glaze, chocolate bakery filling,
cheese cake flavoured filling, fruit flavoured cake filling, cake
and doughnut icing, instant bakery filling creams, fillings for
cookies, ready-to-use bakery filling, reduced calorie filling,
adult nutritional beverage, acidified soy/juice beverage,
aseptic/retorted chocolate drink, bar mixes, beverage powders,
calcium fortified soy/plain and chocolate milk, calcium fortified
coffee beverage.
[0162] The composition can further be used as an ingredient in food
products such as American cheese sauce, anti-caking agent for
grated & shredded cheese, chip dip, cream cheese, dry blended
whip topping fat free sour cream, freeze/thaw dairy whipping cream,
freeze/thaw stable whipped tipping, low fat and light natural
cheddar cheese, low fat Swiss style yoghurt, aerated frozen
desserts, hard pack ice cream, label friendly, improved economics
& indulgence of hard pack ice cream, low fat ice cream: soft
serve, barbecue sauce, cheese dip sauce, cottage cheese dressing,
dry mix Alfredo sauce, mix cheese sauce, dry mix tomato sauce and
others.
[0163] The term "dairy product" as used herein is meant to include
a medium comprising milk of animal and/or vegetable origin. As milk
of animal origin there can be mentioned cow's, sheep's, goat's or
buffalo's milk. As milk of vegetable origin there can be mentioned
any fermentable substance of vegetable origin which can be used
according to the invention, in particular originating from
soybeans, rice or cereals.
[0164] Still more preferably the food product employed according to
the invention is a fermented milk or humanized milk.
[0165] For certain aspects, preferably the present invention may be
used in connection with yoghurt production, such as fermented
yoghurt drink, yoghurt, drinking yoghurt, cheese, fermented cream,
milk based desserts and others.
[0166] Suitably, the composition can be further used as an
ingredient in one or more of cheese applications, meat
applications, or applications comprising protective cultures.
[0167] The present invention also provides a method of preparing a
food or a food ingredient, the method comprising admixing the
composition according to the present invention with another food
ingredient.
[0168] Advantageously, the present invention relates to products
that have been contacted with the composition of the present
invention (and optionally with other components/ingredients),
wherein the composition is used in an amount to be capable of
improving the nutrition and/or health benefits of the product.
[0169] As used herein the term "contacted" refers to the indirect
or direct application of the composition of the present invention
to the product. Examples of the application methods which may be
used, include, but are not limited to, treating the product in a
material comprising the composition, direct application by mixing
the composition with the product, spraying the composition onto the
product surface or dipping the product into a preparation of the
composition.
[0170] Where the product of the invention is a foodstuff, the
composition of the present invention is preferably admixed with the
product. Alternatively, the composition may be included in the
emulsion or raw ingredients of a foodstuff. In a further
alternative, the composition may be applied as a seasoning, glaze,
colorant mixture, and the like.
[0171] For some applications, it is important that the composition
is made available on or to the surface of a product to be
affected/treated. This allows the composition to impart one or more
of the following favourable characteristics: nutrition and/or
health benefits.
[0172] The compositions of the present invention may be applied to
intersperse, coat and/or impregnate a product with a controlled
amount of a microorganism.
[0173] Preferably, the composition is used to ferment milk or
sucrose fortified milk or lactic media with sucrose and/or maltose
where the resulting media containing all components of the
composition--i.e. said microorganism according to the present
invention--can be added as an ingredient to yoghurt milk in
suitable concentrations--such as for example in concentrations in
the final product which offer a daily dose of 10.sup.6-10.sup.10
cfu. The microorganism according to the present invention may be
used before or after fermentation of the yoghurt.
[0174] For some aspects the microorganisms according to the present
invention are used as, or in the preparation of, animal feeds, such
as livestock feeds, in particular poultry (such as chicken) feed,
or pet food.
[0175] Advantageously, where the product is a food product, the
Bifidobacteria (and, if present, Lactobacilli) should remain
effective through the normal "sell-by" or "expiration" date during
which the food product is offered for sale by the retailer.
Preferably, the effective time should extend past such dates until
the end of the normal freshness period when food spoilage becomes
apparent. The desired lengths of time and normal shelf life will
vary from foodstuff to foodstuff and those of ordinary skill in the
art will recognise that shelf-life times will vary upon the type of
foodstuff, the size of the foodstuff, storage temperatures,
processing conditions, packaging material and packaging
equipment.
Food Ingredient
[0176] The composition of the present invention may be used as a
food ingredient and/or feed ingredient.
[0177] As used herein the term "food ingredient" or "feed
ingredient" includes a formulation which is or can be added to
functional foods or foodstuffs as a nutritional supplement.
[0178] The food ingredient may be in the form of a solution or as a
solid--depending on the use and/or the mode of application and/or
the mode of administration.
Food Supplements
[0179] The composition of the present invention may be--or may be
added to--food supplements (also referred to herein as dietary
supplements).
Functional Foods
[0180] The composition of the present invention may be--or may be
added to--functional foods.
[0181] As used herein, the term "functional food" means food which
is capable of providing not only a nutritional effect, but is also
capable of delivering a further beneficial effect to consumer.
[0182] Accordingly, functional foods are ordinary foods that have
components or ingredients (such as those described herein)
incorporated into them that impart to the food a specific
functional--e.g. medical or physiological benefit--other than a
purely nutritional effect.
[0183] Although there is no legal definition of a functional food,
most of the parties with an interest in this area agree that they
are foods marketed as having specific health effects beyond basic
nutritional effects.
[0184] Some functional foods are nutraceuticals. Here, the term
"nutraceutical" means a food which is capable of providing not only
a nutritional effect and/or a taste satisfaction, but is also
capable of delivering a therapeutic (or other beneficial) effect to
the consumer. Nutraceuticals cross the traditional dividing lines
between foods and medicine.
Medicament
[0185] The term "medicament" as used herein encompasses medicaments
for both human and animal usage in human and veterinary medicine.
In addition, the term "medicament" as used herein means any
substance which provides a therapeutic and/or beneficial effect.
The term "medicament" as used herein is not necessarily limited to
substances which need Marketing Approval, but may include
substances which can be used in cosmetics, nutraceuticals, food
(including feeds and beverages for example), probiotic cultures,
and natural remedies. In addition, the term "medicament" as used
herein encompasses a product designed for incorporation in animal
feed, for example livestock feed and/or pet food.
Pharmaceutical
[0186] The composition of the present invention may be used as--or
in the preparation of--a pharmaceutical. Here, the term
"pharmaceutical" is used in a broad sense--and covers
pharmaceuticals for humans as well as pharmaceuticals for animals
(i.e. veterinary applications). In a preferred aspect, the
pharmaceutical is for human use and/or for animal husbandry.
[0187] The pharmaceutical can be for therapeutic purposes--which
may be curative or palliative or preventative in nature. The
pharmaceutical may even be for diagnostic purposes.
[0188] A pharmaceutically acceptable support may be for example a
support in the form of compressed tablets, tablets, capsules,
ointments, suppositories or drinkable solutions. Other suitable
forms are provided below.
[0189] When used as--or in the preparation of--a pharmaceutical,
the composition of the present invention may be used in conjunction
with one or more of: a pharmaceutically acceptable carrier, a
pharmaceutically acceptable diluent, a pharmaceutically acceptable
excipient, a pharmaceutically acceptable adjuvant, a
pharmaceutically active ingredient.
[0190] The pharmaceutical may be in the form of a solution or as a
solid--depending on the use and/or the mode of application and/or
the mode of administration.
[0191] The Bifidobacteria (and, if present, Lactobacilli) of the
present invention may be used as pharmaceutical ingredients. Here,
the composition may be the sole active component or it may be at
least one of a number (i.e. 2 or more) of active components.
[0192] The pharmaceutical ingredient may be in the form of a
solution or as a solid--depending on the use and/or the mode of
application and/or the mode of administration.
[0193] The Bifidobacteria (and, if present, Lactobacilli) may be
used according to the present invention in any suitable
form--whether when alone or when present in a combination with
other components or ingredients. The lactic acid bacteria used in
the present invention may be referred to herein as "the
composition". Likewise, combinations comprising the composition of
the present invention and other components and/or ingredients (i.e.
ingredients--such as food ingredients, functional food ingredients
or pharmaceutical ingredients) may be used in any suitable
form.
[0194] The Bifidobacteria (and, if present, Lactobacilli) may be
used according to the present invention in the form of solid or
liquid preparations or alternatives thereof. Examples of solid
preparations include, but are not limited to tablets, capsules,
dusts, granules and powders which may be wettable, spray-dried or
freeze-dried. Examples of liquid preparations include, but are not
limited to, aqueous, organic or aqueous-organic solutions,
suspensions and emulsions.
[0195] Suitable examples of forms include one or more of: tablets,
pills, capsules, ovules, solutions or suspensions, which may
contain flavouring or colouring agents, for immediate-, delayed-,
modified-, sustained-, pulsed- or controlled-release
applications.
[0196] By way of example, if the composition of the present
invention is used in a tablet form--such for use as a functional
ingredient--the tablets may also contain one or more of: excipients
such as microcrystalline cellulose, lactose, sodium citrate,
calcium carbonate, dibasic calcium phosphate and glycine;
disintegrants such as starch (preferably corn, potato or tapioca
starch), sodium starch glycollate, croscarmellose sodium and
certain complex silicates; granulation binders such as
polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC),
hydroxypropylcellulose (HPC), sucrose, gelatin and acacia;
lubricating agents such as magnesium stearate, stearic acid,
glyceryl behenate and talc may be included.
[0197] Examples of nutritionally acceptable carriers for use in
preparing the forms include, for example, water, salt solutions,
alcohol, silicone, waxes, petroleum jelly, vegetable oils,
polyethylene glycols, propylene glycol, liposomes, sugars, gelatin,
lactose, amylose, magnesium stearate, talc, surfactants, silicic
acid, viscous paraffin, perfume oil, fatty acid monoglycerides and
diglycerides, petroethral fatty acid esters,
hydroxymethylcellulose, polyvinylpyrrolidone, and the like.
[0198] Preferred excipients for the forms include lactose, starch,
a cellulose, milk sugar or high molecular weight polyethylene
glycols.
[0199] For aqueous suspensions and/or elixirs, the composition of
the present invention may be combined with various sweetening or
flavouring agents, colouring matter or dyes, with emulsifying
and/or suspending agents and with diluents such as water, propylene
glycol and glycerin, and combinations thereof.
[0200] The forms may also include gelatin capsules; fibre capsules,
fibre tablets etc.; or even fibre beverages.
[0201] Further examples of form include creams. For some aspects
the microorganism used in the present invention may be used in
pharmaceutical and/or cosmetic creams such as sun creams and/or
after-sun creams for example.
[0202] In one aspect, the composition according to the present
invention may be administered in an aerosol, for example by way of
a nasal spray, for instance for administration to the respiratory
tract.
Combinations
[0203] The composition of the present invention may additionally
contain one or more prebiotics. Prebiotics are a category of
functional food, defined as non-digestible food ingredients that
beneficially affect the host by selectively stimulating the growth
and/or activity of one or a limited number of bacteria
(particularly, although not exclusively, probiotics, Bifidobacteria
and/or lactic acid bacteria) in the colon, and thus improve host
health. Typically, prebiotics are carbohydrates (such as
oligosaccharides), but the definition does not preclude
non-carbohydrates. The most prevalent forms of prebiotics are
nutritionally classed as soluble fibre. To some extent, many forms
of dietary fibre exhibit some level of prebiotic effect.
[0204] In one embodiment, a prebiotic is a selectively fermented
ingredient that allows specific changes, both in the composition
and/or activity in the gastrointestinal microflora that confers
benefits upon host well-being and health.
[0205] Suitably, the prebiotic may be used according to the present
invention in an amount of 0.01 to 100 g/day, preferably 0.1 to 50
g/day, more preferably 0.5 to 20 g/day. In one embodiment, the
prebiotic may be used according to the present invention in an
amount of 1 to 100 g/day, preferably 2 to 9 g/day, more preferably
3 to 8 g/day. In another embodiment, the prebiotic may be used
according to the present invention in an amount of 5 to 50 g/day,
preferably 10 to 25 g/day.
[0206] Examples of dietary sources of prebiotics include soybeans,
inulin sources (such as Jerusalem artichoke, jicama, and chicory
root), raw oats, unrefined wheat, unrefined barley and yacon.
[0207] Examples of suitable prebiotics include alginate, xanthan,
pectin, locust bean gum (LBG), inulin, guar gum,
galacto-oligosaccharide (GOS), fructo-oligosaccharide (FOS),
polydextrose (i.e. Litesse.RTM.), lactitol, lactosucrose, soybean
oligosaccharides, isomaltulose (Palatinose.TM.),
isomalto-oligosaccharides, gluco-oligosaccharides,
xylo-oligosaccharides, manno-oligosaccharides, beta-glucans,
cellobiose, raffinose, gentiobiose, melibiose, xylobiose,
cyclodextrins, isomaltose, trehalose, stachyose, panose, pullulan,
verbascose, galactomannans, and all forms of resistant starches. A
particularly preferred example of a prebiotic is polydextrose.
[0208] In some embodiments, a combination of Bifidobacterium (and,
if present, Lactobacillus) bacteria and prebiotics according to the
present invention exhibits a synergistic effect in certain
applications (i.e. an effect which is greater than the additive
effect of the bacteria when used separately). Without wishing to be
bound by theory, it is believed that such a combination is capable
of selectively stimulating the growth and/or activity of the
Bifidobacteria (and, if present, Lactobacilli) bacteria in the
colon, and thus improve host health.
[0209] In one embodiment, the Bifidobacteria (and, if present,
Lactobacilli) may be used according to the present invention in
combination with one or more antidiabetic drugs. Examples of oral
antidiabetic drugs which may be used in such a combination include
biguanides (such as metformin), sulfonylureas (such as carbutamide,
chlorpropamide, glibenclamide (Glyburide.TM.), gliclazide,
glimepiride, glipizide, gliquidone, tolazamide or tolbutamide),
alpha-glucosidase inhibitors (such as acarbose, miglitol or
voglibose), thiazolidinediones (TZD) (such as pioglitazone,
rivoglitazone or rosiglitazone), meglitinides (such as nateglinide,
repaglinide or mitiglinide), dipeptidyl peptidase-4 (DPP-4)
inhibitors (such as alogliptin, saxagliptin, sitagliptin or
vildagliptin), glucagon-like peptide-1 analogs (such as exenatide,
liraglutide, or albiglutide), amylin analogs (such as pramlintide),
fast acting insulin analogs (such as insulin lispro, insulin aspart
and insulin glulisine), long acting insulin analogs (such as
insulin glargine, insulin detemir), dual PPAR agonists (such as
aleglitazar) and SGLT2 inhibitors (such as dapagliflozin,
remogliflozin and sergliflozin). A particularly preferred example
is metformin.
[0210] The dosage, mode of administration and formulation of the
above antidiabetic drugs for use in the combination of the present
invention will be readily apparent to a skilled person. Suitably,
the antidiabetic drug may be used according to the present
invention in an amount of 1 .mu.g to 10 g/day, preferably 10 .mu.g
to 5 g/day, more preferably 0.1 mg to 2 g/day. In one embodiment,
the antidiabetic drug may be used according to the present
invention in an amount of 1 mg to 1 g/day, preferably 5 to 500
mg/day.
[0211] In one embodiment, the Bifidobacteria (and, if present,
Lactobacilli) may be used according to the present invention in
combination with both a prebiotic (as described and exemplified
above) and an antidiabetic drug (as described and exemplified
above).
[0212] Preferably, the Bifidobacterium used in the combination
(with a prebiotic, an antidiabetic drug, or both) is of the species
Bifidobacterium animalis. More preferably, the Bifidobacterium used
in the combination is of the species Bifidobacterium animalis
subsp. lactis. In a particularly preferred embodiment, the bacteria
used in the combination are Bifidobacterium animalis subsp. lactis
strain 420 (B420).
[0213] Suitably, the prebiotic used in the combination is
polydextrose.
[0214] Suitably, the antidiabetic used in the combination is
metformin.
[0215] In a particularly preferred embodiment, the bacteria used in
the combination are Bifidobacterium animalis subsp. lactis strain
420 (B420), the prebiotic is polydextrose and the antidiabetic is
metformin.
[0216] In another embodiment, a prebiotic may be used according to
the present invention in combination with an antidiabetic drug, but
in the absence of Bifidobacteria, Lactobacilli or other bacteria
used in the other embodiments of this invention.
[0217] Therefore, in a further aspect, the invention comprises a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof.
[0218] In a yet further aspect, the invention comprises a food
product or food product intermediate including a prebiotic or a
mixture thereof and an antidiabetic drug or a mixture thereof.
[0219] In a yet further aspect, the invention comprises a
pharmaceutical composition comprising a prebiotic or a mixture
thereof and an antidiabetic drug or a mixture thereof, together
with a pharmaceutically acceptable carrier or diluent.
[0220] In one aspect, the invention comprises use of a combination
of a prebiotic or a mixture thereof and an antidiabetic drug or a
mixture thereof in the manufacture of a food product, dietary
supplement or medicament for treating diabetes (preferably but not
exclusively Type 2 diabetes) in a mammal.
[0221] In another aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for treating impaired glucose
tolerance in a mammal.
[0222] In a further aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for normalising insulin
sensitivity in a mammal.
[0223] In a yet further aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for increasing fed insulin
secretion in a mammal.
[0224] In a still further aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for decreasing fasted insulin
secretion in a mammal.
[0225] In an additional aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for improving glucose tolerance in
a mammal.
[0226] In a yet further aspect, the invention comprises use of a
combination of a prebiotic or a mixture thereof and an antidiabetic
drug or a mixture thereof in the manufacture of a food product,
dietary supplement or medicament for treating metabolic syndrome in
a mammal.
[0227] In this embodiment, examples of suitable prebiotics which
may be used in such a combination include alginate, xanthan,
pectin, locust bean gum (LBG), inulin, guar gum,
galacto-oligosaccharide (GOS), fructo-oligosaccharide (FOS),
polydextrose (i.e. Litesse.RTM.), lactitol, lactosucrose, soybean
oligosaccharides, isomaltulose (Palatinose.TM.),
isomalto-oligosaccharides, gluco-oligosaccharides,
xylo-oligosaccharides, manno-oligosaccharides, beta-glucans,
cellobiose, raffinose, gentiobiose, melibiose, xylobiose,
cyclodextrins, isomaltose, trehalose, stachyose, panose, pullulan,
verbascose, galactomannans, and all forms of resistant starches. A
particularly preferred example of a prebiotic is polydextrose.
[0228] In this embodiment, examples of oral antidiabetic drugs
which may be used in such a combination include biguanides (such as
metformin), sulfonylureas (such as carbutamide, chlorpropamide,
glibenclamide (Glyburide.TM.), gliclazide, glimepiride, glipizide,
gliquidone, tolazamide or tolbutamide), alpha-glucosidase
inhibitors (such as acarbose, miglitol or voglibose),
thiazolidinediones (TZD) (such as pioglitazone, rivoglitazone or
rosiglitazone), meglitinides (such as nateglinide, repaglinide or
mitiglinide), dipeptidyl peptidase-4 (DPP-4) inhibitors (such as
alogliptin, saxagliptin, sitagliptin or vildagliptin),
glucagon-like peptide-1 analogs (such as exenatide, liraglutide, or
albiglutide), amylin analogs (such as pramlintide), fast acting
insulin analogs (such as insulin lispro, insulin aspart and insulin
glulisine), long acting insulin analogs (such as insulin glargine,
insulin detemir), dual PPAR agonists (such as aleglitazar) and
SGLT2 inhibitors (such as dapagliflozin, remogliflozin and
sergliflozin). A particularly preferred example is metformin. The
dosage, mode of administration and formulation of the above
antidiabetic drugs for use in the combination of this embodiment
will be readily apparent to a skilled person.
[0229] In this embodiment, the prebiotic used in the combination is
preferably polydextrose and the antidiabetic used in the
combination is metformin
[0230] In this combination, suitably, the prebiotic may be used in
an amount of 0.01 to 100 g/day, preferably 0.1 to 50 g/day, more
preferably 0.5 to 20 g/day. In one embodiment, the prebiotic may be
used in an amount of 1 to 100 g/day, preferably 2 to 9 g/day, more
preferably 3 to 8 g/day. In another embodiment, the prebiotic may
be used in an amount of 5 to 50 g/day, preferably 10 to 25
g/day.
[0231] In this combination, suitably, the antidiabetic drug may be
used in an amount of 1 to 10 g/day, preferably 10 .mu.g to 5 g/day,
more preferably 0.1 mg to 2 g/day. In one embodiment, the
antidiabetic drug may be used according to the present invention in
an amount of 1 mg to 1 g/day, preferably 5 to 500 mg/day.
EXAMPLE 1
Materials and Methods
Animal Model and Probiotic Treatment
[0232] A cohort of fifty C57B1/6 10-wk-old male mice were fed a
Normal Chow (NC) (A03, SAFE, Augy, France), or a high-fat diet
(HFD) (comprising 72% fat (corn oil and lard), 28% protein and
<1% carbohydrates) (SAFE, Augy, France) for 4 weeks. This diet
has the peculiar advantage to induce diabetes before the onset of
obesity (see for example Cani et al. 2008 "Role of gut microflora
in the development of obesity and insulin resistance following
high-fat diet feeding". Pathol Biol (Paris); Cani et al, Diabetes
2008, 57, 1470-81; Knauf et al. Endocrinology 2008, 149, 4768-77;
Cani et al., Diabetologia 2007, 50, 2374-83; Cani et al; Diabetes
2007, 56, 1761-1772 and Turini et al. Swiss Med Wkly 2007, 137,
700-4).
[0233] The mice underwent an intraperitoneal glucose tolerance
test. The area under curve was calculated and the mice dispatched
homogeneously according to the different experimental groups or ten
mice per group (10 mice per group). The mice were fed four more
weeks with a normal chow (n=10) or a HFD (n=40). The HFD mice were
treated daily for 4 weeks as follows with, 1. Vehicle treated, 2.
Bifidobacterium animalis subsp. lactis strain 420 (B420)
(10.sup.9/bacteria per mouse), 3. Lactobacillus acidophilus NCFM
(NCFM) (10.sup.9/bacteria per mouse), 4. NCFM+B420
(5.times.10.sup.8 B420+5.times.10.sup.8 NCFM per mouse). An
intraperitoneal test was then performed as described below. The
mice were housed in a controlled environment (inverted 12-h
daylight cycle, light off at 10:00 a.m.).
Weight Assessment
[0234] Mice were weighed weekly in the beginning of the study diet
administration (4 weeks prior to probiotic administration) until 6
weeks into the probiotic treatment (until sacrifice).
Body Composition
[0235] Mouse body composition was measured monthly using ECO-MRI.
Total body fat mass, total lean body mass, total water mass, free
water content, subcutaneous adipose tissue weight, mesenteric
adipose tissue weight and liver weight were measured.
Glucose Tolerance
[0236] Glucose tolerance was tested after 4-week administration the
study diets (before probiotic supplementation) to ensure the
glucose-intolerant and diabetic status of the HFD mice, and after 4
weeks of probiotic administration. Briefly, six-hour-fasted mice
were injected with 20% glucose (1 g/kg) into the peritoneal cavity.
Glycemia was determined with a glucose meter (ACCU-check Active,
Roche, Meylan, France) at 30 minutes before the glucose challenge,
at the time of the glucose challenge, and 30, 60 and 90 minutes
after the glucose challenge, from 3.5 .mu.L of tail-vein tip
collected blood.
Plasma Insulin
[0237] Insulin concentration was measured from plasma in fasted
state as well as in fed state.
Insulin Sensitivity
[0238] At completion of the probiotic treatment mice underwent an
intrafemoral surgery where a catheter was indwelled for further
intravenous infusions. This intravenous intrafemoral catheter was
implanted 4 days before the beginning of the experimental day
(infusions). The day of the assay the mice were fasted for 5 hours.
A hyperinsulinemic euglycemic clamp was performed for 3 hours in
the presence of tritiated labeled glucose to determine the glucose
turnover rate. Cold glucose was coinfused to maintain
euglycemia.
Inflammatory Markers (Real-Time Quantitative PCR)
[0239] The inflammation status of adipose, liver and muscle tissue
was measured by measuring the concentration of inflammatory markers
TNF.alpha., IL-1.beta., PAI-1, IL6 mRNAs by quantitative RT-PCR
analysis. Total mRNAs from the grafted fat pads and the recipient
subcutaneous adipose, liver and muscle tissue were extracted using
TriPure reagent (Roche, Basel, Switzerland). PCRs were performed
using an AbiPrism 7900 Sequence Detection System instrument and
software (Applied Biosystems, Foster City, Calif., USA, as
described in Cani et al. Diabetes 2007, 56, 1761-1772. The
concentration of each mRNA was normalized for RNA loading for each
sample using RPL19 rRNA as an internal standard.
Results
Glucose Tolerance
[0240] As shown in FIG. 1, all groups of mice fed a high fat diet
for 4 weeks were glucose intolerant and diabetic. Following B420
treatment, mice were characterized by an improved glucose tolerance
(FIG. 2). Significant decrease is achieved with B420 only; an trend
towards health benefit was obtained for NCFM alone or with a
combination of NCFM and B420. Therefore it was concluded that the
B420 treatment began to improve glucose tolerance. A longer period
of treatment could have had a greater impact on the glycemic
profiles.
Mouse Body Composition
[0241] The data show that four weeks of probiotic treatment with
B420 and a combination of B420 and NCFM reduced the impact of HFD
on body fat mass increase (FIG. 3). No effect of the treatment on
lean body mass, free water mass, and total water mass were observed
(FIGS. 4, 5 and 6).
[0242] In particular, FIG. 3 illustrates the effect of B420
treatment and combination of B420+NCFM in reducing the impact of
high fat diet on the increased body fat mass (wherein the left
column signifies the result before treatment and the right column
that after treatment). In FIG. 3, * indicates a result
significantly different from non treated mice of the same group. A
small increase of weight was observed with B420 or with the
combination of B420 and NCFM. However, this represents a
significant improvement on the untreated HFD mice, as the HFD
results show that ingestion of the HFD should increase
significantly the body fat mass of the mice.
[0243] As the treatment showed no effect on lean body mass, free
water mass, and total water mass, it can be concluded from the
above that the difference in weight achieved is only related to
adipose tissues.
Individual Tissue Weight
[0244] The data show that the mesenteric adipose tissue weight was
reduced by B420 and B420+NCFM. No differences in subcutaneous
adipose tissue or the liver weight were noted (FIGS. 7, 8 and
9).
[0245] In particular, FIG. 8 illustrates the effect on mesenteric
adipose tissue weight in high-fat diet fed mice treated with
probiotics. Significant were achieved for B420 alone or in
combination with NCFM (a P value of <0.05 was achieved when
compared with the untreated HFD group). The B420 and B420+NCFM
treated mice were characterized with less mesenteric fat mass.
[0246] In addition, FIG. 9 illustrates the effect on liver weight
in high fat diet fed mice treated with probiotics. No significant
difference was noted, once again showing that the weight difference
is not linked with the liver weight.
Weight Gain
[0247] FIG. 10 illustrates the body weight gain before and after
probiotic administration of high fat diet fed mice. Body weight
gain of the B420 and the B420+NCFM treated mice was lower than that
of the mice fed HFD without probiotics or mice treated with NCFM
alone. Statistically significant results were achieved for B420
alone or in combination with NCFM; a downward trend (in comparison
with untreated HFD-fed mice) was observed for NCFM alone.
Plasma Insulin Concentrations
[0248] Plasma insulin concentration was assessed in the fasted and
the fed state. The data show that, in fasting state, the B420
treated group was characterized by a normalization of fasting
hyperinsulinemia (FIG. 11). In the fed state all probiotic
treatments improved glucose insulin secretion.
[0249] These results are of significance, as low levels of insulin
(ie base levels of insulin) are observed in the fasted state of
healthy, non-diabetic subjects. Statistically significant results
were achieved for B420 alone.
[0250] Of further significance is that high levels of insulin are
observed in the fed state of healthy, non-diabetic subjects.
Statistically significant results were achieved for B420 alone,
NCFM alone and the combination of the two.
Insulin Sensitivity
[0251] FIG. 12 illustrates that high-fat diet fed mice were clamped
in hyperinsulinemic euglycemic condition by the clamp method. The
data show that B420 treated mice where characterized by a
normalization of insulin sensitivity.* p<0.05 vs HFD mice.
However, in the presence of NCFM this effect was not observed.
[0252] These results are of significance, as insulin sensitivity
provides the link between insulin behaviour and consumption of
glucose. The results shown with B420 are of particular interest
since, in comparison with classic anti-diabetic drugs which target
only fasted insulin, insulin sensitivity, or fed insulin, it has an
effect on all these factors.
Liver Tissue Inflammation
[0253] When considering all cytokine mRNA concentrations, HFD
induced inflammation in liver tissues (FIGS. 13 and 14). Probiotic
treatment had clear anti-inflammatory effect on the liver tissue.
This was particularly evident with NCFM treatment. Also treatment
with the combination of NCFM and B420 reduced in reduction of
inflammation, while B420 treatment alone reduced inflammation to
lesser extent.
Muscle Tissue Inflammation
[0254] Inflammation was induced by high fat diet also in muscle
tissues, although the induction of inflammation was not as strong
as in adipose tissue (FIGS. 15 and 16). Probiotic treatment with
B420+NCFM and NCFM alone tended to lower muscle tissue
inflammation, but the effect was not as clear as with adipose
tissue or liver tissue.
Adipose Tissue Inflammation
[0255] The high fat diet clearly induced inflammation in
subcutaneous adipose tissue (FIGS. 17 and 18). The B420 treatment
and the B420+NCFM treatment both showed strong anti-inflammatory
effect. Treatment with NCFM resulted in more inconsistent effects
on tissue inflammation but there was a general trend for reduced
inflammation.
[0256] Taken together, probiotic bacteria showed broad
anti-inflammatory effect, with most pronounced effects in adipose
tissue and liver tissue. It is notable that the anti-inflammatory
effects were differential and dependent on the tissue as well as
the probiotic treatment.
EXAMPLE 2
Materials and Methods
[0257] A cohort of C57BI/6 10-wk-old male mice were a high-fat diet
(HFD) (comprising 72% fat (corn oil and lard), 28% protein and
<1% carbohydrates) (SAFE, Augy, France) for 4 weeks as described
in Example 1. The mice underwent an intraperitoneal glucose
tolerance test. The area under curve was calculated and the mice
dispatched homogeneously according to the different experimental
groups or ten mice per group (10 mice per group). The mice were fed
four more weeks with HFD. The HFD mice were treated daily for 4
weeks as follows with B420 (10.sup.9 bacteria per mouse),
polydextrose (FDX) (0.2 g day), the antidiabetic drug metformin
(MET) (2 mg/mL drinking water), and various combinations of these.
Control mice were treated with saline. Mice were housed in a
controlled environment (inverted 12-h daylight cycle, light off at
10:00 a.m.). Blood glucose, insulin concentration and HOMA-IR were
measured from plasma in fasted state.
Results
[0258] Treatment either with B420 alone or the combination of B420
and polydextrose reduced fasting plasma glucose as compared to
control. Metformin alone did not have effect on fasting blood
glucose but a combination with metformin and polydextrose was
effective (FIG. 19).
[0259] Treatment with B420 reduced fasting plasma insulin. Addition
of polydextrose further improved the effect, suggesting a
synergistic effect of the combination. Metformin reduced the
fasting plasma insulin, but addition of B420 together with
metformin further improved the effect (FIG. 20).
[0260] Treatment with B420 reduced fasting HOMA-IR. Addition of
polydextrose with B420 further improved the effect, suggesting a
synergistic effect of the combination. Addition of metformin to
B420 or B420+polydextrose further improved the effect (FIG.
21).
[0261] All publications mentioned in the above specification are
herein incorporated by reference. Various modifications and
variations of the described methods and system of the present
invention will be apparent to those skilled in the art without
departing from the scope and spirit of the present invention.
Although the present invention has been described in connection
with specific preferred embodiments, it should be understood that
the invention as claimed should not be unduly limited to such
specific embodiments. Indeed, various modifications of the
described modes for carrying out the invention which are obvious to
those skilled in biochemistry and biotechnology or related fields
are intended to be within the scope of the following claims.
* * * * *