WO2001015715A2 - Composition for maintenance of bone or dental health or treatment of bone or dental disorders - Google Patents

Composition for maintenance of bone or dental health or treatment of bone or dental disorders Download PDF

Info

Publication number
WO2001015715A2
WO2001015715A2 PCT/EP2000/007919 EP0007919W WO0115715A2 WO 2001015715 A2 WO2001015715 A2 WO 2001015715A2 EP 0007919 W EP0007919 W EP 0007919W WO 0115715 A2 WO0115715 A2 WO 0115715A2
Authority
WO
WIPO (PCT)
Prior art keywords
lactobacillus
calcium
composition
bone
composition according
Prior art date
Application number
PCT/EP2000/007919
Other languages
French (fr)
Other versions
WO2001015715A3 (en
Inventor
Elisabeth Pasche
Fiona Ginty
Dominique Brassart
Jean-Richard Neeser
Original Assignee
Societe Des Produits Nestle S.A.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Societe Des Produits Nestle S.A. filed Critical Societe Des Produits Nestle S.A.
Priority to AU68385/00A priority Critical patent/AU6838500A/en
Publication of WO2001015715A2 publication Critical patent/WO2001015715A2/en
Publication of WO2001015715A3 publication Critical patent/WO2001015715A3/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/59—Compounds containing 9, 10- seco- cyclopenta[a]hydrophenanthrene ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease

Definitions

  • the present invention relates to a composition for maintenance of bone or dental health or treatment of bone or dental disorders, a method of preparing the composition; the composition for use as a medicament; use of the composition in the manufacture of a medicament or nutritional product; and a method of treatment of bone or dental disorders or maintenance of bone or dental health which comprises administering an effective amount of the composition.
  • FCA represents fractional calcium absorption
  • OC represents osteocalcin
  • PINP represents pro-collagen type I N-terminal propeptide
  • APPase represents alkaline phosphatase
  • DPYR represents deoxypyridinoline
  • sXlaps and uXlaps represent serum and urinary crosslaps, respectively.
  • FCA is determined after recovery of stable isotopes of calcium in urine samples.
  • OC, APPase, PINP and sXlaps are detected in blood samples.
  • DPYR and uXlaps are detected in urine samples.
  • FCA depicts intestinal calcium absorption.
  • OC, PINP and APPase are markers of bone formation.
  • DPYR, s- and uXlaps are markers of bone resorption.
  • Calcium is, for example, known to be of vital importance for the formation of bones and teeth, muscle contraction and the synthesis of hormones. Calcium is also an essential secondary messenger in several cellular activation pathways.
  • Minerals of which the diet is the primary source, are assimilated by the body by crossing the intestinal mucosa to pass into the blood stream.
  • the degree of assimilation (or of absorption) of minerals by the body depends both on their solubility in the intestinal medium and on the capacity of the intestinal cells to assimilate and transfer them into the blood stream.
  • Yaeshima et al (Bulletin of International Dairy Fermentation, No. 313, 1996) showed an increase in the absorption of calcium in rats from a diet of calcium fortified whey when a combination of oligosaccharides and Bifidobacteria was consumed.
  • Kot et al J, Agric. Food Chem., 43_, 1276-1282, 1995
  • Lactobacillus acidophilus naturally internalises Fe 2+ , and oxidises it to Fe 3+ which is insoluble and more difficult to assimilate.
  • osteoporosis is recognised as a major public health problem in Western countries, especially in post menopausal women.
  • post-menopausal osteoporotic fractures affect about 1.5 million people per annum.
  • About 300,000 new cases of osteoporotic hip, 650,000 vertebral and 200,000 distal forearm fractures are reported annually in the US.
  • Mortality in the first year after hip fractures reaches 20% (Amer. J. Epidem. 1001-1005 (1993)).
  • the estimated cost of treatment of these patients is in the region $6-$ 10 billion annually.
  • About half the survivors are unable to walk unassisted and 25% are confined to long term care in a nursing home. This problem is set to increase in view of the fact the life expectancy is increasing and a greater number of people are reaching old age.
  • composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium, calcium fortification compounds, calcium containing compounds and vitamin D has a positive affect on maintenance of bone or dental health or treatment of bone or dental disorders. This is unexpected because although the beneficial affect of calcium and/or vitamin D is well described it has now suprisingly been found that this affect may be increased in the presence of a probiotic bacterium.
  • the invention provides a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D.
  • the invention provides a method of producing a composition having a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D which comprises the steps of contacting a solution of calcium or a solution of vitamin D with a probiotic bacterium culture.
  • the invention provides a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D for use as a medicament.
  • the invention provides the use of a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D in the manufacture of a medicament or nutritive product for the treatment of a bone or dental disorder or maintenance of bone or dental health.
  • the invention provides a method of treatment of a bone or dental disorder or maintenance of bone or dental health which comprises administering an effective amount of a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D.
  • a composition according to an embodiment of the invention comprises a probiotic bacterium, calcium and vitamin D.
  • a composition according to an embodiment of the invention also comprises phosphorus and/or magnesium ions and/or calcium citrate malate (CCM).
  • CCM is well absorbed in man and provides the advantage of a high calcium intake or a higher bioavailability of calcium from the diet, even in diets deficient in calcium.
  • an embodiment of the composition according to the invention comprises casein phosphopeptide (CPP).
  • CPP provides the advantage of stimulating absorption of calcium intake from the diet, even where there is a deficiency of calcium in the diet.
  • the bacterium is a lactic acid bacterium. More preferably, it is selected from the group which comprises Lactobacillus acidophilus,
  • the bacterium is Lactobacillus acidophilus LalO (ATCC 11975).
  • An advantage provided by these bacteria is that they have a striking positive affect on enhancement of calcium absorption. Remarkably, the affects on bone formation and resorption are particularly striking when the composition comprises Lactobacillus acidophilus LalO (LalO).
  • a composition according to an embodiment of the invention comprises about 10 to about 10 cfu/ml or /g probiotic bacterium. More preferably it comprises about 10 cfu/ml probiotic bacterium.
  • a dose of about 250 to about 300g of a composition according to an embodiment of the invention is provided by two servings per day (eg. two yoghurts of about 125g to about 150g per day). More preferably the dose is about 250g per day ie two yoghurts of about 125g per day.
  • a composition according to an embodiment of the invention comprises about 15% to about 64% the RDA of calcium per 250g (RDA calcium is lOOOmg); ie about 75mg to about 320mg per 125g of the composition. More preferably it comprises about 50% of RDA calcium per 250g; ie 250mg per 125g of the composition.
  • a composition according to an embodiment of the invention comprises about l ⁇ g to about 3.75 ⁇ g vitamin D per 125g of the composition (yoghurt has been supplemented with 2.5 ⁇ g vitamin D/lOOg; thus, 2 such yoghurts of 125g provide more than 100% of RDA) . More preferably it comprises about 50% of RDA vitamin D per 250g (RDA vitamin D is 5 ⁇ g); ie 1.25 ⁇ g vitamin D per 125g of the composition.
  • an embodiment of a composition according to the invention comprises nutrients/non-nutrients selected from the group which comprises:
  • Vitamin D D3, A, Bl, B2, B6, B12, C, E, K, niacin, folic acid, pantothenic acid, biotin;
  • Prebiotics eg fibres, cellulose
  • Bioactive peptide derived from milk eg TGF- , PTHrp.
  • the amount of magnesium, calcium citrate malate or calcium citrate lactate or milk calcium, phosphorus, iron, iodine, zinc, copper, selenium is between about 15 and about 50% of RDA per 250g of the composition ie about 7.5 to about 25% RDA per 125g of the composition.
  • the amount of vitamin D, D3, A, Bl, B2, B6, B12, C, E, K, niacin, folic acid, pantothenic acid, biotin is between about 15 and about 33% of RDA per 125g of the composition.
  • the amount of prebiotics is about 6 to about 40g per 250g; ie about 3 to about 20g per 125g of the composition.
  • the amount of protein is about 2g to about 6g per 125g. More preferably it is about 4.4g per 125g of the composition.
  • the amount of casein phosphopeptide (CPP) is about 50 mg to about 2g per 250g of the composition; ie about 25mg to about lg per 125g of the composition.
  • the amount of isoflavones is about 25 to about 50 mg per 250g of the composition; ie about 12.5mg to about 25mg per 125g of the composition.
  • the amount of bioactive peptide derived from milk eg TGF- o r PTHrp is about 1.5 to about 2.5 ng per mg of protein.
  • Figure 1 shows results of a study on the positive effect of LalO on intestinal calcium absorption (FCA) and two bone formation markers (OC and PINP) in adult men.
  • Figure 2 shows results of a study on the effect of LalO on a bone formation marker (PINP) in post menopausal women.
  • Figure 3 shows the results of a study on the beneficial effect of a milk calcium preparation on bone mineral density in prepubertal girls. It is clear from the results shown in figure 1 that there is an improvement of calcium absorption and osteocalcin (bone formation) in the presence of lactic acid bacteria in men. In addition there is a trend for a decrease in deoxypyridinoline (DPYR) (bone resorption). In the light of the results obtained it may be concluded that a composition according to an embodiment of the invention may provide the benefit of maintenance of bone or dental health by positively affecting bone formation.
  • DTYR deoxypyridinoline
  • composition according to an embodiment of the invention may provide the benefit of maintenance of bone or dental health by positively affecting bone formation.
  • a composition according to an embodiment of the invention comprises about 250mg to about 320 mg calcium per 125g to 150g of the composition (which corresponds to about 30% calcium supplementation of a standard yoghurt; a standard yoghurt contains approx. 150 mg of calcium/ lOOg, i.e. 1.5 mg/g; a 30% supplementation would bring calcium concentration to 2 mg/g).
  • a composition in the form of a mixture of yoghurt, calcium derived from milk (LactovalTM (DMV International)), vitamin D and LalO lactic acid bacteria was prepared by mixture of the components. 0.16% LactovalTM, 0.001% vitamin D3 (Roche, 100CWS/A), and an appropriate amount of LalO bacteria were added to 125g yoghurts in order to reach 200 mg calcium, 3 ⁇ g vitamin D and 10 9 CFU LalO per lOOg of yoghurt. Tests were conducted to check for positive affects on calcium absorption and bone formation.
  • a composition was produced by simple mixture of yoghurt with LalO probiotic bacteria.
  • the minimal cell count of LalO was 10 7 /ml or g of yoghurt.
  • compositions which comprise the following were produced:
  • the main parameters analysed were intestinal calcium absorption using a stable isotope technique; Lactic Acid Bacteria survival using recovery of LalO and influence on endogenous flora; bone formation using osteocalcin, alkaline phosphatase and PINP analysis (osteocalcin, alkaline phosphatase and PINP were detected by immunoassays in blood samples); and bone resorption using serum and urinary crosslaps and DPYR analysis (serum and urinary crosslaps were detected in blood and urine samples, respectively; DPYR was detected by HPLC in urine samples.
  • Bone mineral density was determined by dual-energy X-ray absorptiometry at baseline, after 1 year of consumption, and after further 1 and 3.5 years without consumption of calcium- supplemented product.

Abstract

The application relates to a composition for maintenance of bone or dental health or treatment of bone or dental disorders, a method of preparing the composition, the composition for use as a medicament and a method of treatment. A preferred embodiment of the composition comprises La10 bacterium, vitamin D and calcium.

Description

Composition For Maintenance Of Bone Or Dental Health Or Treatment of Bone Or Dental Disorders
The present invention relates to a composition for maintenance of bone or dental health or treatment of bone or dental disorders, a method of preparing the composition; the composition for use as a medicament; use of the composition in the manufacture of a medicament or nutritional product; and a method of treatment of bone or dental disorders or maintenance of bone or dental health which comprises administering an effective amount of the composition.
Within the context of this specification the word "comprises" is taken to mean "includes, among other things". It is not intended to be construed as "consists of only".
Within the context of this specification RDA represents recommended daily amount, FCA represents fractional calcium absorption, OC represents osteocalcin, PINP represents pro-collagen type I N-terminal propeptide, APPase represents alkaline phosphatase, DPYR represents deoxypyridinoline, sXlaps and uXlaps represent serum and urinary crosslaps, respectively. FCA is determined after recovery of stable isotopes of calcium in urine samples. OC, APPase, PINP and sXlaps are detected in blood samples. DPYR and uXlaps are detected in urine samples. FCA depicts intestinal calcium absorption. OC, PINP and APPase are markers of bone formation. DPYR, s- and uXlaps are markers of bone resorption.
It is well known that minerals are key elements in several major physiological processes. Calcium is, for example, known to be of vital importance for the formation of bones and teeth, muscle contraction and the synthesis of hormones. Calcium is also an essential secondary messenger in several cellular activation pathways.
Minerals, of which the diet is the primary source, are assimilated by the body by crossing the intestinal mucosa to pass into the blood stream. The degree of assimilation (or of absorption) of minerals by the body depends both on their solubility in the intestinal medium and on the capacity of the intestinal cells to assimilate and transfer them into the blood stream.
The location, efficiency and the mechanisms of calcium absorption along the intestine have been studied in rats and chickens for many years. However, for ethical and technical reason such studies have been limited in man.
One of the most widely studied aspects of mineral absorption is the bioavailability of the minerals in relation to the composition of the daily diet. However, many minerals which are highly bioavailable also cause organoleptic degradation in some products and may be unsuitable for use in such products. Furthermore, merely adding greater amounts of minerals often has a negative effect on the organoleptic quality of the product.
Rasic et al (Fermented Fresh Milk Product, vol 1, pi 14-115, 1978) reported that minerals contained in dairy products are assimilated better when the products are fermented. This effect is attributed to the presence of acids in fermented dairy products.
More recently, Yaeshima et al (Bulletin of International Dairy Fermentation, No. 313, 1996) showed an increase in the absorption of calcium in rats from a diet of calcium fortified whey when a combination of oligosaccharides and Bifidobacteria was consumed. However, Kot et al (J, Agric. Food Chem., 43_, 1276-1282, 1995) reported that Lactobacillus acidophilus naturally internalises Fe2+, and oxidises it to Fe3+ which is insoluble and more difficult to assimilate.
In addition to the problems described above, osteoporosis is recognised as a major public health problem in Western countries, especially in post menopausal women. Indeed, post-menopausal osteoporotic fractures affect about 1.5 million people per annum. About 300,000 new cases of osteoporotic hip, 650,000 vertebral and 200,000 distal forearm fractures are reported annually in the US. Mortality in the first year after hip fractures reaches 20% (Amer. J. Epidem. 1001-1005 (1993)). The estimated cost of treatment of these patients is in the region $6-$ 10 billion annually. About half the survivors are unable to walk unassisted and 25% are confined to long term care in a nursing home. This problem is set to increase in view of the fact the life expectancy is increasing and a greater number of people are reaching old age.
Remarkably, it has now been found that a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium, calcium fortification compounds, calcium containing compounds and vitamin D has a positive affect on maintenance of bone or dental health or treatment of bone or dental disorders. This is unexpected because although the beneficial affect of calcium and/or vitamin D is well described it has now suprisingly been found that this affect may be increased in the presence of a probiotic bacterium.
Accordingly, in a first aspect the invention provides a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D.
In a second aspect, the invention provides a method of producing a composition having a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D which comprises the steps of contacting a solution of calcium or a solution of vitamin D with a probiotic bacterium culture.
In a third aspect the invention provides a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D for use as a medicament.
In a fourth aspect the invention provides the use of a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D in the manufacture of a medicament or nutritive product for the treatment of a bone or dental disorder or maintenance of bone or dental health.
In a fifth aspect the invention provides a method of treatment of a bone or dental disorder or maintenance of bone or dental health which comprises administering an effective amount of a composition which comprises a probiotic bacterium and at least one component selected from the group consisting of calcium and vitamin D. Preferably a composition according to an embodiment of the invention comprises a probiotic bacterium, calcium and vitamin D. More preferably a composition according to an embodiment of the invention also comprises phosphorus and/or magnesium ions and/or calcium citrate malate (CCM). CCM is well absorbed in man and provides the advantage of a high calcium intake or a higher bioavailability of calcium from the diet, even in diets deficient in calcium.
Preferably, an embodiment of the composition according to the invention comprises casein phosphopeptide (CPP). CPP provides the advantage of stimulating absorption of calcium intake from the diet, even where there is a deficiency of calcium in the diet.
Preferably, the bacterium is a lactic acid bacterium. More preferably, it is selected from the group which comprises Lactobacillus acidophilus,
Lactobacillus crispatus, Lactobacillus amylovorous, Lactobacillus gallinarum, Lactobacillus gas se and Lactobacillus johnsonii; Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus helveticus, Lactobacillus brevis; Lactobacillus fermentum; Lactobacillus plantarum; Lactobacillus casei particularly L. casei, Lactobacillus paracasei and L. rhamnosus; Lactobacillus delbrueckii particularly L. delbrueckii subsp lactis, L. delbrueckii subsp. delbrueckii and L. delbrueckii subsp. bulgaricus; and bifidobacteria particularly Bifidobacterium infantis, Bifidobacterium breve, Biβdobacterium adolescentis, Bifidobacterium lactis Biβdobacterium longum; and Leuconostoc mesenteroides particularly L. mesenteroides subsp cremoris. Most preferably the bacterium is Lactobacillus acidophilus LalO (ATCC 11975). An advantage provided by these bacteria is that they have a striking positive affect on enhancement of calcium absorption. Remarkably, the affects on bone formation and resorption are particularly striking when the composition comprises Lactobacillus acidophilus LalO (LalO).
Preferably a composition according to an embodiment of the invention comprises about 10 to about 10 cfu/ml or /g probiotic bacterium. More preferably it comprises about 10 cfu/ml probiotic bacterium. Preferably a dose of about 250 to about 300g of a composition according to an embodiment of the invention is provided by two servings per day (eg. two yoghurts of about 125g to about 150g per day). More preferably the dose is about 250g per day ie two yoghurts of about 125g per day.
Preferably a composition according to an embodiment of the invention comprises about 15% to about 64% the RDA of calcium per 250g (RDA calcium is lOOOmg); ie about 75mg to about 320mg per 125g of the composition. More preferably it comprises about 50% of RDA calcium per 250g; ie 250mg per 125g of the composition.
Preferably, a composition according to an embodiment of the invention comprises about l μg to about 3.75μg vitamin D per 125g of the composition (yoghurt has been supplemented with 2.5 μg vitamin D/lOOg; thus, 2 such yoghurts of 125g provide more than 100% of RDA) . More preferably it comprises about 50% of RDA vitamin D per 250g (RDA vitamin D is 5μg); ie 1.25μg vitamin D per 125g of the composition.
Preferably, an embodiment of a composition according to the invention comprises nutrients/non-nutrients selected from the group which comprises:
Magnesium, calcium citrate malate or calcium citrate lactate or milk calcium, phosphorus, iron, iodine, zinc, copper, selenium;
Vitamin D, D3, A, Bl, B2, B6, B12, C, E, K, niacin, folic acid, pantothenic acid, biotin;
Prebiotics (eg fibres, cellulose);
Protein, casein phosphopeptide (CPP);
Isoflavones;
Bioactive peptide derived from milk eg TGF- , PTHrp.
Preferably the amount of magnesium, calcium citrate malate or calcium citrate lactate or milk calcium, phosphorus, iron, iodine, zinc, copper, selenium is between about 15 and about 50% of RDA per 250g of the composition ie about 7.5 to about 25% RDA per 125g of the composition.
Preferably the amount of vitamin D, D3, A, Bl, B2, B6, B12, C, E, K, niacin, folic acid, pantothenic acid, biotin is between about 15 and about 33% of RDA per 125g of the composition.
Preferably the amount of prebiotics (eg fibres, cellulose)is about 6 to about 40g per 250g; ie about 3 to about 20g per 125g of the composition.
Preferably the amount of protein is about 2g to about 6g per 125g. More preferably it is about 4.4g per 125g of the composition.
Preferably the amount of casein phosphopeptide (CPP) is about 50 mg to about 2g per 250g of the composition; ie about 25mg to about lg per 125g of the composition.
Preferably the amount of isoflavones is about 25 to about 50 mg per 250g of the composition; ie about 12.5mg to about 25mg per 125g of the composition.
Preferably the amount of bioactive peptide derived from milk eg TGF- o r PTHrp is about 1.5 to about 2.5 ng per mg of protein.
Specific embodiments of the invention will now be described in further detail with reference to the accompanying drawings in which:
Figure 1 shows results of a study on the positive effect of LalO on intestinal calcium absorption (FCA) and two bone formation markers (OC and PINP) in adult men.
Figure 2 shows results of a study on the effect of LalO on a bone formation marker (PINP) in post menopausal women.
Figure 3 shows the results of a study on the beneficial effect of a milk calcium preparation on bone mineral density in prepubertal girls. It is clear from the results shown in figure 1 that there is an improvement of calcium absorption and osteocalcin (bone formation) in the presence of lactic acid bacteria in men. In addition there is a trend for a decrease in deoxypyridinoline (DPYR) (bone resorption). In the light of the results obtained it may be concluded that a composition according to an embodiment of the invention may provide the benefit of maintenance of bone or dental health by positively affecting bone formation.
It is clear from the results shown in figure 2 that there is an improvement of PINP (bone formation) in women in the presence of lactic acid bacteria.
In the light of the results obtained it may be concluded that a composition according to an embodiment of the invention may provide the benefit of maintenance of bone or dental health by positively affecting bone formation.
Example 1
A composition according to an embodiment of the invention comprises about 250mg to about 320 mg calcium per 125g to 150g of the composition (which corresponds to about 30% calcium supplementation of a standard yoghurt; a standard yoghurt contains approx. 150 mg of calcium/ lOOg, i.e. 1.5 mg/g; a 30% supplementation would bring calcium concentration to 2 mg/g).
A composition in the form of a mixture of yoghurt, calcium derived from milk (Lactoval™ (DMV International)), vitamin D and LalO lactic acid bacteria was prepared by mixture of the components. 0.16% Lactoval™, 0.001% vitamin D3 (Roche, 100CWS/A), and an appropriate amount of LalO bacteria were added to 125g yoghurts in order to reach 200 mg calcium, 3 μg vitamin D and 109 CFU LalO per lOOg of yoghurt. Tests were conducted to check for positive affects on calcium absorption and bone formation.
Enrichment with Ca and Vitamin D was carried out during standardisation of milk. Both Ca and Vitamin D were mixed with the milk before pasteurisation and homogenisation. Example 2
A composition was produced by simple mixture of yoghurt with LalO probiotic bacteria. The minimal cell count of LalO was 107/ml or g of yoghurt.
Example 3
Compositions which comprise the following were produced:
Milk calcium (Lactoval™ (DMV International)) in cakes;
Milk calcium (Lactoval™ (DMV International)) in biscuits;
Milk calcium (Lactoval™ (DMV International)) in chocolate bars;
Milk calcium (Lactoval™ (DMV International)) in fruit juices;
Milk calcium (Lactoval™ (DMV International)) in powdered drinking chocolate;
Milk calcium (Lactoval™ (DMV International)) in yoghurts;
In vitro data on LalO and Ca absorption and in vivo data on the effects of LalO on bone metabolism were obtained. It is presented in the figures.
The protocols for the in vivo studies were as follows:
In a first study, a two week wash-out period was followed by two weeks of test followed by three weeks of wash-out and two further weeks of test . 10 male and 21 female subjects were enrolled in a cross-over design so that each volunteer received yoghurt with LalO during one test period and yoghurt without LalO during the other test period. The study was designed to detect a 6% absolute increase in calcium absorption from the diet. One strain of LalO bacterium was used. The main parameters analysed were intestinal calcium absorption using a stable isotope technique; Lactic Acid Bacteria survival using recovery of LalO and influence on endogenous flora; bone formation using osteocalcin, alkaline phosphatase and PINP analysis (osteocalcin, alkaline phosphatase and PINP were detected by immunoassays in blood samples); and bone resorption using serum and urinary crosslaps and DPYR analysis (serum and urinary crosslaps were detected in blood and urine samples, respectively; DPYR was detected by HPLC in urine samples.
The study on men revealed the following:
With regard to calcium absorption, there was a trend for an increase.
With regard to LalO recovery and influence on endogenous flora, there was a 50% recovery of LalO and a significant increase in total lactobacilli population.
With regard to bone turnover, there was a significant increase in osteocalcin, a bone formation marker, and a trend for a decrease in DPYR, a bone resorption marker.
The study on women revealed the following:
With regard to calcium absorption, variability of the results was slightly lower than expected, mean absorption was lower than in men.
With regard to LalO recovery, similar results to the study in men were achieved.
With regard to bone turnover, there was a significant increase in PINP, a bone formation marker
In a second study, 149 pre-pubertal girls received calcium-supplemented products, listed in Example 3, for 1 year. Bone mineral density (BMD) was determined by dual-energy X-ray absorptiometry at baseline, after 1 year of consumption, and after further 1 and 3.5 years without consumption of calcium- supplemented product.
The study on pre-pubertal girls revealed the following:
Girls receiving the calcium-supplemented products had a significantly higher gain in BMD. Gains in BMD were maintained at least for 3.5 years.
In general the studies revealed the following results:
Milk Calcium
Helps increase bone mineral density during growth
LalO bacterium
Helps increase the amount of Ca absorbed Helps decrease bone loss Helps increase bone formation
LalO bacterium + milk Ca + vitamin D
Helps increase the amount of Ca absorbed Helps increase Ca absorption Helps decrease bone loss
Helps increase bone formation
Helps increase bone mineral density during growth
In the light of these results it was concluded that the results in men and in women show that bone formation may be significantly increased, and that the results in girls show that BMD may be significantly increased, if a diet including a composition according to the invention is administered.
It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present invention and without diminishing its attendant advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

Claims

Claims
1. A composition which comprises a probiotic bacterium, and at least one component selected from the group consisting of calcium and vitamin D.
2. A composition according to claim 1 which comprises calcium and vitamin D.
3. A composition according to claim 1 or 2 wherein the probiotic bacterium is selected from the group which comprises Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus amylovorous, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus johnsonii; Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus helveticus, Lactobacillus brevis; Lactobacillus fermentum; Lactobacillus plantarum; Lactobacillus casei particularly L. casei, Lactobacillus paracasei and L. rhamnosus; Lactobacillus delbrueckii, L. delbrueckii subsp lactis, L. delbrueckii subsp.
Delbrueckii, L. delbrueckii subsp. bulgaricus; bifidobacteria, Biβdobacterium infantis, Biβdobacterium breve, Biβdobacterium adolescentis, Biβdobacterium lactis Biβdobacterium longum; and Leuconostoc mesenteroides, L. mesenteroides subsp cremoris.
4. A composition according to any preceding claim which comprises about 10 to about 109 cfu/ml or /g probiotic bacterium.
5. A composition according to any preceding claim which comprises about 75mg to about 320mg calcium per 125g of the composition.
6. A composition according to any preceding claim which comprises about lμg to about 3.75μg vitamin D per 125g of the composition.
7. A method of producing a composition having a probiotic bacterium, calcium and vitamin D which comprises the steps of contacting a solution of calcium with a solution of vitamin D with a probiotic bacterium culture.
8. A composition according to any one of claims 1 to 6 for use as a medicament.
. Use of a composition according to any one of claims 1 to 6 in the manufacture of a medicament or nutritive product for the treatment of a bone disorder or maintenance of bone health.
10. A method of treatment of a bone disorder or maintenance of bone health which comprises administering an effective amount of a composition according to any one of claims 1 to 6.
PCT/EP2000/007919 1999-08-31 2000-08-14 Composition for maintenance of bone or dental health or treatment of bone or dental disorders WO2001015715A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU68385/00A AU6838500A (en) 1999-08-31 2000-08-14 Composition for maintenance of bone or dental health or treatment of bone or dental disorders

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB9920578.3A GB9920578D0 (en) 1999-08-31 1999-08-31 Composition for maintenance of bone or dental health or treatment of bone or dental disorders
GB9920578.3 1999-08-31

Publications (2)

Publication Number Publication Date
WO2001015715A2 true WO2001015715A2 (en) 2001-03-08
WO2001015715A3 WO2001015715A3 (en) 2001-10-18

Family

ID=10860095

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2000/007919 WO2001015715A2 (en) 1999-08-31 2000-08-14 Composition for maintenance of bone or dental health or treatment of bone or dental disorders

Country Status (3)

Country Link
AU (1) AU6838500A (en)
GB (1) GB9920578D0 (en)
WO (1) WO2001015715A2 (en)

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1230920A1 (en) * 1999-10-19 2002-08-14 Meiji Dairies Corporation Food materials useful in preventing and ameliorating metabolic bone diseases and preventives/remedies for metabolic bone diseases comprising these materials
JP2003300900A (en) * 2002-04-11 2003-10-21 Lotte Co Ltd Calcium absorption promoter, promoter of crystal growth of calcium phosphate, and food and drink and fodder containing them
JP2004315477A (en) * 2003-04-18 2004-11-11 Tokyoto Koreisha Kenkyu Fukushi Shinko Zaidan Bone resorption inhibitor
US6872565B2 (en) 2003-01-29 2005-03-29 Biogaia Ab Product containing Lactobacillus reuteri strain ATTC PTA-4965 or PTA-4964 for inhibiting bacteria causing dental caries
EP1609463A1 (en) * 2004-06-23 2005-12-28 L'oreal Process and compositions for the protection and/or the treatment of dry and/or sensitive skin
WO2006000992A1 (en) * 2004-06-23 2006-01-05 L'oreal Method and compositions useful for preventing and/or treating sensitive and/or dry skin
US7517681B2 (en) 2003-01-29 2009-04-14 Biogaia Ab Selection and use of lactic acid bacteria for reducing dental caries and bacteria causing dental caries
US7956031B2 (en) 2005-05-31 2011-06-07 Naidu Lp Metallo-lactoferrin-coenzyme compositions for trigger and release of bioenergy
WO2011066880A2 (en) 2009-09-17 2011-06-09 Danone, S.A. Pasteurised and fermented dairy product and procedure for preparing such product
US8021659B2 (en) * 2006-04-28 2011-09-20 Naidu Lp Coenzyme Q10, lactoferrin and angiogenin compositions and uses thereof
ES2365145A1 (en) * 2009-09-17 2011-09-23 Danone, S.A. Fermented pasterised dairy product and preparation procedure of the same. (Machine-translation by Google Translate, not legally binding)
US20110280840A1 (en) * 2010-05-04 2011-11-17 New York University Compositions and methods for treating obesity and related disorders by characterizing and restoring mammalian bacterial microbiota
US8481299B2 (en) 2009-03-04 2013-07-09 L'oreal Use of probiotic microorganisms to limit skin irritation
WO2014163568A1 (en) * 2013-04-03 2014-10-09 Probi Ab Probiotic strains for use in treatment or prevention of osteoporosis
CN104152354A (en) * 2013-05-14 2014-11-19 仙农生物科技(上海)有限公司 Compound probiotics as well as production process and application thereof to leavening agent
CN104186655A (en) * 2014-08-27 2014-12-10 内蒙古伊利实业集团股份有限公司 Low-fat fermented milk having blood fat reduction and anti-oxidization functions and preparation method thereof
US9386793B2 (en) 2010-08-20 2016-07-12 New York University Compositions and methods for treating obesity and related disorders by characterizing and restoring mammalian bacterial microbiota
US9603876B2 (en) 2008-09-25 2017-03-28 New York University Compositions and methods for restoring gastrointestinal microbiota following antibiotic treatment
CN106858605A (en) * 2016-12-26 2017-06-20 吉林舒润生物科技有限公司 A kind of probiotics calcium functional food for preventing and treating bone information relevant disease
US9687513B2 (en) 2005-07-05 2017-06-27 Probi Ab Methods of treating anemia and increasing the absorption of non-heme iron by administration of lactobacillus plantarum
CN108245479A (en) * 2017-09-19 2018-07-06 王亚东 A kind of facial mask containing bifidobacterium lactis fermentation activity extract
US10238897B2 (en) 2007-09-04 2019-03-26 L'oreal Use of a lysate of bifidobacterium species for treating sensitive skin
CN110964656A (en) * 2018-09-30 2020-04-07 内蒙古伊利实业集团股份有限公司 Bifidobacterium lactis capable of preventing osteoporosis and application thereof
US10653728B2 (en) 2016-10-17 2020-05-19 New York University Probiotic compositions for improving metabolism and immunity
US10857090B2 (en) 2013-07-05 2020-12-08 Stellar Biome Inc. Oral compositions
KR102198552B1 (en) * 2020-06-09 2021-01-05 주식회사 비피도 Composition improving, preventing or treating halitosis or dental caries comprising Lactobacillus fermentum OK derived from human oral cavity
KR102198553B1 (en) * 2020-06-09 2021-01-05 주식회사 비피도 Composition improving, preventing or treating halitosis or dental caries comprising Lactobacillus gasseri HHuMIN D derived from human oral cavity
WO2021112041A1 (en) * 2019-12-02 2021-06-10 雪印メグミルク株式会社 Bone-strengthening agent
US11154731B2 (en) 2007-09-04 2021-10-26 L'oreal Cosmetic use of Bifidobacterium species lysate for the treatment of dryness
US11298382B2 (en) 2018-09-30 2022-04-12 Inner Mongolia Yili Industrial Group Co., Ltd. Bifidobacterium lactis BL-99 and application thereof
CN114711355A (en) * 2022-04-21 2022-07-08 暨南大学 Composite organic acid high-calcium probiotic drink taking astragalus polysaccharide as raw material as well as preparation method and application thereof
CN116904378A (en) * 2023-09-14 2023-10-20 山东中科嘉亿生物工程有限公司 Lactobacillus johnsonii JYLO-010 for promoting bone growth, microbial inoculum and application

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0273485A2 (en) * 1986-11-25 1988-07-06 Verenigde Coöperatieve Melkindustrie Coberco B.A. A process for preparing a raw material for human food; human food; a process for preparing a pharmaceutical composition
WO1997002757A1 (en) * 1995-07-10 1997-01-30 Merrick's, Inc. A feed fortifier and enhancer for preruminant

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0273485A2 (en) * 1986-11-25 1988-07-06 Verenigde Coöperatieve Melkindustrie Coberco B.A. A process for preparing a raw material for human food; human food; a process for preparing a pharmaceutical composition
WO1997002757A1 (en) * 1995-07-10 1997-01-30 Merrick's, Inc. A feed fortifier and enhancer for preruminant

Cited By (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7709537B1 (en) 1999-10-19 2010-05-04 Meiji Dairiese Corporation Food materials useful in preventing and ameliorating metabolic bone diseases and preventives/remedies for metabolic bone diseases comprising these materials
EP1230920A4 (en) * 1999-10-19 2004-09-29 Meiji Dairies Corp Food materials useful in preventing and ameliorating metabolic bone diseases and preventives/remedies for metabolic bone diseases comprising these materials
EP1230920A1 (en) * 1999-10-19 2002-08-14 Meiji Dairies Corporation Food materials useful in preventing and ameliorating metabolic bone diseases and preventives/remedies for metabolic bone diseases comprising these materials
JP2003300900A (en) * 2002-04-11 2003-10-21 Lotte Co Ltd Calcium absorption promoter, promoter of crystal growth of calcium phosphate, and food and drink and fodder containing them
US6872565B2 (en) 2003-01-29 2005-03-29 Biogaia Ab Product containing Lactobacillus reuteri strain ATTC PTA-4965 or PTA-4964 for inhibiting bacteria causing dental caries
US8658157B2 (en) 2003-01-29 2014-02-25 Biogaia Ab Selection and use of lactic acid bacteria for reducing dental caries and bacteria causing dental caries
US8343483B2 (en) 2003-01-29 2013-01-01 Biogaia Ab Selection and use of lactic acid bacteria for reducing dental caries and bacteria causing dental caries
US7517681B2 (en) 2003-01-29 2009-04-14 Biogaia Ab Selection and use of lactic acid bacteria for reducing dental caries and bacteria causing dental caries
JP2004315477A (en) * 2003-04-18 2004-11-11 Tokyoto Koreisha Kenkyu Fukushi Shinko Zaidan Bone resorption inhibitor
FR2872047A1 (en) * 2004-06-23 2005-12-30 Oreal COMPOSITION FOR SENSITIVE SKINS COMBINING MINERAL AND PROBIOTIC CATION (S)
WO2006000992A1 (en) * 2004-06-23 2006-01-05 L'oreal Method and compositions useful for preventing and/or treating sensitive and/or dry skin
EP1609463A1 (en) * 2004-06-23 2005-12-28 L'oreal Process and compositions for the protection and/or the treatment of dry and/or sensitive skin
US7651680B2 (en) 2004-06-23 2010-01-26 L'oreal Methods and compositions for preventing and treating sensitive and dry skin
US7956031B2 (en) 2005-05-31 2011-06-07 Naidu Lp Metallo-lactoferrin-coenzyme compositions for trigger and release of bioenergy
US9180148B2 (en) 2005-05-31 2015-11-10 Naidu Lp Metallo-lactoferrin-coenzyme compositions and methods for weight management
US9283253B2 (en) 2005-05-31 2016-03-15 Naidu Lp Metallo-lactoferrin-coenzyme compositions for detoxification support
US8476223B2 (en) 2005-05-31 2013-07-02 Naidu Lp Metallo-lactoferrin-coenzyme compositions to improve sleep patterns
US9687513B2 (en) 2005-07-05 2017-06-27 Probi Ab Methods of treating anemia and increasing the absorption of non-heme iron by administration of lactobacillus plantarum
US10105403B2 (en) 2005-07-05 2018-10-23 Probi Ab Methods of treating iron deficiency to prevent iron-deficiency anemia by administration of Lactobacillus plantarum
US8968726B2 (en) 2006-04-28 2015-03-03 Naidu Lp Lactoferrin and angiogenin compositions and uses thereof
US8021659B2 (en) * 2006-04-28 2011-09-20 Naidu Lp Coenzyme Q10, lactoferrin and angiogenin compositions and uses thereof
US10238897B2 (en) 2007-09-04 2019-03-26 L'oreal Use of a lysate of bifidobacterium species for treating sensitive skin
US11154731B2 (en) 2007-09-04 2021-10-26 L'oreal Cosmetic use of Bifidobacterium species lysate for the treatment of dryness
US9603876B2 (en) 2008-09-25 2017-03-28 New York University Compositions and methods for restoring gastrointestinal microbiota following antibiotic treatment
US8481299B2 (en) 2009-03-04 2013-07-09 L'oreal Use of probiotic microorganisms to limit skin irritation
EA022291B1 (en) * 2009-09-17 2015-12-30 Данон, С.А. Procedure for preparing pasteurised and fermented dairy product
CN102638994A (en) * 2009-09-17 2012-08-15 达能股份公司 Pasteurised and fermented dairy product and procedure for preparing such product
WO2011066880A2 (en) 2009-09-17 2011-06-09 Danone, S.A. Pasteurised and fermented dairy product and procedure for preparing such product
WO2011066880A3 (en) * 2009-09-17 2011-08-25 Danone, S.A. Pasteurised and fermented dairy product supplemented with calcium and vitamin d and procedure for preparing such product
ES2365145A1 (en) * 2009-09-17 2011-09-23 Danone, S.A. Fermented pasterised dairy product and preparation procedure of the same. (Machine-translation by Google Translate, not legally binding)
US8951512B2 (en) * 2010-05-04 2015-02-10 New York University Methods for treating bone disorders by characterizing and restoring mammalian bacterial microbiota
US20110280840A1 (en) * 2010-05-04 2011-11-17 New York University Compositions and methods for treating obesity and related disorders by characterizing and restoring mammalian bacterial microbiota
US9386793B2 (en) 2010-08-20 2016-07-12 New York University Compositions and methods for treating obesity and related disorders by characterizing and restoring mammalian bacterial microbiota
US10245290B2 (en) 2013-04-03 2019-04-02 Probi Ab Probiotic strains for use in treatment or prevention of osteoporosis
WO2014163568A1 (en) * 2013-04-03 2014-10-09 Probi Ab Probiotic strains for use in treatment or prevention of osteoporosis
AU2014250113B2 (en) * 2013-04-03 2017-03-30 Probi Ab Probiotic strains for use in treatment or prevention of osteoporosis
JP2016521123A (en) * 2013-04-03 2016-07-21 プロビ アーベ Probiotic strains used for the treatment or prevention of osteoporosis
CN105188723A (en) * 2013-04-03 2015-12-23 普罗比公司 Probiotic strains for use in treatment or prevention of osteoporosis
RU2636027C2 (en) * 2013-04-03 2017-11-17 Проби Аб Probiotic strains for application in osteoporosis treatment or prevention
CN104152354A (en) * 2013-05-14 2014-11-19 仙农生物科技(上海)有限公司 Compound probiotics as well as production process and application thereof to leavening agent
US10857090B2 (en) 2013-07-05 2020-12-08 Stellar Biome Inc. Oral compositions
CN104186655B (en) * 2014-08-27 2016-06-29 内蒙古伊利实业集团股份有限公司 There is low-fat yogurt of blood fat reducing and anti-oxidation function and preparation method thereof
CN104186655A (en) * 2014-08-27 2014-12-10 内蒙古伊利实业集团股份有限公司 Low-fat fermented milk having blood fat reduction and anti-oxidization functions and preparation method thereof
US10653728B2 (en) 2016-10-17 2020-05-19 New York University Probiotic compositions for improving metabolism and immunity
CN106858605A (en) * 2016-12-26 2017-06-20 吉林舒润生物科技有限公司 A kind of probiotics calcium functional food for preventing and treating bone information relevant disease
CN108245479A (en) * 2017-09-19 2018-07-06 王亚东 A kind of facial mask containing bifidobacterium lactis fermentation activity extract
CN108245479B (en) * 2017-09-19 2020-11-20 魏珂 Facial mask containing bifidobacterium lactis fermented active extract
CN110964656B (en) * 2018-09-30 2021-05-28 内蒙古伊利实业集团股份有限公司 Bifidobacterium lactis BL-99 capable of preventing osteoporosis and application thereof
CN110964656A (en) * 2018-09-30 2020-04-07 内蒙古伊利实业集团股份有限公司 Bifidobacterium lactis capable of preventing osteoporosis and application thereof
US11298382B2 (en) 2018-09-30 2022-04-12 Inner Mongolia Yili Industrial Group Co., Ltd. Bifidobacterium lactis BL-99 and application thereof
US11911422B2 (en) 2018-09-30 2024-02-27 Inner Mongolia Yili Industrial Group Co., Ltd. Bifidobacterium lactis BL-99 and application thereof
US11911423B2 (en) 2018-09-30 2024-02-27 Inner Mongolia Yili Industrial Group Co., Ltd. Bifidobacterium lactis BL-99 and application thereof
WO2021112041A1 (en) * 2019-12-02 2021-06-10 雪印メグミルク株式会社 Bone-strengthening agent
KR102198553B1 (en) * 2020-06-09 2021-01-05 주식회사 비피도 Composition improving, preventing or treating halitosis or dental caries comprising Lactobacillus gasseri HHuMIN D derived from human oral cavity
KR102198552B1 (en) * 2020-06-09 2021-01-05 주식회사 비피도 Composition improving, preventing or treating halitosis or dental caries comprising Lactobacillus fermentum OK derived from human oral cavity
CN114711355A (en) * 2022-04-21 2022-07-08 暨南大学 Composite organic acid high-calcium probiotic drink taking astragalus polysaccharide as raw material as well as preparation method and application thereof
CN116904378A (en) * 2023-09-14 2023-10-20 山东中科嘉亿生物工程有限公司 Lactobacillus johnsonii JYLO-010 for promoting bone growth, microbial inoculum and application
CN116904378B (en) * 2023-09-14 2023-12-29 山东中科嘉亿生物工程有限公司 Lactobacillus johnsonii JYLO-010 for promoting bone growth, microbial inoculum and application

Also Published As

Publication number Publication date
AU6838500A (en) 2001-03-26
WO2001015715A3 (en) 2001-10-18
GB9920578D0 (en) 1999-11-03

Similar Documents

Publication Publication Date Title
WO2001015715A2 (en) Composition for maintenance of bone or dental health or treatment of bone or dental disorders
US10105403B2 (en) Methods of treating iron deficiency to prevent iron-deficiency anemia by administration of Lactobacillus plantarum
Adolfsson et al. Yogurt and gut function
Suvarna et al. Probiotics in human health: A current assessment
Mitsuoka Development of functional foods
US7150986B2 (en) Lactic acid bacteria-containing probiotics products
EP3375448B1 (en) Strain having ability to inhibit obesity and pharmaceutical composition containing same
Umer Khan Probiotics in dairy foods: a review
JPWO2007043563A1 (en) Composition for improving intestinal flora
Chandan Role of milk and dairy foods in nutrition and health
MXPA01006822A (en) Milk protein hydrolysate for addressing a bone or dental disorder
JP2023011739A (en) Fermented milk for promoting elevation of amino acid concentration in blood
JP2023175938A (en) Glucose metabolism improving composition
WO2016020485A1 (en) A combination of vitamin d and zinc and its use
Pato et al. Hypocholesterolemic effect of indigenous dadih lactic acid bacteria by deconjugation of bile salts
WO2010061877A1 (en) Mineral absorption improver, and method for improving absorption of minerals
CN111201026B (en) Fermented milk and polysaccharides with cancer cachexia inhibiting effect
EP2419506B1 (en) Bacterial strains having an outstanding ability to produce menaquinone
Campbell et al. Yogurt consumption does not enhance immune function in healthy premenopausal women
Ahmad et al. Probiotic potential of lactic acid to improve immunity during COVID‐19 pandemic (a mini review)
US20230149482A1 (en) Composition for improving gut microbiota
EP3177278A1 (en) Vitamin b2 and its use
JP2020014452A (en) Method for producing concentrated fermented milk
Chandan Health benefits of yogurt
WO2021117857A1 (en) Composition for lowering heart rate

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
AK Designated states

Kind code of ref document: A3

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase in:

Ref country code: JP