WO2019166985A1

WO2019166985A1 - Probiotic composition

Info

Publication number: WO2019166985A1
Application number: PCT/IB2019/051620
Authority: WO
Inventors: Karin POSTAL
Original assignee: Propharma Semplificata Srl
Current assignee: Propharma Semplificata Srl
Priority date: 2018-02-28
Filing date: 2019-02-28
Publication date: 2019-09-06
Anticipated expiration: 2020-08-28
Also published as: IT201800003128A1

Abstract

The probiotic composition comprises at least one microorganism belonging to the species Saccharomyces Boulardii and α-galacto-oligosaccharides.

Description

PROBIOTIC COMPOSITION

Technical Field

The present invention relates to a probiotic composition.

Background Art

It is well known how attention to food and diet are among the most important issues for the population today.

Often, the lack of time associated with hectic lifestyles leaves no room for the search for healthy and nutritious foods.

In addition, nutraceuticals are becoming increasingly important for the production of food produces that have both a nutritional and beneficial function for the body.

In this regard, it is well known that, in many industrial sectors, there is a particularly strong need to renew products and to look for new ones in order to distinguish these more clearly from those of the competition.

In this scenario, diets for the elderly and children are particularly important, as both have diets whose regulation is of fundamental importance for their psycho physical well-being.

With particular reference to the elderly, it should be stressed that the diet of the latter provides for a low-calorie need due to the slowdown in the basal metabolism and a decrease in skeletal muscles and physical activity.

However, despite the reduced calorie requirements necessary to sustain the elderly, the latter need a diet that includes tasty and easily ingestible foods which arouse curiosity and appetite in the same.

In addition, the condition of physiological immunosuppression to which elderly people are subjected makes their intestinal flora particularly sensitive to chemical-physical changes causing them numerous disorders and discomforts, including diarrhea, colitis and gastric illnesses.

Furthermore, these categories of patients are often subject to the administration of antibiotics, frequent cause of intestinal infections and diarrheas.

In the face of such clinical situations, probiotic compositions comprising predefined mixtures of microorganisms producing antimicrobial substances such as bacteriocins, lactic acid, hydrogen peroxide and proteases are currently in use and are adapted to compete with pathogenic microorganisms present in the anatomical area affected by infections, such as the intestine or stomach. Nevertheless, these compositions have many drawbacks including the fact that, once they have been ingested and have reached the anatomical area of interest, the microorganisms present in them lose their vitality, and therefore their effectiveness.

In fact, known compositions have a reduced resistance to digestive processes, and in particular to the pH reached during these processes; this means that the degradation of the compositions prevents overcoming the gastric barrier and results in a significant decrease in the effectiveness of the same.

Moreover, the antimicrobial substances produced by the microorganisms are hydrolysable by the human intestine and prevent the latter from reaching the colon.

A further drawback is tied to the fact that these probiotic compositions must be administered to patients before or after meals, and always separately.

It is easy to understand how in the case of patients who undergo numerous daily drug administrations, the fact of having to submit them to a further administration of these compositions has a considerable impact on the perseverance of the treatment, significantly reducing its effectiveness.

In addition, the known compositions are difficult to mix with foods such as ice creams, fruit juices, mousses or the like, because of their low resistance to the chemical-physical variations of the surrounding environment.

In this case too, in fact, the microorganisms present in the known compositions lose their respective vitality, cancelling the effectiveness of the compositions themselves.

Other compositions of known type are described in patent documents no. WO 02/060276, WO 2013/001089, WO 2010/081126, WO 2007/125558, EP 2 438 821 and US 2015/044331. Another type of composition is known from the Mintel document“Sugar Free Probiotic Medium Fat Ice Cream”.

In particular, patent document no. WO 2007/125558 describes a probiotic composition the use of which is strongly limited to non-elderly patients. This composition has, in fact, a reduced efficacy from the metabolic point of view. In addition, the composition described in WO 02/060276 has numerous drawbacks, including the fact that it has a significantly limited spectrum of effectiveness, especially in patients undergoing antibiotic therapy.

Description of the Invention

The main aim of the present invention is to provide a probiotic composition that is resistant to the digestive processes and the relative pH reached during these processes, allowing the overcoming of the gastric barrier and enabling the antimicrobial substances produced by the microorganisms themselves to reach the colon.

Another object of the present invention is to provide a probiotic composition in which the microorganisms present prolonged vitality with respect to known compositions.

A further object of the present invention is to provide a probiotic composition for use in the treatment of diarrhea due to antibiotics, intestinal infections and traveler’ s diarrhea.

Yet another object of the present invention is to provide a probiotic composition that can be mixed with food, thus increasing the perseverance of the administration of the same by patients.

Another object of the present invention is to provide a probiotic composition that allows overcoming the aforementioned drawbacks of the prior art in a simple, rational, easy, effective to use and low-cost solution.

The aforementioned objects are achieved by the present probiotic composition having the characteristics of claim 1.

Brief Description of the Drawings

Other characteristics and advantages of the present invention will be more evident from the description of a preferred, but not exclusive, embodiment of a probiotic composition, illustrated by way of an indicative, but non-limiting example, in the attached tables of drawings in which:

Figure 1 is a graph representing the survival trend of the microorganisms present in the composition according to the invention at different storage times; Figure 2 is a graph representing the survival trend of Lactobacillus Casei 431 in the composition according to the invention when used for the formulation of an ice cream;

Figure 3 is a graph representing the survival trend of Bifidobacterium animalis subsp. lactis Bbl2 in the composition according to the invention when used for the formulation of an ice cream;

Figure 4 is a graph representing the survival trend of Saccharomyces Boulardii in the composition according to the invention when used for the formulation of an ice cream.

Embodiments of the Invention

In the present treatise reference is made to a probiotic composition.

This probiotic composition comprises one microorganism belonging to the species Saccharomyces Boulardii and a-galacto-oligosaccharides.

The microorganism belonging to the species Saccharomyces Boulardii is present in a concentration by weight, evaluated with respect to the total weight of the composition, higher than 0.05% and preferably comprised between 0.1% and 5%.

The a-galacto-oligosaccharides are composed of a glucose unit linked to 1, 2, or 3 galactose molecules by means of a-glycosidic bonds.

The a-galacto-oligosaccharides are preferably present in a concentration by weight, evaluated with respect to the total weight of the composition, higher than 50% and preferably comprised between 70% and 99%.

Furthermore, the composition comprises at least one microorganism selected from the group comprising: Lactobacillus Casei 431, Lactobacillus Rhamnosus GG, Lactobacillus Reuteri ATTC 5530 and Bifidobacterium animalis subsp. lactis Bbl2.

In particular, in accordance with a preferred embodiment of the composition, the latter comprises Lactobacillus Casei 431 present in a concentration by weight, evaluated with respect to the total weight of the composition, lower than 0.5% and preferably comprised between 0.01 and 3%.

Furthermore, the composition preferably comprises Bifidobacterium animalis subsp. lactis Bbl2 present in a concentration by weight, evaluated with respect to the total weight of the composition, lower than 0.5% and comprised between 0.01% and 0.5%. The aforementioned composition is in powdery form.

In other words, the composition according to the present invention consists of a powder.

Alternative embodiments cannot however be ruled out wherein the composition is in liquid or gel form.

Furthermore, the present invention provides a composition according to the invention, for use in the treatment and/or prevention of diarrhea due to antibiotics, intestinal infections, traveler’s diarrhea, ulcerative colitis, Crohn’s disease, irritable bowel syndrome and atopic dermatitis.

It should be noticed that the synergistic combination of a-galacto- oligosaccharides with Lactobacillus Casei 431, Bifidobacterium animalis subsp. lactis Bbl2 and Saccharomyces Boulardii allows the colonization of the human intestine and adherence to cells of the intestinal mucosa achieving a trophic effect on the latter.

In addition the present invention relates to the use of the composition in liquid or semisolid food preparations.

More specifically, food preparations are selected from the group comprising: ice cream, fruit puree, vegetable puree, fruit juice, yoghurt, mousse, smoothie, extracts and soluble foods.

Preferably, these food preparations are freeze-dried.

For the purposes of the present discussion, the words“soluble products” mean products which are adapted to be dissolved in a solvent phase, such as e.g. chamomile, tea and the like.

In this regard, the present invention also relates to a probiotic preparation for food preparations comprising the composition according to the present invention and a basic component, in turn, comprising a mixture of a thickening agent and an emulsifying agent.

Preferably, the emulsifying agent is selected from the list comprising: curdlan, glucomannan, polyoxyethylene stearate, polyoxyethylene sorbitol monolaurate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitol monopalmitate, polyoxyethylene sorbitol monostearate, polyoxyethylene sorbitan tristearate, pectin, gelatin, sucrose acetate isobutyrate, glycerol esters of resin, disodium diphosphate, trisodium diphosphate, tetrasodium diphosphate, tetrapotassium diphosphate, diphosphate di-calcium, calcium hydrogen diphosphate, pentasodium triphosphate, pentapotassium triphosphate, sodium polyphosphates, potassium polyphosphates, sodium and calcium polyphosphates, calcium polyphosphates, b-cyclodextrin, microcrystalline cellulose, cellulose powder, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, enzymatically hydrolyzed carboxymethyl cellulose, salts of sodium, potassium and calcium of fatty acids, salts of magnesium of fatty acids, mono- and diglycerides of fatty acids, acetic esters of mono- and diglycerides of fatty acids, lactic esters of mono- and diglycerides of fatty acids, citric esters of mono- and diglycerides of fatty acids, tartaric esters of mono- and diglycerides of fatty acids, mono- and diacetyl tartarartic esters of mono- and diglycerides of fatty acids, mixed acetic and tartaric esters of mono- and diglycerides of fatty acids, saccharic esters of fatty acids, saccharo-glycerides, polyglyceric esters of fatty acids, polyglyceryl polyricinoleate, propane- l,2-diol esters of fatty acids, propylene glycol esters of fatty acids, lactylate glyceryl- and 1 -propyl esters of fatty acids, soybean oil thermally oxidized with mono- and diglycerides of fatty acids, stearoyl 2- lactylate, sodium salt, stearoyl lactylate, calcium salt, stearyl tartrate, sorbitol monostearate, sorbitol tristearate, sorbitol monolaurate, sorbitol monooleate, sorbitol monopalmitate.

At the same time, the thickening agent is selected from the list comprising: alginic acid, sodium alginate, potassium alginate, ammonium alginate, calcium alginate, propane- l,2-diol alginate, agar-agar, carrageenan, processed Eucheuma seaweed, carob seed flour, guar gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, gellan gum.

Preferably, the a-galacto-oligosaccharides, Saccharomyces Boulardii, Bifidobacterium animalis subsp. lactis Bbl2, Lactobacillus Casei 431 and the basic component are present in the following concentrations by weight, evaluated with respect to the total weight of the preparation: a-galacto-oligosaccharides 15% - 30%

Saccharomyces Boulardii 0,01% - 0,5% Bifidobacterium animalis subsp. lactis Bbl2 0,01% - 0,5% Lactobacillus Casei 431 0,01% - 0,5% Basic component 60% - 85%.

An example of such a preparation is given below.

EXAMPLE 1

A food preparation, preferably ice cream, has been produced, comprising a- galacto-oligosaccharides, Saccharomyces Boulardii, Lactobacillus Casei 431 and a basic component, present in the following concentrations by weight:

a-galacto-oligosaccharides 23.16%

Bifidobacterium animalis subsp. lactis Bbl2 0.046%

Lactobacillus Casei 431 0.046%

Saccharomyces Boulardii 0.46% basic component 77.0%

In detail, the composition according to the invention has the following concentration of microorganisms:

Bifidobacterium animalis subsp. lactis Bbl2 2.3xl0⁸ CFU

Lactobacillus Casei 431 1.4X10⁸ CFU

Saccharomyces Boulardii l.4Xl0⁸ CFU

According to the composition of the example 1, the basic component comprises two thickening agents and three emulsifying agents.

More specifically, the thickening agents are guar gum flour and carob seed gum, while the emulsifying agents are mono- and diglycerides of fatty acids, mono- and diglyceric esters of fatty acids, and propylene glycol esters of fatty acids.

Preferably, the mixture also comprises fructose, skimmed milk powder and starch.

The formulation shown in example 1 is used in the preparation of ice-cream- type foods. In fact, the fact of mixing the composition comprising the a-galacto- oligosaccharides, Saccharomyces Boulardii, Lactobacillus Casei 431 and the basic component according to example 1 allows it to be mixed cold.

This means that the aforementioned preparation can be used in the preparation of ice cream at room temperature.

A comparative viability test of the microorganisms present in the composition according to the invention and in the ice cream obtained from the latter was therefore carried out.

The viability of each microorganism was assessed on the individual strains. In particular, a colony count was performed on agarose plates.

The following selective media were used:

- Bifidobacterium animalis subsp. lactis Bbl2 Beeren’s Agar

- Lactobacillus Casei 431 MRS + vancomycin agar

- Saccharomyces Boulardii YPD agar

The composition shown in Example 1 below has been aliquotted in portions containing lOOg each and stored at 4°C under vacuum.

For each viability test, a sealed and open aliquot was used to perform the test at 0, 1, 3 and 6 months after preparation.

At the same time, the composition according to the Example 1 was used for the preparation of 500g of ice cream. On the latter, the tests were performed at 0, 1, 2 and 3 days from preparation.

Three technical replicas and three experimental replicas were performed on each test.

The count of the colonies was performed on the respective selective media. The data are provided below relating to the viability trend over time.

Table 1

Table 1 and Figures 1 and 2 show the viability trend over time of the microorganisms of the composition according to the invention.

From the aforementioned analysis it can be observed that the viability of the microbial strains remains unexpectedly unchanged until 3 days after preparation of the composition according to the invention, significantly increasing the effectiveness of the composition.

It has in practice been ascertained that the described invention achieves the intended objects.

The fact is underlined that the particular solution of providing the synergistic combination of a-galacto-oligosaccharides with Lactobacillus Casei 431, Bifidobacterium animalis subsp. lactis Bbl2 and Saccharomyces Boulardii permits significantly increasing bifidogenic activity, compared to known compositions, and permitting the growth of bifidobacteria and lactobacilli in the intestine.

In addition, the particular solution of providing a-galacto-oligosaccharides allows obtaining a composition which is not hydrolysable by human digestive enzymes, preventing the absorption of the a-galacto-oligosaccharides themselves by the intestinal mucosa; this allows fermentation at the level of the colon where the latter undergo fermentation phenomena by the intestinal microflora, resulting in the increase of bifidobacteria with a consequent improvement in blood sugar levels.

Furthermore, the fact of providing the synergistic combination of a-galacto- oligosaccharides with Lactobacillus Casei 431, Bifidobacterium animalis subsp. lactis Bbl2 and Saccharomyces Boulardii permits considerably increasing the absorption of minerals, bringing considerable benefits to the improvement of bone density.

Finally, the particular solution of providing the aforementioned combination of a-galacto-oligosaccharides, Lactobacillus Casei 431, Bifidobacterium animalis subsp. lactis Bbl2 and Saccharomyces Boulardii with a basic component allows obtaining foods with low sweetening power.

Claims

1) Probiotic composition, characterized by the fact that it comprises at least one microorganism belonging to the species Saccharomyces Boulardii and a- galacto-oligosaccharides.

2) Composition according to claim 1, characterized by the fact that said a- galacto-oligosaccharides are present in a concentration by weight, evaluated with respect to the total weight of the composition, higher than 50% and preferably comprised between 70% and 99%.

3) Composition according to one or more of the preceding claims, characterized by the fact that said microorganism belonging to the species Saccharomyces Boulardii is present in a concentration by weight, evaluated with respect to the total weight of the composition, higher than 0.05% and preferably comprised between 0.1% and 5%.

4) Composition according to one or more of the preceding claims, characterized by the fact that it comprises at least one microorganism selected from the group comprising: Lactobacillus Casei 431, Lactobacillus Rhamnosus GG, Lactobacillus Reuteri ATTC 5530 and Bifidobacterium animalis subsp. lactis Bbl2.

5) Composition according to one or more of the preceding claims, characterized by the fact that it comprises said Lactobacillus Casei 431 present in a concentration by weight, evaluated with respect to the total weight of the composition, lower than 0.5% and preferably comprised between 0.01 and 0.3%.

6) Composition according to one or more of the preceding claims, characterized by the fact that it comprises said Bifidobacterium animalis subsp. lactis Bbl2 present in a concentration by weight, evaluated with respect to the total weight of the composition, lower than 0.5% and preferably comprised between 0.01 and 0.3%.

7) Composition according to one or more of the preceding claims, characterized by the fact that it is in powdery form.

8) Composition according to one or more of the preceding claims, for use in the treatment and/or prevention of antibiotic diarrhea, intestinal infections, traveler's diarrhea, ulcerative colitis, Crohn's disease, irritable bowel syndrome and atopic dermatitis.

9) Use of the composition according to one or more of the preceding claims, in at least one of solid, liquid and semisolid food preparations.

10) Use of the composition according to one or more of the preceding claims, in which said food preparations are selected from the list comprising: ice cream, fruit puree, vegetable puree, fruit juice, yoghurt, mousse, smoothie, extracts and soluble foods.

11) Use of the composition according to one or more of the preceding claims, in which said food preparations are freeze-dried.

12) Probiotic preparation for foods, characterized by the fact that it comprises the composition according to one or more of claims 1 to 8 and at least one basic component, in turn, comprising a mixture of at least one thickening agent and at least one emulsifying agent.

13) Preparation according to claim 10, characterized by the fact that said emulsifying agent is selected from the list comprising: curdlan, glucomannan, polyoxyethylene stearate, polyoxyethylene sorbitol monolaurate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitol monopalmitate, polyoxyethylene sorbitol monostearate, polyoxyethylene sorbitan tristearate, pectin, gelatin, sucrose acetate isobutyrate, glycerol esters of resin, disodium diphosphate, trisodium diphosphate, tetrasodium diphosphate, tetrapotassium diphosphate, diphosphate di-calcium, calcium hydrogen diphosphate, pentasodium triphosphate, pentapotassium triphosphate, sodium polyphosphates, potassium polyphosphates, sodium and calcium polyphosphates, calcium polyphosphates, b-cyclodextrin, microcrystalline cellulose, cellulose powder, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, enzymatically hydrolyzed carboxymethyl cellulose, salts of sodium, potassium and calcium of fatty acids, salts of magnesium of fatty acids, mono- and diglycerides of fatty acids, acetic esters of mono- and diglycerides of fatty acids, lactic esters of mono- and diglycerides of fatty acids, citric esters of mono- and diglycerides of fatty acids, tartaric esters of mono- and diglycerides of fatty acids, mono- and diacetyl tartarartic esters of mono- and diglycerides of fatty acids, mixed acetic and tartaric esters of mono- and diglycerides of fatty acids, saccharic esters of fatty acids, saccharo-glycerides, polyglyceric esters of fatty acids, polyglyceryl polyricinoleate, propane- l,2-diol esters of fatty acids, propylene glycol esters of fatty acids, lactylate glyceryl- and 1 -propyl esters of fatty acids, soybean oil thermally oxidized with mono- and diglycerides of fatty acids, stearoyl 2- lactylate, sodium salt, stearoyl lactylate, calcium salt, stearyl tartrate, sorbitol monostearate, sorbitol tristearate, sorbitol monolaurate, sorbitol monooleate, sorbitol monopalmitate, and said thickening agent is selected from the list comprising: alginic acid, sodium alginate, potassium alginate, ammonium alginate, calcium alginate, propane- l,2-diol alginate, agar-agar, carrageenan, processed Eucheuma seaweed, carob seed flour, guar gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, gellan gum.