WO2017009313A1

WO2017009313A1 - A tablet with a high concentration of lactic acid bacteria

Info

Publication number: WO2017009313A1
Application number: PCT/EP2016/066477
Authority: WO
Inventors: Franco Pirovano; Francesco Palladini
Original assignee: Ergon S.R.L.
Priority date: 2015-07-14
Filing date: 2016-07-12
Publication date: 2017-01-19
Also published as: ITUB20152151A1

Abstract

One object of the present invention is a tablet, which may be chewable, containing a high concentration of lactic acid bacteria. The tablet may be of limited size and the formulation has been found to be stable beyond the usual storage times typical of lactic acid bacteria formulations kept at ambient temperature.

Description

A TABLET WITH A HIGH CONCENTRATION OF LACTIC ACID BACTERIA

The present invention relates to the field of compositions for food and pharmaceutical use, and particularly to the formulation of tablets containing lactic acid bacteria.

Background of the invention

Lactic acid bacteria are used not only in the production of dairy products, but also in products known as food supplements or pharmaceutical products.

Lactic acid bacteria contribute to the maintenance of correct physiology of the intestinal system, and are now also used in the treatment of disorders of the digestive system and even recognized pathologies. Some selected strains of lactic acid bacteria are also used in the treatment of the female urogenital system.

Because of this increasingly widespread use, lactic acid bacteria are formulated in compositions based on pharmaceutical technology, and the products are known as "food supplements". The proven efficacy of lactic acid bacteria has led medical practitioners to prescribe them with increasing frequency.

The general public have also become aware of the beneficial effect of lactic acid bacteria, and they buy lactic acid bacteria increasingly commonly in order to increase or maintain well-being, or even by way of self-medication.

Lactic acid bacteria are sensitive to environmental conditions, and their survival and consequent efficacy depends on special technological arrangements for their formulation.

Normally, lactic acid bacteria are formulated in freeze-dried form, in order to prolong their life. The formulations may be in different forms, for example sachets and capsules, but are also formulated as suppositories, vaginal suppositories and ovules.

The efficacy of the lactic acid bacteria formulation depends on the amount of bacteria present in the dosing unit.

In order to obtain high concentrations of lactic acid bacteria per dosing unit, the whole of the formulation, including the excipients, reaches a weight such that it is necessary to use high dosing units, requiring the use of the sachet or capsule form.

In many cases, especially where high-concentration formulations are concerned, the product must be kept at low temperature, typically at domestic refrigerator temperature or below. If the product has to be taken outside, for the purpose of travel for example, the viability, and therefore the efficacy, of lactic acid bacteria decreases rapidly with time.

Capsule formulations kept at ambient temperature usually contain lower concentrations, but the viability of the bacteria is still time-limited and temperature-dependent. It is known that lactic acid bacteria are fragile organisms, and attempts to formulate tablets with a content of lactic acid bacteria encountered the problem of high mortality during compression.

WO2008/039561 describes a formulation of lactic acid bacteria in which viability is increased by the inclusion in the formulation of a carrier comprising at least two prebiotic carbohydrate alcohols selected from the group consisting of erythritol, lactitol, mannitol, sorbitol and xylitol. A specific example present in this document is a mixture of mannitol and sorbitol, known by the commercial name of Pharmaburst^®.

WO2010054439 describes a formulation in which the probiotic is included in a matrix, preferably anhydrous. The amount of viable lactic acid bacteria is not greater than 10⁹ CFU/g.

One of the most common uses of lactic acid bacteria is the treatment of transient intestinal disorders such as diarrhoea. Consequently lactic acid bacteria formulations are increasingly widely used by travellers, especially in the tropics.

As mentioned above, the efficacy of the formulation depends directly on the concentration of live acid bacteria, among other factors.

This leads to a first problem concerning formulation. In order to achieve a high concentration of lactic acid bacteria per dosing unit of at least 10⁹, or commonly at least 10¹² CFU/g of formulation, sachets or capsules must be used. In addition to the problem of storage at temperatures which may be high, in the tropics for example, and where refrigerated storage is often unavailable, there is also a problem of consumption. The sachet requires a liquid medium, such as water or a beverage, in which to suspend the dose to be taken, whereas, in some countries, water is not immediately accessible, or some beverages are incompatible with lactic acid bacteria. Tablets, because of their size, are subject to the same problems of consumption as sachets, with further possible difficulties for children, elderly people, or persons who have difficulty in swallowing.

Consequently there is a need to provide a high-concentration formulation of lactic acid bacteria in the form of a chewable tablet, which has limited dimensions, and which is stable over time even at ambient temperature, particularly at high temperatures.

It is also known that lactic acid bacteria often have an unpleasant flavour, making it difficult to administer the formulations to children.

Summary of the invention

A tablet, which may be chewable, containing a high concentration of lactic acid bacteria, has now been found and is the object of the present invention. The tablet may be of limited size and the formulation has been found to be stable beyond the usual storage times typical of lactic acid bacteria formulations kept at ambient temperature. Surprisingly, it has been found that, with a particular formulation of excipients, it is possible to prepare a tablet containing a final concentration of up to 250x10⁹ CFU/g of lactic acid bacteria, while also greatly reducing mortality during compression.

Detailed description of the invention

One object of the present invention is a tablet containing lactic acid bacteria up to 250x 10⁹ CFU/g, comprising:

Lactic acid bacteria 1-70% w/w

Sorbitol 25-50% w/w

Magnesium stearate Up to 1-3% w/w

Talc Up to 1 % w/w

Silica Up to 1 % w/w

Flavouring agent Up to 1 % w/w

Sucralose Up to 1 % w/w

Hydroxyethyl cellulose Up to 4% w/w

In a preferred embodiment of the invention, the tablet has a final weight in the range from about 250 to about 300 mg, and contains up to about 62.5x10⁹ to about 75.0x 10⁹ per tablet.

In another aspect of the present invention, it has been found that strawberry, cream, and sweet/morello cherry flavourings effectively mask the flavour of lactic acid bacteria, which is not always pleasant, without adversely affecting their viability. Example of commercially available flavouring agent is cream flavour powder 1013439.

The tablet according to the present invention may contain a conventional sweetener, for example acesulfame K, stevia rebaudiana, or sucralose, the last-mentioned being preferred.

In a preferred embodiment of the present invention, a 250 mg tablet having the following composition is provided:

Sorbitol for direct compression 38.61

Klucel nutra D 4.71

AEROSIL 200 Pharma 0.94

Aerated talc 0.94

Sucralose 0.08

Cream flavour powder 1013439 0.80

Probiotics 52.00

Magnesium stearate 1.92

100.0000 A further advantage of the tablet according to the present invention is that it dissolves in the mouth, making the product suitable for persons who have difficulty in swallowing, for example children and elderly people, in spite of its limited dimensions.

The lactic acid bacteria that can be used in the present invention are all those accepted for human use, without any particular limitations.

The preferred lactic acid bacteria are those, in the form of either generic species or specific strains, alone or in a mixture, which are used for the treatment of intestinal disorders, particularly diarrhoea.

Other preferred lactic acid bacteria are those used in the treatment of more serious intestinal disorders.

Lactic acid bacteria used for oral administration for the treatment of disorders of the female urogenital system are also preferred.

Examples of lactic acid bacteria suitable for the present invention are Lactobacillus acidophilus, Lactobacillus buchneri, Lactobacillus casei, Lactobacillus catenaforme, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus cellobiosus, Lactobacillus jensenii, Lactobacillus delbrueckii, Lactobacillus leichmanii, Lactobacillus plantarum, Lactobacillus minutus, Lactobacillus salivarius, Bifidobacterium, Lactobacillus rogosae, Bifidobacterium adolescentis. Bifidobacterium angulatum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium dentium, Bifidobacterium eriksonii, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium plantarum, Bifidobacterium pseudo-catenulatum, Bifidobacterium pseudolongum, Streptococcus lactis, Streptococcus raffinolactis, Streptococcus thermophilus, Acidaminococcus fermenta, Cytophaga fermentans. Rhodoferax fermentans, Cellulomonas fermentans and Zymomanas mobilis, and mixtures thereof.

In the embodiment of the present invention relating to a tablet containing lactic acid bacteria indicated for the treatment of the female urogenital tract, examples of suitable lactic acid bacteria are: Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus brevis, Lactobacillus jensenii Lactobacillus casei, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus plantarum, Lactobacillus salivarius, subsp. salicinius and mixtures thereof.

The present invention further comprises specific strains.

In a preferred embodiment of the invention, the lactic acid bacteria Lactobacillus rhamnosus, Bifidobacterium breve, Lactobacillus plantarum, and Streptococcus thermophilus are used, individually or in mixtures of two or more of these.

In a more preferred embodiment of the invention, the following lactic acid bacteria strains were used: Lactobacillus rhamnosus LRH 28, Bifidobacterium breve Bbr 8, Lactobacillus plantarum 14D, and Streptococcus thermophilus Z57 were used, individually or in mixtures of two or more of these. The individual strains were deposited at the following: Belgian Coordinated Collections of Microorganism Nr. LMG 27341 (LRH 28); LMG P-17501 (Bbr8); LMG P-21908 (Z57); Coleccion espanola de cultivos tipo nr. CECT 4528 (14D).

In an even more preferred embodiment of the present invention, the tablet comprises a strain selected from the group consisting of Lactobacillus rhamnosus LRH 28, Bifidobacterium breve Bbr 8, Lactobacillus plantarum 14D, Streptococcus thermophilus Z57 and mixtures thereof.

In one embodiment of the present invention, the tablet contains a mixture of the following strains:

Lactobacillus rhamnosus LRH 28 9% (w/w)

Bifidobacterium breve Bbr 8 50% (w/w)

Lactobacillus plantarum 14D 18% (w/w)

Streptococcus thermophilus Z57 18% (w/w).

All the materials, particularly the excipients used in the preparation of the tablet according to the present invention, are commonly used in the industry and are of food and/or pharmaceutical grade.

Conveniently, the tablets according to the present invention are packaged in controlled-moisture systems.

In a preferred embodiment of the invention, the tablets are packaged in a moisture-controlled container, typically a container comprising a moisture absorber. An example applicable to the present invention is the container marketed by CSP Technologies under the brand Active-Vials^®.

Another object of the present invention is a method for manufacturing the aforesaid tablet.

The tablet according to the present invention is prepared by direct cold tableting. The tableting is of the very low compression type, typically 2 t/cm². The initial quantity of lactic acid bacteria for tableting is in the range from about 500* 10⁹ to about 600*10⁹ CFU/g, and the proportion relative to the excipients is as stated above, preferably 52:48 respectively (by weight).

Although there is a degree of mortality during compression, the tablet according to the present invention shows the unexpected advantage of prolonged storage. The tablet according to the present invention was subjected to stability tests, with the following results: Flavouring agents tested

Black cherry - Strawberry - Cream

(billions/g)

4°C 25°C 30°C 42°C

to 165

1 month 150 150 140 125

3 months 145 145 133 85

6 months 146 145 133 72

9 months 146 140 1 15 60

As can be seen, the present invention provides a tablet containing a very high concentration of lactic acid bacteria, although the compression method is not known in the prior art as the best method for formulations with a high concentration of viable lactic acid bacteria.

Of particular importance is the result showing stability at temperatures above the usual ambient temperature, making the tablet of the present invention particularly suitable for consumers travelling to hot countries, particularly those countries where traveller's diarrhoea is a problem.

The following examples further illustrate the invention. Example 1

STEP 1 : Weigh the sorbitol for direct compression, and add Klucel Nutra D, AEROSIL 200 Pharma, Aerated talc, Sucralose and cream flavour powder to a double-cone mixer. Add the 2P probiotic mix, ensure that the temperature of the clean room is 22°C and the humidity less than 30%. Mix for 30 minutes. STEP 2: Load the magnesium stearate into the double-cone mixer and mix for 5 minutes. STEP 3: Discharge the mixture, divide it into dedicated 10 kg bags and heat-seal the bags, place everything in the appropriate cold storage room at 5/8°C.

STEP 4: Fit the Ronchi 18/21 tablet press with 7 mm cylindrical punches, load the precompression spring at 10 to 30 kilonewton (10000 - 30000 newton) on the scale ranges. Pour the pre-mixed lactic acid bacteria powder into the forced loading hopper and start tableting at a rate of 20000 tablets per hour. Measure the physical parameters: with the Farmalabor Tech hardness tester, the tablet should break at 10 daN (1 daN = 10 N), the diameter of the tablet is 7 mm, and the height of the tablet is 5 mm.

Example 2

Using the procedure of Example 1 , a 250 mg tablet was prepared with the following composition:

Sorbitol for direct compression 38.6096

Klucel nutra D 4.7120

AEROSIL 200 Pharma 0.9424

Aerated talc 0.9424

Sucralose 0.0800

Cream flavour powder 0.8000

Probiotics 52.0000

Magnesium stearate 1.9136

100.0000 Example 3

Using the formulation of Example 2, a 250 mg tablet was prepared with the following composition of lactic acid bacteria:

Lactobacillus rhamnosus LRH 28 9% (w/w)

Bifidobacterium breve Bbr 8 50% (w/w) Lactobacillus plantarum 14D 18% (w/w)

Streptococcus thermophilus Z57 18% (w/w).

Example 4

The tablets prepared in the preceding examples were packaged in Active-Vials^® containers made by CSP Technologies.

Claims

1. A tablet comprising lactic acid bacteria at a concentration of up to 250x10 CFU/g, characterized in that it comprises:

a. Lactic acid bacteria 1-70% w/w;

b. Sorbitol 25-50% w/w.

2. Tablet according to claim 1 , having a weight of between 250 and 300 mg.

3. Tablet according to claim 1 or 2, having the following composition:

Lactic acid bacteria 1-70% w/w

Sorbitol 25-50% w/w

Magnesium stearate Up to 1-3% w/w

Talc Up to 1 % w/w

Silica Up to 1 % w/w

Flavouring agent Up to 1 % w/w

Sucralose Up to 1 % w/w

Hydroxyethyl cellulose Up to 4% w/w

4. Tablet according to claim 3, having the following composition:

Sorbitol for direct compression 38.61

Klucel nutra D 4.71

AEROSIL 200 Pharma 0.94

Aerated talc 0.94

Sucralose 0.08

Cream flavour powder 0.80

Probiotics 52.00

Magnesium stearate 1.92

5. Tablet according to any one of claims 1-4, wherein said lactic acid bacteria is selected from the group consisting of Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus casei, Lactobacillus catenaforme, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus delbrueckii, Lactobacillus leichmanii, Lactobacillus plantarum, Lactobacillus minutus, Lactobacillus salivarius, Bifidobacterium, Lactobacillus rogosae, Lactobacillus plantarum, Lactobacillus salivarius, subsp. Salicinius, Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium dentium, Bifidobacterium eriksonii, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium plantarum, Bifidobacterium pseudo- catenulatum. Bifidobacterium pseudolongum, Streptococcus lactis, Streptococcus raffinolactis, Streptococcus thermophilus, Acidaminococcus fermenta, Cytophaga fermentans, Rhodoferax fermentans, Cellulomonas fermentans and Zymomanas mobilis and mixtures thereof.

6. Tablet according to any one of claims 1-5, wherein said lactic acid bacteria is selected from the group consisting of the following strains: Lactobacillus rhamnosus LRH 28 (deposited at Belgian Coordinated Collections of Microorganism Nr. LMG 27341 ), Bifidobacterium breve Bbr 8 (deposited at Belgian Coordinated Collections of Microorganism Nr. LMG P-17501 ), Lactobacillus plantarum 14D (Coleccion espanola de cultivos tipo nr. CECT 4528), Streptococcus thermophilus

Z57 (deposited at Belgian Coordinated Collections of Microorganism Nr. LMG P-21908) and mixtures thereof.

7. Tablet according to any one of claims 1-6, comprising the following mixture of lactic acid bacteria:

Lactobacillus rhamnosus LRH 28 9% (w/w)

Bifidobacterium breve Bbr 8 50% (w/w)

Lactobacillus plantarum 14D 18% (w/w)

Streptococcus thermophilus Z57 18% (w/w).

8. Tablet according to any one of Claims 1-7, which is chewable.

9. A method for the preparation of the tablet of anyone of claims 1-8, characterized in that the compressing step is of the cold direct type, in particular at very low compression.

10. Tablet according to any one of Claims 1 to 8, packaged in a container comprising a moisture absorber.