Patent Description:
In particular, the present invention relates to a topical, oral, pharmaceutical composition for use in the above-mentioned treatment, wherein said active ingredient consists of at least one probiotic (i.e. alive and viable) yeast capable of crossing the gastric barrier (i.e. gastric juices in the stomach and bile) undamaged and reaching the gut in alive and viable form.

More specifically, the present invention relates to a topical, oral, pharmaceutical composition for use in the above-mentioned treatment, wherein the active ingredient consists of at least the yeast Kluyveromyces marxianus fragilis B0399.

The diseases of the oral cavity that are of interest to the present invention are well-known inflammation-based diseases such as, for example, gingivitis, periodontitis, peri-implant mucositis and peri-implantitis.

Local inflammation is triggered by accumulation of gram-negative pathogenic bacteria (that are present in the plaque), which alter the composition of the normal oral microbiota (thus causing various forms of dysbiosis). The pathologic action of such bacteria is fostered and modulated, in different persons, by inflammation-promoting systemic factors. It can therefore be stated that local inflammation and systemic inflammation are bound by a bidirectional relationship. In this respect, it is worth reminding that, according to the scientific literature, all inflammatory forms involving oral tissues, and in particular all forms of periodontitis that lead to destruction of tooth supporting structures, allow for systemic absorption of gram-negative pathogenic bacteria, which therefore represent an important risk factor for numerous chronic systemic inflammatory diseases. Merely by way of example, these include arteriosclerosis with cerebrovascular accidents (doubled risk), coronary diseases (doubled risk), respiratory diseases (<NUM>- to <NUM>-fold risk), pregnancy complications (<NUM>-to <NUM>-fold risk), diabetes (<NUM>- to <NUM>-fold risk), and so on. The periodontal disease is a disease that cannot be healed, and the therapy that is traditionally used is mainly aimed at countering and limiting it.

The conventional therapy uses consolidated, tested and validated surgical and/or non-surgical hygienic-mechanic protocols. These are mainly based on the elimination of the triggering factor, i.e. the periodontal pathogenic gram-negative bacteria. The cornerstones of this therapy are, generally, good daily hygiene by the patient, associated with a non-surgical out-patient periodontal therapy at predefined intervals. In the most difficult and serious cases, a surgical periodontal therapy is also used, which is aimed at remodelling and/or rebuilding the anatomic components that have been seriously harmed by the disease.

Such conventional treatments also make use of antibiotics, disinfectants, antiseptics and/or antimicrobial agents such as, for example, chlorhexidine. However, such agents can only control, but not resolve, the disease, and may also have a number of side effects (anaphylactic reactions, transverse resistances, tooth pigmentation, and so on).

Recent approaches described in the scientific literature [e.g. <NUM>, <NUM>] and in the patent literature [e.g. <NUM>] propose the use of probiotics in the oral cavity.

For clarity's sake, it is necessary to remind beforehand that, when it comes to "probiotics", reference is made to microorganisms, bacteria or yeasts that are taken "alive and viable", so that they can reach the reference mucosa and "adhere thereto and multiply" to perform their different immune and metabolic functions over time. "Alive and viable" represents the fundamental characteristic inherent in a probiotic microorganism, which must be able to attach to mucosae and multiply thereon in order to replicate its functions over time. A dead yeast, or a part thereof, or a derivative thereof, since it cannot regenerate itself, can only perform a temporary administration-related activity.

It must also be said that "probiotic" is a definition that appears on circulars by the Italian Ministry, which in turn is the translation of a definition recognized at worldwide level ("microrganismi vivi e vitali che conferiscono benefici alla salute dell'ospite quando consumati in adequate quantità come parte di un alimento o di un integratore" ["alive and viable microorganisms that, when assumed in adequate amounts as part of a food or food supplement, confer a health benefit to the host"]: http://www. it/portale/temi/p2 <NUM>. jsp?id=<NUM>&ar ea=Alimenti%20particolari%20e%20integratori&menu=integrat ori). In brief, using probiotics (which are alive and viable) means providing the environment (organism or tissue) with functional elements capable of integrating their own functions according to requests from the environment itself, thus contributing to creating a symbiotic balance, the efficiency level of which defines the actual border between health and disease. These concepts have been recognized in the scientific literature for more than a hundred years, but are still unclear within the scope of a conventional medical approach to health problems.

Such definitions do not imply that such microorganisms are probiotics.

In addition, even assuming that such microorganisms are probiotics, they are "genetically modified" for the purpose of enhancing specific performance characters thereof, and this treatment, since it heavily modifies the genetic equipment of the microorganism, makes it different from the previous one.

For this reason, at this point, even assuming that it is still alive (which is uncertain and unlikely), it will be completely different from the initial microorganism. The document includes a long list of bacteria and yeasts that are modified in many ways and administered also by parenteral administration, intravenous injection, with antibiotics, in liquid carriers, but all of these conditions have nothing to share with a possible use of probiotics and give clearly to understand that these are not alive and viable microorganisms, but parts (peptides) of dead microorganisms.

<NUM> - <NPL>, explains how the alive yeasts taken into consideration can affect the immune system of man, and that such beneficial action is more efficient and complete when exerted by an alive microorganism rather than an extract obtained from (dead) parts of the same microorganisms.

Conventional treatments, as well as the latest ones mentioned above, suffer all from some drawbacks, such as, for example:.

As a consequence, a need is still felt for solving the technical problem of reducing the (local) inflammation of oral mucosae and tooth/implant supporting structures and also the (systemic) inflammation at gut level, simultaneously re-balancing the bacterial flora (microbiota) of both the mouth and the gut by means of the same active principle.

Therefore, industry operators still strongly demand for the availability of a pharmaceutical composition capable of topically treating the inflammatory states of oral mucosae and tooth/implant supporting structures and the diseases related thereto or derived therefrom, such as, for example, gingivitis, periodontitis, peri-implant mucositis and peri-implantitis, while also at the same time significantly reducing the inflammatory state of the gut, re-balancing the microbiota thereof, and thus ameliorating the systemic inflammatory state (overall health condition) of the individual.

It is the object of the present invention to provide an adequate solution to the above-described technical problem.

The present inventor has now found that a topical, oral, pharmaceutical composition in which the active ingredient consists of at least one suitable probiotic (eubiotic) yeast, i.e. alive and viable, capable of crossing the gastric barrier undamaged and reach the gut in alive and viable form, can provide an adequate (local/systemic) solution to the technical problem highlighted above.

It is therefore an object of the present invention to provide a topical, oral, pharmaceutical composition as defined above for use in the treatment of inflammation-based diseases of the oral cavity, as set out in the appended independent claim.

Some preferred embodiments of the present invention are set out in the appended dependent claims.

The preferred embodiments of the present invention illustrated in the following description only have an explanatory purpose and are by no means intended to limit the application scope of the present invention, which will be immediately apparent to those skilled in the art.

The present invention relates to a topical, oral, pharmaceutical composition for use in the treatment of inflammation-based diseases of the oral cavity, wherein said diseases are selected from the group consisting of gingivitis, periodontitis, peri-implant mucositis and peri-implantitis, and wherein the active ingredient consists of an effective amount of at least one probiotic yeast as defined in the claims, alive and viable, which can cross the gastric barrier undamaged and reach the gut in alive and viable form.

In said composition, said at least one probiotic yeast is present in an amount at least equal to <NUM> million cfu/dose (<NUM><NUM> cfu/dose); preferably, ≥ <NUM><NUM> cfu/dose; more preferably, ≥ <NUM>•<NUM><NUM> cfu/dose; even more preferably, ≥ <NUM>•<NUM><NUM> cfu/dose; even more preferably, ≥ <NUM><NUM> cfu/dose; even more preferably, ≥ <NUM><NUM> cfu/dose; for example, in the range of <NUM>•<NUM><NUM> cfu/dose to <NUM><NUM> cfu/dose.

In general, yeasts are a small part (<NUM>%) of the so-called oral and intestinal microbiota, while the remaining <NUM>% is represented by bacteria, mainly Lactic and Bifid ones. Yeasts contribute, e.g. when associated with Lactic bacteria, to exerting a general defense and support action (microbiota biodiversity). Therefore, yeast intake is generally important for keeping the microbiota in good efficiency conditions (territorial resilience). Merely by way of example, many articles in the literature magnify the positive effects of the yeast Saccaromyces boulardii both generally on the microbiota and specifically in the individual diseases involving the gut.

For the purposes of the present invention, the active ingredient consists of at the yeast is Kluyveromyces marxianus fragilis B0399.

This strain (produced, for example, by Laboratori Turval Italia, S. , Udine) was typified and filed at the BCCM-Belgian Coordinated Collections of Microorganisms, Culture Collection Mycoteque de l'Université Catholique de Lovain (Belgium) under code B0399 (April <NUM>, Access Number <NUM>). In <NUM> it was recognized as being probiotic (i.e. capable of ameliorating the balance of the microbial flora) by the Italian Ministry of Health.

This is the only probiotic yeast that is completely "natural" and "gut friendly", and is naturally found in food products (e.g. kefir from goat milk) and in the human intestine.

Kluyveromyces marxianus fragilis B0399 effectively performs many important functions, such as, for example: stimulating the growth and efficiency of an individual's beneficial endogenous bacterial flora (bifidobacteria); exerting an antimycotic action against Candida; fighting pathogens by lowering the pH; reinforcing the coating epithelia by adhering to the surfaces and taking nutrients away from them. Also of fundamental importance is its capability of producing short-chain fatty acids (SCFA), which are substances used by tissue cells in their energy production (Krebs) cycle to achieve better functional efficiency. Being active at low doses, B0399 crosses the gastric barrier undamaged, resists antibiotics, digests lactose and, most importantly, modulates the host's immune system by re-balancing the levels of cytokines and providing immunostimulating substances such as β-glucanes and oligosaccharides.

Such an active principle offers the operator several possibilities of use:.

When compared with lactic bacilli Lactobacillus ssp. , B0399 has properties and indications that imply greater efficiency:.

When compared with the yeast Saccaromyces boulardii, B0399 has properties and indications that imply greater efficiency:.

It is recommended by the producer (Turval) in cases of abdominal aerophagia, gut dismicrobism, IBS, dermatitis and Candida albicans.

To the inventor's knowledge, B0399, the beneficial effects of which at gut level are known, has until now been neither described nor used directly (i.e. topically) on mucosae and tissues of the oral cavity.

As already anticipated, the active ingredient of the topical, oral, pharmaceutical composition for use in the treatment of inflammation-based diseases of the oral cavity consists of an effective amount of at least the probiotic yeast Kluyveromyces marxianus fragilis B0399.

In said composition, said at least one probiotic yeast is present in an amount at least equal to <NUM> million CFUs per dose (<NUM><NUM> cfu/dose); preferably, ≥ <NUM><NUM> cfu/dose; more preferably, ≥ <NUM>•<NUM><NUM> cfu/dose; even more preferably, ≥ <NUM>•<NUM><NUM> cfu/dose; even more preferably, ≥ <NUM><NUM> cfu/dose; even more preferably, ≥ <NUM><NUM> cfu/dose; for example, in the range of <NUM>•<NUM><NUM> cfu/dose to <NUM><NUM> cfu/dose.

Expressing said amount in weight terms, said yeast may be generally present in the composition of the invention in an amount preferably ranging from a minimum of <NUM> to <NUM> per dose, more preferably from <NUM> to <NUM> per dose; more preferably from <NUM> to <NUM> per dose.

As far as the maximum dose is concerned, due to the absolute atoxicity of the microorganism and to the absence of any known unfavourable side effects, it can be stated that no maximum dose exists or may be conjectured; on the contrary, excessive dosage should reasonably do no harm, since it would ensure better efficiency. In such a case, the unused excess yeast would be eliminated by the body through the normal excretory ways.

In a preferred embodiment of the present invention, the microorganism (i.e. the at least one probiotic yeast of the invention) is provided in dehydrated form as powder obtained by known lyophilization processes and, depending on the different compositions and applications, can be administered in different carrier forms.

The dehydrated microorganism(s) of the invention has (have) the characteristic of reactivating when released in a suitable humid site, such as the oral mucosa or the intestinal mucosa, where colonization occurs to provide re-balancing (or eubiosis) among the various species of residential (autochthonous) or transient (allochthonous) bacterial flora.

In order to ensure appropriate crossing of the gastric barrier in all cases, the dehydrated microorganism(s) of the invention may optionally be microencapsulated to become resistant to gastric juices and/or bile. The optional microencapsulation is preferably effected through the use of well-known microencapsulation techniques commonly used in the industry, which will not be described in detail herein.

The pharmaceutical composition of the present invention for use in the treatment of diseases of the oral cavity as previously described may possibly also contain one or more additional active principles or substances known in the art, acting synergically, or performing an adjuvant or complementary function, with the alive microorganism(s) of the invention.

Said optional additional active principles of the present invention can be selected among many substances recognized by competent bodies as being miscible, e.g. probiotics, nutraceuticals, minerals, vitamins, "botanicals" vegetable preparations, prebiotics, enzymes, algae, and any other substances symbiotically compatible with the reference active principle (the yeast).

Merely by way of non-limiting example, said additional substances may be selected from:.

The compositions of the present invention may be for nutraceutical and/or pharmaceutical and/or cosmetic use, and preferably can be produced as a gel toothpaste, a gel for local surface applications, a miscible powder to be directly introduced into the periodontal pockets, or tablets, capsules, microcapsules or pearls, powders in packets or other containers, single-dose bottles, chewing gums, candies, pills or gels, solutions, emulsions in any form, sprays, mouthwash liquids, or any other technological form for oral administration commonly available in the industry.

Once activated in the mouth (as previously described), the principle can then be swallowed, so that it can continue its beneficial action also at gut level.

As a consequence, the compositions of the present invention may be swallowed after topical oral application, as opposed to being expelled by rinsing, since they are fully compatible with the human body. In particular, the characteristic of B0399 of being able to cross the gastric barrier undamaged allows the composition to exert a prolonged action on intestinal mucosae as well.

The following experimental section illustrates, merely by way of example and without by no means limiting the broad application potential of the present invention, some particularly preferred embodiments of the present invention. It goes without saying that all possible implementation and application variants of the same are within the grasp of a person skilled in the art, and as such they fall within the protection scope of the invention.

Using a conventional turbo emulsifier, the following substances are mixed, in this order and at room temperature: glycerine (<NUM> to <NUM>%), xanthan gum (<NUM> to <NUM>%), water (<NUM> to <NUM>%), silica (<NUM> to <NUM>%), lyophilized Kluyveromyces marxianus fragilis B0399 (<NUM> to <NUM>%), flavouring (<NUM> to <NUM>%), essential oil of lemon (<NUM> to <NUM>%), limonene (<NUM> to <NUM>%), citral (<NUM> to <NUM>%) in weight relative to the total weight of the composition. After the addition of each ingredient, the mixture is stirred for a time ranging from approx. <NUM> to <NUM> minutes until complete dispersion of the ingredient is attained. Once the toothpaste having the desired appearance/dispersion has been obtained, the product is collected into clean, sanitized drums and then packaged into sealed tubes.

The above toothpaste is applied onto the bristles of a toothbrush and interdental cleaning tools, and ensures a correct distribution of the active principle on all mucosae of the oral cavity. Laboratory tests have shown that <NUM> centimeter of toothpaste (recommended dose) to be spread over the bristles of a toothbrush contains an effective amount of the active principle taken into consideration herein. Daily use, possibly associated with a subsequent deglutition phase, ensures a natural beneficial (anti-inflammatory and curative) action on the involved tissues of the oral cavity and on the intestinal tract as well. Should the person decide to not swallow the product, it is however recommended to not rinse the mouth after use, so as to promote a prolonged local action of the active principle.

A gel, or a powder, containing the active principle (lyophilized Kluyveromyces marxianus fragilis B0399 in an amount ranging from <NUM> to <NUM> per <NUM> of composition), mixed with a suitable quantity of water or enriched with compatible or adjuvant substances, such as to maintain the nutraceutical formulation, such as typified bacteriocin-producing bacterial strains or phytotherapic active principles having beneficial effects that are adequate for the problem to be treated (rosa canina, acerola, aloe, elements having a prebiotic action, etc.), are introduced into suitable dental masks custom-made by the dentist and are applied onto the dental arches so as to keep the product in prolonged contact with the dental structures and the mucosae. This is a very effective solution for serious cases of periodontitis or peri-implantitis. In fact, the active principle, being protected by the mask, will remain for a long time highly concentrated and stably in contact with the structures to be treated.

As an alternative, said gel or said powder of Example <NUM>, possibly suitably enhanced and supplemented with other adjuvant additives appropriately selected among those previously described herein, may also be directly introduced into the periodontal pockets, e.g. by means of a syringe fitted with a thin needle/cannula, so as to exert a more immediate and direct action (this application must be made as an out-patient treatment by skilled and trained personnel).

A traditional chewing gum is prepared as commonly known in the art, wherein <NUM> of lyophilized Kluyveromyces marxianus fragilis B0399 per gum piece/dose and optionally an adequate amount of additional adjuvant substances (e.g. probiotics, prebiotics, etc.) are added to the mixture of desired ingredients. This type of solution turns out to be particularly effective when treating inflammatory diseases of the oral cavity. In fact, in addition to squeezing the active principle on teeth and gums, mastication also adds an effective mechanical cleansing action (plaque shattering), associated with recirculation of saliva, enriched with active principle, in the interdental spaces.

<NUM> chewable or swallowable tablets are prepared with methods and equipment commonly employed in the pharmaceutical art, adding <NUM> of lyophilized Kluyveromyces marxianus fragilis B0399 per tablet and, optionally, an adequate amount of additional adjuvant substances (e.g. probiotics, pre-biotics, etc.) to the mixture of desired ingredients. This type of solution may be the most appropriate one for persons who, for any reason, do not want to swallow toothpaste after brushing. Chewable or suckable tablets, similar to candies, will allow the active principle to remain in situ (i.e. in the mouth) while ensuring constant dosage and allowing the active principle to reach the gut in alive and viable form via saliva deglutition.

The compositions according to the present invention have advantageously proven to be useful in the dentistry field and, in general, in the medicine field for treating inflammatory diseases of the oral cavity; in particular, they have proven to be useful for treating diseases of the oral mucosa, gingivae, and tooth and/or implant supporting tissues.

Advantageously, the compositions according to the present invention have also proven to be useful in the therapeutic treatment of the dysbiosis of the intestinal microbiota.

Furthermore, the compositions have also turned out to be useful for treatments aimed at reducing halitosis caused by fermentation of protein-containing food residues, effected by proteolytic bacteria.

Claim 1:
A topical, oral, pharmaceutical composition for use in the treatment of inflammation-based diseases of the oral cavity, wherein the active ingredient consists of at least one probiotic yeast, alive and viable, capable of crossing the gastric barrier undamaged and reaching the gut in alive and viable form, said yeast being the Kluyveromyces marxianus fragilis B0399 and being present in an amount ≥ <NUM><NUM> cfu/dose.