Nutraceutical or pharmaceutical composition comprising bromelain

Abstract

The present invention relates to a composition of substances, preferably obtained from natural sources, that is effective in the prevention and / or treatment of inflammatory diseases. The composition of the invention comprises bromelain, phosphatidyl serine, phosphatidylcholine and starch. The composition may optionally comprise hydroxypropylcellulose. This formulation gives the composition gastroresistance properties, increased permeability across membranes, controlled release and high bioavailabil ity of the bromelain active ingredient. The composition of the invention is prepared in solid, semi-solid or liquid pharmaceutical dosage form, preferably for oral administration.

Description

[0001] Nutraceutical or pharmaceutical composition comprising bromelain

[0002] DESCRIPTION

[0003] The present invention relates to a nutraceutical or phar- maceutical composition that is effective in the prevention and / or treatment of inflammatory diseases in both humans and animals.

[0004] Background of the invention

[0005] PHLOGOSIS

[0006] Phlogosis, also known as inflammation, is the set of chang- es and variations that occur in a specific part or in a larger area of the body as a result of a damage. It is an adaptive-nature response usually triggered by harmful stim- uli or conditions. In the genesis of pro-inflammatory con- ditions, physical agents such as trauma or heat, chemical agents such as acidic substances or biological agents such as bacteria and viruses may come into play. Thus, it can be understood that it is an important defence mechanism in the event of tissue damage or during infections, acting in the interest of restoring homeostasis. Inflammation, in general terms, is mainly an acute and local reaction, but can in various cases become chronic and affect increasingly larger areas of the body.

[0007] There are five key points of phlogosis and they are defined as calor, an increase in temperature of the affected area caused by increased vascularisation, tumor, swelling of the affected area due to the formation of exudate following vasodilatation, rubor, reddening following hyperemia, dol- or, a painful sensation triggered by the stimulation of the sensory endings by the triggering agent of the inflammation or the components of the inflammatory response, and final- ly, functio laesa, which consists in the functional impair- ment of the affected part.

[0008] Inflammation can be induced either by endogenous factors or exogenous factors.

[0009] In the case of exogenous factors, these can be further di- vided into two groups, those of a microbial nature and those of a non-microbial nature. The factors of a microbial nature that induce inflammation are PAMPs (pathogen- associated molecular profiles) and virulence factors, while those of a non-microbial nature include allergens, irri- tants, foreign bodies and toxic compounds.

[0010] Instead, endogenous inducers of inflammation are signals produced by stressed, damaged cells or tissues with altered function.

[0011] These inducers of inflammation activate the production of numerous mediators that act mainly on vascularisation and leucocyte recall. Some mediators such as histamine and ser- otonin are pre-formed and stored within mast cells, baso- phils and platelets, ready for use when needed, while oth- ers are synthesised ex-novo. The mediators of inflammation are subdivided into 7 groups based on their biochemical properties: vaso-active amines, vaso-active peptides, lipid mediators, fragments belonging to the complement system, cytokines, chemokines and proteolytic enzymes.

[0012] Vaso-active substances play a complex and important role in the vascularisation by causing vasodilation and increased vascular permeability. Lipid mediators derive from phospho- lipids of the cell membrane and have, as their main compo- nent, arachidonic acid, which can be metabolised by cyclo- oxygenases (COX-1 and COX-2) and become prostaglandins act- ing as an important algesic agent and inducer of fever or be metabolised by lipoxygenases and become leukotrienes and lipoxins .

[0013] Fragments belonging to the complement system act by promot- ing the recruitment of granulocytes and monocytes and in- duce mast cell degranulation. Pro-inflammatory cytokines such as TNF-α, IL-6, IL-1 and many others are produced by various cell types, in particular mast cells and macrophag- es, and are important in the inflammatory response as they are involved in the activation of endothelium and leuko- cytes. Chemokines, on the other hand, control leukocyte ex- travasation and chemotaxis. Finally, proteolytic enzymes have different roles ranging from body defence to tissue remodelling .

[0014] All these components contribute to the elimination of harm- ful stimuli and help restore the physiological conditions of the tissue.

[0015] However, under certain conditions, the inflammatory re- sponse can get out of control and it can turn, from a de- fence mechanism aimed at restoring homeostasis, into a cause triggering or even exacerbating a disease.

[0016] The mechanism underlying inflammation, let alone all the components involved in it, is not yet perfectly known, but great strides have been made, especially in controlling the inflammatory response.

[0017] To date, a wide variety of anti-inflammatory agents are available on the market, agents that have demonstrated to be highly effective, however their use is not free from side effects.

[0018] The best known and most widely used anti-inflammatory drugs are the so-called NSAIDs (non-steroidal anti-inflammatory drugs) which act by inhibiting the two isoforms of the COX enzyme (COX-1 and COX-2). This mechanism prevents the for- mation of prostaglandins and thromboxane that are important pro-inflammatory factors. However, despite their broad and proven efficacy, prolonged use of this class of drugs is accompanied by non-negligible side effects, the most fre- quent of which are at gastro-intestinal and cardiovascular level.

[0019] Another class of drugs used in the treatment of inflamma- tion is SAIDs (steroidal anti-inflammatory drugs), also known as glucocorticoids. Anti-inflammatory activity is me- diated through various mechanisms of action, but the major mechanism of action is thought to be cytokine-induced inhi- bition of gene transcription. SAIDs have the ability to en- ter the cell and bind the cytoplasmic steroid receptor, subsequently this complex migrates into the nucleus and recognises and binds specific sequences on the DNA. This prevents the transcription factors NF-kB and AP-1 from act- ing.

[0020] Although glucocorticoids are widely used in the treatment of chronic inflammation- and autoimmune-based diseases, their use is associated with annoying and in some cases se- rious side effects such as impaired carbohydrate metabo- lism, osteoporosis, menstrual abnormalities, hyperlipidae- mia, decreased GH secretion, and they also have important interactions with other drug classes. Side effects are usu- ally directly proportional to the duration of the treatment and the dosage used.

[0021] New drugs, called monoclonal antibodies or more simply bio- logical antibodies, have the ability to act and inhibit the various activities of various pro-inflammatory agents in a highly selective manner. However, even these are not free from side effects; indeed, the greatest side effect is a reduction in the body's defences against other harmful stimuli and infections.

[0022] Inflammation thus intervenes in a large number of diseases that can affect all parts of the body. Inflammatory diseases at the osteo-articular, uro-genital, dental and gynaecological levels in both humans and animals are of particular interest for this Patent Application.

[0023] In relation to osteo-articular inflammatory diseases, ar- thritis and osteo-arthritis may be commonly mentioned.

[0024] The uro-genital tract is also often subject to inflammatory states, especially with advancing age. The pro-inflammatory state is frequently associated with infections cause by bacteria and viruses, but also with a wide range of non- infectious processes. The most common inflammatory process- es are mastitis, vulvovaginitis, metritis, endometritis, prostatitis, epididymitis, orchitis. All pathologies that, unless treated properly and correctly, can lead to infer- tility.

[0025] In dentistry, inflammation plays an important role in vari- ous areas, both in the case of infections, and in case of systemic diseases, or damage caused by a physical agent. Phlogosis is at the root of stomatitis, which is a wide- spread inflammation of the oral cavity, but periodontitis, gingivitis and aphthosis can also occur. Furthermore, the role of phlogosis in case of tooth extraction should not be underestimated, as pain and swelling are important limiting factors in this procedure.

[0026] GLOSSARY

[0027] The terms used in this description are as generally under- stood by the person skilled in the art, unless otherwise indicated.

[0028] The term "extract", in the context of the present descrip- tion, refers to any product attributable to a herbal drug including all products derived from mechanical processing (pulverisation, crushing, mixing and / or other methods) or extractive treatments (solvent extraction, distillation, and / or other specific methods) performed on a drug.

[0029] DETAILED DESCRIPTION OF THE INVENTION

[0030] Bromelain is the generic name for a family of proteolytic enzymes containing the sulphydryl group. But the beneficial effects of bromelain can also be attributed to non- proteolytic constituents such as peroxidases, acid phospha- tases, various protein inhibitors and organic calcium.

[0031] Bromelain can be synthetically or naturally derived; the main natural source of bromelain is the extract of Ananas comosus.

[0032] Different units of measurement can be used to assess the enzymatic activity of bromelain, but the one most commonly used is gelatin dissolving units (g.d.u.).

[0033] By virtue of its historical and widespread use, several tests have been carried out both in vitro and in vivo, both to assess its pharmacokinetics, but above all to assess the mechanism of action through which it is able to act and play such an important role in the treatment of various diseases.

[0034] To date, the mechanism of action is not yet fully elucidat- ed, but several pathways and mediators are indeed involved. Its anti-inflammatory activity is mainly attributable to its ability to inhibit bradykinin formation at the site of inflammation by acting on the kinin-kallikrein system, as well as its ability to limit fibrin production, thereby re- ducing intermediaries in the coagulation cascade.

[0035] In vivo studies demonstrate that bromelain in a mouse model of carrageenan-induced inflammation had the ability to sig- nificantly reduce the production of prostaglandins and sub- stance P, two important mediators of the inflammatory and pain response. Bromelain was already effective at a dose of 10 mg / kg. In addition, the activity of bromelain in vitro was assessed in the aforesaid study, where, however, it had no effect; on the contrary, the substance P value in- creased. This demonstrates that bromelain does not act di- rectly on these mediators, but implicitly, resulting in a decrease of the two inflammatory markers.

[0036] Furthermore, bromelain appears to have the ability to in- tervene and influence different cell signalling cascades, acting as signalling molecules, many of which are involved in cell survival.

[0037] In a carcinogenesis mouse model it was demonstrated that treatment with 1 mg / animal of bromelain caused an increase in the regulation of p53 and Bax, two important transcrip- tion factors involved in the process of apoptosis, and sub- sequent activation of caspase 3 and caspase 9 with concomi- tant decrease in Bcl-2.

[0038] In addition, a marked inhibition of the expression of COX-2 and the transcription factor NF-kB was obtained, by virtue of the fact that bromelain showed the ability to block the phosphorylation and subsequent degradation of IkBα, which is also an important factor inhibiting NF-kB activity. This results in marked anti-inflammatory and carcinogenesis- preventive activity.

[0039] In addition, bromelain at a dose of 5 mg / ml demonstrated the ability to decrease platelet activity and aggregation in vitro in blood samples taken from 10 healthy, non- smoking male volunteers (Glaser et al.).

[0040] In addition, bromelain treatment in vitro showed the abil- ity to decrease platelet adhesion to endothelial cells, and probably its activity as an anti-platelet agent is mainly related to this ability.

[0041] The anti-thrombotic capacity of bromelain when administered orally at a dose of 60 mg / kg was evaluated in a mouse model of laser-induced thrombosis. Under these conditions, brome- lain demonstrated the ability to significantly inhibit thrombus formation in mesenteric vessels in rats.

[0042] All these scientific evidence demonstrated in vitro and in vivo, was subsequently also highlighted in clinical stud- ies. Bromelain has proven effective in the treatment of in- flammation-based diseases.

[0043] In the trial carried out on 84 patients who had a surgical removal of the mandibular third molar, the administration of 40 mg bromelain every 6 hours for 6 days in combination with an injection of 4 mg dexamethasone proved effective in significantly reducing the intake of anti-inflammatory drugs and in addition, reduced post-operative swelling, although not significantly.

[0044] Bromelain has proved to be a valuable ingredient in post- operative inflammatory conditions in clinical trials; one study demonstrated an effective improvement in swelling, pain and difficulty in swallowing in the groups treated with various dosages of bromelain compared to the placebo group.

[0045] Despite the important properties discussed, the proteolytic activity of bromelain is rapidly inactivated due to its in- stability at acidic stomach pH and / or denaturation, aggre- gation phenomena that it undergoes after oral administra- tion that may result in low therapeutic efficacy.

[0046] For this reason, the object of the present invention is a technologically innovative formulation containing bromelain in combination with phosphatidylserine, phosphatidylcholine and starch, in particular modified starch, capable of en- hancing the stability of bromelain in the gastric environ- ment, improving absorption and bioavailability thereof, and thus optimising its therapeutic effect. This formulation which is the object of the present inven- tion ensures gastro-protection and controlled release of bromelain through the combination with the three functional excipients such as phosphatidylcholine, phosphatidylserine and starch.

[0047] In particular, starch, preferably acetylated modified starch protects the phospholipid-bromelain combination from the gastric environment and allows it to arrive in high concentrations in the intestine where, thanks to the pres- ence of phosphatidylcholine and phosphatidylserine, brome- lain is optimally absorbed.

[0048] The object of the present invention is therefore to make available a nutraceutical or pharmaceutical composi- tion adapted to deliver bromelain to improve efficacy, safety and compliance in patients with inflammatory diseas- es.

[0049] A pharmaceutical product or a food supplement comprising the nutraceutical or pharmaceutical composition according to the invention also fall within the scope of the invention. In addition to the active ingredient, bromelain, and the func- tional excipients starch, phosphatidylcholine and phosphati- dylserine, the pharmaceutical product or food supplement of the invention may optionally comprise further active ingredi- ents and functional excipients, which are easily chosen by the person skilled in the art according to the needs. The choice of vehicles, excipients and / or diluents required for the for- mulation of the pharmaceutical product or food supplement into an appropriate dosage form also falls within the normal abili- ties of the person skilled in the art.

[0050] The present invention may also optionally contain hydroxypro- pylcellulose.

[0051] Hydroxypropylcellulose is a cold water-soluble cellulose de- rivative.

[0052] It is widely used in the pharmaceutical industry as a binder and is a recognised safe food additive (E463).

[0053] The nutraceutical or pharmaceutical composition accord- ing to the invention is as defined in the appended claim 1.

[0054] Further features and advantages of the invention are de- fined in the dependent claims. The claims also form an inte- gral part of the present description.

[0055] A detailed description of some preferred embodiments of the invention is given below.

[0056] As indicated, the nutraceutical or pharmaceutical com- position of the present invention comprises bromelain as the active ingredient and a combination of functional ex- cipients that provide the composition with gastro- resistance properties, increased resistance to degradation, controlled release and increased bioavailability. This com- bination of functional excipients includes phosphatidylcho- line, phosphatidylserine and starch.

[0057] The nutraceutical or pharmaceutical composition of the invention is therefore particularly effective for the de- livery and supplementation of bromelain for the prevention and / or treatment of inflammatory diseases. Examples of such diseases are gingivitis, periodontitis, stomatitis, inflam- mation and pain from tooth extraction, vulvovaginitis, me- tritis, endometritis, prostatitis, epididymitis, orchitis, mastitis, osteoarthritis, arthritis, soft tissue oedema.

[0058] Starch is an organic compound of polysaccharide nature consisting of the repetitions of glucose units linked by a- glycosidic bonds. It consists of two types of polymers: am- ylose, which generally amounts to about 20%, and amylopec- tin, which generally amounts to about 80%. Amylose forms the central part of starch granules, is soluble in very hot water and consists of glucose molecules linked by a-1,4 glycosidic bonds. Amylopectin is a polymer with a high de- gree of branching that forms the outer part of the gran- ules. The monomeric units composing it are joined at the branching points by a-1,6 glycosidic bonds. In nature, it is formed in the green parts of plants and is then accumu- lated in reserve organs such as tubers, seeds and roots. Due to its properties and characteristics, it has always been industrially used.

[0059] Starch is particularly important in the food industry, which uses it as a thickening agent and in the production of sweeteners such as maltitol and sorbitol. Due to its ad- hesive properties, it is also used in the production of pa- per and glues, in the form of starch solution. In the phar- maceutical industry, starch has always been used as an ex- cipient and to form coatings due to its binding properties.

[0060] Although starch is also used in its natural form, com- panies are mainly interested in modified starches, i.e. starch molecules suitably modified to meet the needs of the various production processes in which starch is used. Such modified starches can be obtained by using, as a source, plants that have undergone natural or induced genetic muta- tions and which thus produce starches with altered charac- teristics. Another strategy is to modify starch, generally derived from maize, tapioca and rice, by means of chemical (addition of functional groups, treatment with acids and bases), physical (gelatinisation) or enzymatic (partial hy- drolysis) treatments. Dextrins are an example of modified starches obtained by hydrolysis and re-polymerisation. These reactions can be carried out either by simple thermal degradation or by acid catalysis. The result is obtaining molecules characterised by shorter chains and thus partial- ly or totally soluble in water. Examples of dextrins are cyclodextrins and maltodextrins, excipients widely used in the nutraceutical and pharmaceutical field.

[0061] The nutraceutical and pharmaceutical industry has shown great interest in modified high amylose starches (HASs). For a starch to be defined as such, it must have an amylose percentage of at least 50%. Starch with a high amylose con- tent can be obtained from genetically modified plants or by enriching with amylose starches that possess low percent- ages thereof. The strategies employed by several companies provide to use HASs for the preparation of solid pharmaceu- tical forms or in coating processes. HASs have several ad- vantages over other types of starch, such as better con- sistency, higher thermal stability, and greater resistance to moisture and adhesion phenomena. Several strategies have been put in place to exploit the advantages of HASs in the most appropriate manner. The Scherer Corporation, for exam- ple, is credited with the use of soft gel capsules in which a certain percentage of gelatine is replaced with the aforesaid starch. The capsules thus obtained look better and are more durable. The Dow Chemical Company can boast the use of more uniform capsules with greater stability in water and at high temperatures due to the use of hydroxy- alkylated HAS. The Upjohn Company boasts the use of amylose acetate phthalate as a coating agent in gastro-resistant preparations .

[0062] Different types of starch can be used in the present invention. By way of example, the following are mentioned: chemically unmodified pregelatinised maize starch, or acet- ylated pregelatinised maize starch with a high amylose con- tent, which, in this specific case, can be of up to 90% by weight. The percentage of acetyl groups is between 0.5% and 2.5%.

[0063] Acetylation is carried out, for example, with acetic anhydride, which guarantees a percentage of acetyl groups greater than 0.5% but not exceeding 2.5%. The starch prege- latinisation treatment consists in dispersing the acetylat- ed starch in water and subjecting the resulting dispersion to temperatures ranging from 100 to 130 degrees and high pressure. The starch granules subjected to this procedure burst and form a gel with a moisture content ranging from 5 to 10%. Once solidified and removed, the modified starch thus obtained can be used in processes for coating hard, soft and microgranule capsules and ensures that a coating is obtained that is both resistant and adequately viscous at the same time, that is able to mask unpleasant smells and flavours, and that can also be used if a gastro- resistant or modified-release pharmaceutical form is de- sired. The characteristics of the pharmaceutical form can be modulated by modifying the amount of starch used in the coating.

[0064] Tests carried out using the aforesaid starch have shown the gastro-resistant, moisture-protective action and abil- ity to release the active ingredient after a few minutes in the intestinal environment.

[0065] As indicated, further functional excipients present in the nutraceutical or pharmaceutical composition of the in- vention are phosphatidylcholine and phosphatidylserine. These compounds are part of the wide class of phospholip- ids. Phospholipids are molecules that are structurally very similar to triglycerides. They are composed of a glycerol molecule esterified at position 1 and 2 with fatty acids. The fatty acids that enter into the composition of natural phospholipids can be between 12 and 22 carbon atoms long; in position 1, a saturated fatty acid is generally present; in position 2, unsaturated fatty acids are found; in posi- tion 3, a phosphate group is present, which, in turn, is esterified with a complex molecule of various kinds such as choline, serine, ethanolamine or inositol. These molecules are the ones that give the phospholipid its name (phospha- tidylcholine, phosphatidylserine, phosphatidylethanolamine and phosphatidylinositol) and influence the physical prop- erties thereof as they determine whether the molecule is anionic (as phosphatidylserine) or zwitterionic (as phos- phatidylcholine) . The peculiar chemical structure makes phospholipids amphiphilic molecules, i.e. molecules capable of interacting with both polar and apolar solvents. More precisely, the fatty acid carbon chains represent the apo- lar portion that interacts with apolar solvents, while the phospholipid head, consisting of the phosphate group and the molecule bound thereto, represents the polar portion that interacts with water and other polar solvents. This structural characteristic makes them molecules with surfac- tant properties that above a certain temperature, defined as the critical micellar concentration, aggregate to form characteristic complexes that can vary in shape and size depending on the conditions of the environment wherein they are formed and the length of the fatty acid chains that make them up. For example, when dispersed in an aqueous so- lution, they form micelles with a typical shape, with the polar heads pointing outwards towards the aqueous environ- ment and the hydrophobic tails pointing inwards. When dis- persed in organic solvents, they form so-called inverse mi- celles, wherein the heads are turned inwards and the tails towards the apolar external environment. In terms of size, they vary according to the length of the carbon chains of the fatty acids that make up the phospholipids.

[0066] Phospholipids are molecules with very important biolog- ical significance, firstly because they take part in the composition of biological membranes, and secondly because they are involved in several complex mechanisms such as transduction of intracellular signals, regulation of the intracellular concentration of certain ions and mediation of the inflammatory processes as sources of arachidonic ac- id. Phospholipids are also interesting molecules from a technological point of view and are therefore used in the pharmaceutical and nutraceutical fields as technological aids in the formulation of delivery systems for various ac- tive ingredients. Many active ingredients may in fact have poor bioavailability due to the difficulty in crossing the barriers and biological membranes of many parts of the body. Phospholipids, and in particular phosphatidylcholine and phosphatidylserine, are valuable aids in the technolog- ical strategies for releasing various active ingredients. One of the advantages of phospholipid-based delivery sys- tems is the compatibility of phospholipids with cell mem- branes, both at mucosal and skin level. Phosphatidylcholine and phosphatidylserine, therefore, act as intestinal and topical absorption enhancers and this action can be as- cribed to the following mechanisms of action: thanks to their properties and structure, they can fuse with the lipids of the stratum corneum and membranes and alter the structure thereof, allowing various substances to pass through; in contact with intestinal fluids, they form micelles that contribute to increase the absorption of the active ingredients of interest as they extract lipids from mem- branes and alter the rheological properties, fluidity and composition of membranes by increasing permeability; the aforesaid micelles protect the active ingredients from chemical and enzymatic degradation and can be absorbed in the enterohepatic circulation of bile salts, together with mixed micelles from the diet, and can be transported to the bloodstream where they release the incorporated ac- tive ingredients.

[0067] The present invention thus makes it possible to achieve:

[0068] Gastro-resistance - Increased permeability across biological membranes

[0069] - Controlled release of bromelain

[0070] - Increased bioavailability of bromelain

[0071] The present invention constitutes a prompt and useful intervention for the prevention and / or treatment of inflam- matory diseases. This effect is ascribed to the combined action of its constituent substances. The starch used in the present invention protects bromelain from stomach acid pH and enzymatic degradation, ensuring the controlled re- lease thereof in the intestine. Phosphatidylcholine and phosphatidylserine increase the bioavailability of brome- lain due to the multi-mechanism action in the intestinal mucosa.

[0072] The efficacy of the nutraceutical or pharmaceutical composition object of the present invention is assessed us- ing the experimental protocol described below.

[0073] In order to assess gastro-resistance, a disintegration test is performed, as prescribed by Pharmacopoeia. A sample of the pharmaceutical form to be tested is placed in an in- strument containing 0.1 N hydrochloric acid. The gastro- resistant pharmaceutical form, in contact with the buffer at pH 2 for two hours, does not disintegrate. Subsequently, the sample is transferred to a buffer at pH 6.8 wherein it disintegrates within ten minutes.

[0074] Gastro-resistance can also be assessed by means of the Pharmacopoeia dissolution test, according to which the pharmaceutical form is placed in contact with a 0.1 N solu- tion of hydrochloric acid and must release less than about 20% of bromelain after 2 hours in order to satisfy the as- say.

[0075] In order to assess intestinal permeability, an in vitro test is performed, for example, on Caco-2 cells. The cul- tured cells are prepared using an appropriate growth medium (e.g. containing FBS, foetal bovine serum) and kept under controlled conditions (e.g. 37°C, in an atmosphere of 5% CO2and 100% humidity). After several passages, the cells are washed and pre-incubated with PBS. Following treatment with the formulations of interest, the samples taken are submitted for analysis to quantify bromelain.

[0076] The efficacy of the present invention can be assessed by monitoring the anti-inflammatory and / or anti-edemigenous activity of bromelain formulated as per claim 1 of the pre- sent invention compared to bromelain as such using methods known to the person skilled in the art (e.g., assessment of inhibition of pro-inflammatory cytokines such as for exam- ple IL-6, TNF-a, assessment of stimulation of anti- inflammatory factors such as for example IL-10, assessment of proteolytic activity) in in vitro or in vivo models.

[0077] The nutraceutical or pharmaceutical composition of the present invention is particularly effective in the preven- tion and / or treatment of inflammatory diseases due to the synergistic action of its components. As mentioned above, the nutraceutical or pharmaceutical composition of the present invention is incorporated into a pharmaceutical product or food supplement, which is formu- lated in a suitable dosage form, whose composition and preparation falls within the abilities of the person skilled in the art.

[0078] In a preferred embodiment, bromelain in the nutraceuti- cal or pharmaceutical composition of the invention is pre- sent in an amount ranging from 0.1 to 90%, preferably rang- ing from 0.5 to 80%, even more preferably ranging from 1 to 70%, with respect to the total weight of the composition object of the present invention.

[0079] By way of example, further percentages of bromelain that may be used in the composition of the invention are: 2%, 3%, 4%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 75%, or 85%.

[0080] In another preferred embodiment, the starch in the nutraceutical or pharmaceutical composition of the inven- tion is present in an amount preferably ranging from 0.1 to 90%, more preferably ranging from 0.5 to 80%, even more preferably ranging from 1 to 70% with respect to the total weight of the composition object of the present invention.

[0081] By way of example, further percentages of starch that may be used in the composition of the invention are: 2%, 3%, 4%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 75%, or 85%. In still another preferred embodiment, the phosphati- dylcholine in the nutraceutical or pharmaceutical composi- tion of the invention is present in an amount ranging from 0.01 to 50%, preferably ranging from 0.05 to 30%, even more preferably ranging from 0.1 to 10% with respect to the to- tal weight of the composition object of the present inven- tion.

[0082] By way of example, further percentages of phosphatidyl- choline that may be used in the composition of the inven- tion are: 0.02%, 0.03%, 0.04%, 0.06%, 0.07%, 0.08%, 0.09%, 0.2%, 0.3%, 0.4%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 15%, 20%, 25%, 35%, 40%, or 45%.

[0083] In still another preferred embodiment, phosphatidylser- ine in the nutraceutical or pharmaceutical composition of the invention is present in an amount ranging from 0.01 to 50%, preferably ranging from 0.05 to 30%, even more prefer- ably ranging from 0.1 to 10% with respect to the total weight of the composition object of the present invention.

[0084] By way of example, further percentages of phosphatidyl- serine that may be used in the composition of the invention are: 0.02%, 0.03%, 0.04%, 0.06%, 0.07%, 0.08%, 0.09%, 0.2%, 0.3%, 0.4%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 15%, 20%, 25%, 35%, 40%, or 45%.

[0085] All the above preferred embodiments may be combined with each other.

[0086] The pharmaceutical product or food supplement, which comprises the pharmaceutical or nutraceutical composition of the invention, is formulated in a preferably oral phar- maceutical form, which may be solid, semi-solid or liquid.

[0087] By way of example, a powder, an orosoluble powder, a granulate, a hard capsule, a soft-gel capsule, a tablet, a sachet, a solution, a syrup, a suspension or an emulsion are mentioned.

[0088] Non-limiting examples of nutraceutical or pharmaceuti- cal compositions object of the present invention are given below. As indicated above, such nutraceutical or pharmaceu- tical compositions are formulated as pharmaceuticals or food supplements and are administered in a suitable oral dosage form, possibly divided into one or more dosage units, such as a capsule, a tablet or a sachet.

[0089] The following examples are provided for illustrative purposes only and are not limiting the scope of the inven- tion as defined by the appended claims.

[0090] EXAMPLES

[0091] EXAMPLE 1 EXAMPLE 2

[0092] EXAMPLE 3

[0093] EXAMPLE 4

[0094] EXAMPLE 5

[0095]

[0096] EXAMPLE 6

[0097] In order to prepare the product "Bromelain Granulate" the fluidised bed granulation technology may be used. An example of preparation is given below, applied to the com- positions of the Examples.

[0098] The manufacturing process consists of the following steps: a) Mixing:

[0099] The raw materials, previously loaded into the basket of the granulator, are subjected to an initial mixing step on a fluid bed, with process air having a certain temperature (e.g. 80-90°C), until a mixture with an average temperature of approx. 44°C is obtained. During this step, a homogene- ous bulk is created in terms of composition and tempera- ture, which is a precondition for the optimal course of the subsequent granulation step. b) Granulation

[0100] The granulation step involves the addition of an aque- ous solution of a suitably chosen binding or granulating agent by direct spraying onto the bulk pre-mixed and fluid- ised on a fluidised bed. In this step also, process air is used, e.g. at 90°C, properly choosing the feed rate of the binding solution in order to obtain a granulate structured according to expectations (grain size, bulk density, flowability) and homogeneous. c) Drying

[0101] During the drying step, the water content of the pre- formed granulate tends to be restored to the conditions of the starting raw material mixture. The process air tempera- ture of the granulate at the end of the step is appropri- ately assessed in pilot tests according to this objective. d) Calibration

[0102] The semi-finished product obtained from the previous step is transferred from the fluidised bed granulator to an oscillating granulator through a sieve to reduce the parti- cle size of the coarser granules and agglomerates.

Claims

CLAIMS1. Nutraceutical or pharmaceutical composition comprising the combination of bromelain, phosphatidylcholine, phospha- tidylserine and starch.

2. Nutraceutical or pharmaceutical composition according to claim 1 comprising Hydroxypropyl cellulose.

3. Nutraceutical or pharmaceutical composition according to claim 1 and 2, for the use in the treatment or the preven- tion of inflammatory diseases in both humans and animals.

4. Nutraceutical or pharmaceutical composition according to claims 1 and 3 in wherein the bromelain is of synthetic origin and / or derived from an Ananas comosus extract.

5. Nutraceutical or pharmaceutical composition for the use according to claims 1 to 4, comprising a bromelain amount ranging from 0.1 to 90 %, preferably ranging from 0.5 to 80%, still more preferably ranging from 1 to 70% with re- spect to the total weight of the above mentioned combina- tion.

6. Nutraceutical or pharmaceutical composition for the use according to claims 1 to 5, comprising starch, preferably pregelatinized acetylated, preferably in an amount ranging from 0.1 to 90%, more preferably ranging from 0.5 to 80%, still more preferably ranging from 1 to 70% with respect to the total weight of the above mentioned combination.

7. Nutraceutical or pharmaceutical composition for the use according to any one of claims 1 to 6, comprising a phos-phatidylcholine amount ranging from 0.01 to 50%, preferably ranging from 0.05 to 30%, still more preferably ranging from 0.1 to 10% with respect to the total weight of the above mentioned combination and a phosphatidylserine amount ranging from 0.01 to 50%, preferably ranging from 0.05 to 30%, still more preferably ranging from 0.1 to 10% with re- spect to the total weight of the above mentioned combina- tion.

8. Nutraceutical or pharmaceutical composition for the use according to any one of claims 1 to 7, in which the inflam- matory diseases is chosen from the group consisting of gin- givitis, periodontitis, stomatitis, inflammation and pain from tooth extraction, vulvovaginitis, metritis, endometri- tis, prostatitis, epididymitis, orchitis, mastitis, osteo- arthritis, arthritis, soft tissue edema.

9. Pharmaceutical product or food supplement according to claims 1 to 8, formulated in a liquid, semi-solid or solid oral dosage form.

10. Pharmaceutical product or food supplement according to claim 9, in which the dosage form is a powder, an orosolu- ble powder, a granulate, a hard capsule, a soft-gel cap- sule, a tablet, a sachet, a solution, a syrup a suspension or an emulsion.

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