WO2013160792A1

WO2013160792A1 - Compositions comprising ornithine alpha-ketoglutarate, processes for their preparation and their use.

Info

Publication number: WO2013160792A1
Application number: PCT/IB2013/052919
Authority: WO
Inventors: Paola Maffei; Annalisa Sforzini; Giuseppe Bottoni; Giuseppe Claudio Viscomi
Original assignee: Alfa Wasserman SpA
Current assignee: Alfa Wasserman SpA
Priority date: 2012-04-24
Filing date: 2013-04-12
Publication date: 2013-10-31
Anticipated expiration: 2014-10-24
Also published as: ITBO20120226A1

Description

COMPOSITIONS COMPRISING ORNITHINE ALPHA-KETOGLUTARATE, PROCESSES FOR THEIR PREPARATION AND THEIR USE.

Object of the invention

The present invention relates to compositions comprising ornithine alpha- ketoglutarate granules (OKG), characterized in that they have controlled release.

The invention also describes processes for their preparation and their use in the treatment and prevention of pathologies directly or indirectly related to states of malnutrition and/or malabsorption.

Field of the invention

Ornithine alpha-ketoglutarate (OKG) is a salt formed by two molecules of L(+) ornithine and one molecule of alpha-ketoglutarate. OKG stimulates the production of insulin and growth hormone and increases the synthesis of glutamine and other amino acids such as arginine and proline, as described by Cynober L. in Curr. Opin. Clin. Nutr. Metab. Care 2, 33-37 (1999).

Ornithine takes part in several metabolic processes. It is involved in the urea cycle, at whose end it is regenerated from arginine by releasing a molecule of urea and, after a transamination reaction, leads to the formation of glutamine- semialdehyde, which is a precursor of both glutamate and proline.

Alpha-ketoglutarate is involved in the Krebs cycle: it is a nitrogen acceptor, and therefore an amine acceptor, generating glutamine by means of transamination or amination reactions through glutamate-dehydrogenase in perivenous hepatocytes. The amidation of glutamate to glutamine by means of glutamine-synthetase takes place in several tissues and organs, such as muscles, lungs, liver and brain.

Ornithine and alpha-ketoglutarate are involved in different metabolic routes, but both of them share a common route leading to a glutamate increase. Several studies have showed that the OKG properties are due to the specific interaction of ornithine and alpha-ketoglutarate, which have a synergic effect.

Several clinical studies have showed the efficacy of OKG in patients who had been burned, were suffering from malnutrition or had been surgically treated. Moreover, OKG has anabolic properties in traumatic or stressful situations, improves bowel morphology and function, accelerates the healing process of wounds caused by burns or surgery, and stimulates the immune function. The OKG action mechanism, as described by Cynober L. et al. in "Ornithine alpha-ketoglutarate in Nutritional Support" Nutrition, Vol. 7 N. 5, 1991, is multi- factorial and related to the production of ornithine and/or alpha-ketoglutarate metabolites, acting on the protein metabolism and on the immune system, to the stimulation of the secretion of anabolic hormones such as insulin and growth hormone, and also to the improvement of the nitrogen balance.

Cynober et al., in Nutrition, Vol. 7 N. 5, 1991, also report the clinical study on patients undergoing surgery of the gastro-intestinal tract, wherein, after the OKG parenteral administration, it was observed an increase of insulin and growth hormone in blood, given by the stimulation of the anabolic hormones secretion. The same effect was also observed in patients suffering from cirrhosis and in healthy subjects.

The insulin stimulation is not related to the separate effect of ornithine or alpha-ketoglutarate, but the synergic effect of the two associated molecules and it is dose dependent.

It has been demonstrated that metabolites responsible for the insulin-secretion effect of OKG are, in particular, glutamine and nitrogen monoxide, arginine, whose glucagon secretion effect is well known, growth hormone (GH), prolactin and insulin- like growth factor 1 (IGF1). Cynober furthermore described the primary role of the main active OKG metabolites such as glutamine, arginine, proline and polyamines.

In particular, the effects shown by glutamine are such to allow the hypothesis that this is the main active OKG metabolite. Glutamine transports amine groups through cell membranes, thus being useful in ammonia detoxification; it is able to penetrate through the hematoencephalic barrier and entering the brain, wherein it is converted into glutamate, the most critical and widespread excitatory neurotransmitter of the central nervous system with immune and cerebral activity; it causes the volume increase of muscle cells by stimulating the protein synthesis; it plays a role in the over-tiring syndrome; it plays a role in the effort recovery; it stimulates the secretion of the growth hormone and has an anti-oxidizing action.

Arginine affects cell immunity, probably thanks to a mechanism related to the production of nitrogen monoxide.

In turn, proline seems to affect the healing properties attributed to OKG, since said amino acid is a precursor of hydroxy proline, contained by collagen.

Furthermore the increase of aliphatic polyamines intratissutal concentration in the bowel mucosa shows the beneficial action of OKG.

Several studies, like that described by Czernichow B. et al. in Gut 1997; 40: 67-72, confirmed the OKG trophic properties with regard to the bowel mucosa. Studies performed in animal models demonstrated that the enteral administration of OKG leads to a recovery of functions and bowel morphology damaged by events having different causes, thus observing an increase of the height of intestinal villi and a regeneration of the cells of intestinal crypts, favoring the renewed growth of the damaged mucosa. The OKG effect on the nitrogen balance can always be related to its amino acids metabolites. The nitrogen balance is negative in the case of burns, surgery, traumas, hypercatabolic state, since an increase of the protein balance leads to a clear protein loss at the expense of several organs and tissues, among which, in particular, skeletal muscles, which are a main source of amino acids for the organism. In these cases, some amino acids such as glutamine and arginine become indispensable, since their endogenous synthesis can no longer cover the body needs.

Cynober L. et al. in Nutrition, 2003, Vol. 19 (1), page 73-5, described the effect of OKG in elderly patients suffering from chronic malnutrition and malabsorption. It is well known that malnutrition affects a percentage higher than 40% of elderly patients, and that chronic malnutrition causes physiologic changes such as, for instance, loss of muscle mass, lack of protein synthesis and atrophy of digestive mucosa. Moreover, malnutrition is often related to other pathologies, such as, for instance, recurrent fractures and infections.

Therefore it is important to reach a balanced nutritional state in all subjects with problems related to malnutrition and, in particular, in elderly subjects, in order to avoid further complications which could worsen already existing pathologies and increase the physical wasting of the organism.

Some amino acids, and among them glutamine as well, are important regulators of protein synthesis, and can fight against functional and metabolic damage caused by a trauma.

Furthermore, glutamine plays a critical role in the metabolic adjustment after injuries, since it is a fuel for cell division and a precursor of glutathione, pyrimidines and purines, stimulates protein synthesis and therefore inhibits protein catabolism. It has been demonstrated that OKG increases appetite and body weight and accelerates healing processes in elderly subjects suffering from malnutrition. Brocker P. et al. in Age Ageing 1994 Jul, Vol. 23 (4), pages 303-6, described the beneficial effect of the OKG administration in patients recovering from serious illnesses, such as respiratory infections, and surgery. The effects were evaluated after 1 , 2 and 4 months of treatment, showing a remarkable improvement of nutritional state, appetite, a gain in weight and a general improvement of patients' s quality of life.

Therefore, OKG is useful in subjects suffering from related malnutrition and/or malabsorption states in patients undergoing surgery and traumas, and also in healthy subjects to increase their muscle mass and accelerate their recovery from physical stress.

Solid compositions have been studied with the main aim of avoiding OKG dissociation into its different components in the stomach in order to reach high OKG concentrations in the intestine.

EP 1806983B1 describes pharmaceutical compositions, nutritional and/or pharmaceutical preparation containing OKG, for the treatment of disturbance in micro elements absorption such as iron, zinc, copper, manganese, calcium, phosphorus, from the alimentary canal, in various diseases such as anemia, cardiac dysfunctions, atherosclerosis, osteoporosis, arthritis and bowel dysfunctions.

WO 2002/078676 describes pharmaceutical controlled release compositions containing amino acids linked to keto acids, in particular to OKG, and their use in subjects suffering from malnutrition and malabsorption problems. The described compositions have the form of controlled release granules, wherein an inert support is coated with layers containing OKG, and the obtained granules are coated with film comprising gastroresistant polymers. The controlled release granules described in WO 2002/078676 are obtained by means of a process comprising a first step wherein a solution containing OKG is sprayed on inert granules and a second step wherein the granules are coated with a gastroresistant polymer. The coating step is difficult and it is repeated thrice to obtain a complete coating, with a final yield of about 5%. The granules obtained by means of the aforesaid process contain an amount of OKG corresponding to about 0.06% by weight if compared to the granule weight; this means that in order to obtain an OKG amount comprised between 2 and 20 g, a subject should assume a total amount of granules comprised between about 3.4 kg and 17 kg.

Moreover, in order to obtain granules comprising an OKG amount comprised between 2 and 10 g, suitable for a single administration, the starting OKG amount should be larger than 20 kg, demonstrating that the process of WO 2002/078676 cannot be industrially employed for producing a product useful as a medicine or as a food supplement.

The production of solid compositions comprising OKG in controlled release granules is a complex process which must be developed taking into account the OKG high solubility in aqueous solution. The use of inert supports, as described in WO 2002/078676, does not solve the problem of obtaining controlled release OKG granules useful for the administration to humans or animals, since for obtaining a complete polymer coating of the granules the coating process must be repeated several times, with consequent lower yield in the final processes.

Therefore, there is a need for having available controlled release OKG granules containing OKG amounts larger than those of the known art, obtainable by means of a simple, reproducible and high- yield processes. Granulation is an important process, because it increases flowability, avoids segregation, increases density by decreasing volume, improves compressibility characteristics, reduces agglomeration phenomena and reduces the total amount of fine powders dispersed in the air.

Coating of particles with different and non-homogeneous sizes is complex, difficult and not reproducible. Two main techniques for obtaining granules are known in the art: dry granulation and wet granulation. Dry granulation does not involve the use of a wetting phase and involves a smaller risk of chemical and physical alterations of granulate materials. The powder is compacted by applying a mechanical force and the obtained solid mass is crushed and sieved until obtaining granules having the wished size. This kind of granulation is suitable for very bulky powders containing active principles which are scarcely stable under variable conditions of temperature and humidity.

Wet granulation involves a starting powder mixing step and then a "wetting" step, wherein the powder is wet and kneaded with a suitable amount of liquid phase.

The wet granulation processes known in the art are: kneading and granulation through nets; fluid bed granulation; granulation by extrusion - spheronization and granulation in high speed mixers-granulators (High-Shear Mixer).

The granules obtained by means of a wet granulation process must then be dried by means of drying system in static or dynamic mode, well known in the art.

Active ingredients soluble in aqueous solution, like OKG, make easy the granulation process, in particular the wet granulation process, but a careful development of the composition and parameter process in order to obtain granules having suitable sizes for the following coating steps has to be pointed out. The granule size and homogeneity degree are relevant parameters for the following coating steps, in particular for the coating with polymers suitable for controlled release. Granules having sizes smaller than 1 mm and high homogeneity degree are particularly requested in the production of pharmaceutical compositions. Granules having too large a size can give rise to agglomeration during their production process and clots formation, whereas granules of smaller size tend to be scattered in the air or to adhere to the walls of the production apparatuses and form lumps or agglomerates. Moreover granules with large size are not acceptable by patients and subjects.

The formation of granules with uneven distribution leads to finished products which are not reproducible, and product certification agencies for products for human or animal use, and in particular pharmaceutical products, require that the specifications on reproducibility and quality are strictly respected.

Pharmaceutical compositions containing not gastroresistant OKG are known and commercially available such as for instance Cetornan^®, for the treatment of malnutrition and malabsorption states.

OKG in composition not gastroresistant has the problem that in an acid environment like the stomach, dissociates itself into its two components, ornithine and alpha-ketoglutarate, therefore to obtain high concentrations of undissociated OKG in the intestine, high dosages of OKG should be assumed with the risk of possible side effects such as, for instance, diarrhea and meteorism.

Therefore, there is a need for available pharmaceutical and nutritional or alimentary controlled release compositions comprising OKG, containing an OKG amount larger than the one described in WO 2006/078676, obtained by means of simple and reproducible processes, scalable for high yield industrial production.

There is a further need for providing nutritional and/or pharmaceutical compositions which can be dispersed or suspended in drinks or semi-solid food, which are well accepted and tolerated by all subjects, in particular by patients suffering from related deglutition problems, for the treatment of chronic malnutrition and/or malabsorption states and by burned or surgically treated patients.

Summary of invention

The inventors of the present patent application have now identified new compositions comprising controlled release OKG granules, wherein said granules comprise an OKG amount from 30% to 80%> (w/w) in respect to the weight of the granules and they provide controlled release.

OKG compositions are scarcely soluble at pH lower than 5.0 and they are soluble at pH higher than 6.5.

OKG granules can be comprised in pharmaceutical or supplementary food compositions with acceptable excipients. The present application also describes processes for the preparation of controlled release OKG granules and composition comprising them.

The preparation of controlled release granules comprise a step of wet granulation in high shear granulator and the coating of granules with suitable polymers for providing controlled release.

The production yield with the described processes for obtaining controlled release ornithine granules comprising a step of wet granulation and coating is higher than 85%.

OKG controlled release granules are useful in the treatment and /or prevention of pathologies related to malnutrition or malabsorption states.

The acceptability, the tolerability and safety of oral sachets comprising OKG controlled release granules and Lactoferrin (OKG-LF) was assessed in a single centre, open trial on 42 elderly subjects. Detailed description of the invention

The present invention describes compositions comprising controlled release OKG granules wherein said granules comprise an OKG amount between 30% and 80% (w/w) with respect to the final weight of granules.

The described compositions comprise controlled release OKG granules, alone or together with pharmaceutically acceptable excipients, active principles and/or nutritional agents, such as, in an exemplificative but not limitative way, amino acids, vitamins, proteins, antioxidants.

The present invention also describes processes for obtaining controlled release granules and processes for obtaining pharmaceutical compositions comprising said granules.

The present invention also describes the use of the compositions comprising controlled release OKG granules for treating or preventing all pathologies related to alimentary malnutrition and/or malabsorption states with daily dosages comprised between 2.0 g and 20.0 g. It also describes the use of compositions comprising OKG granules as alimentary integrators, for all subjects who need them and for subjects suffering from physical fatigue.

The granules of the present invention comprise OKG in an amount comprised between 30%> (w/w) and 80%> (w/w), more preferably comprised between 45% (w/w) and 65%o (w/w), if compared to the weight of the finished granulate, together with acceptable excipients for human and/or animal use, and are coated with one or more polymers allowing a controlled release of OKG. In particular, the granules of the present invention comprise OKG together with one or more diluents, one or more binders, one or more plasticizers and one or more polymers suitable for controlled release.

OKG controlled release granules have the composition reported in Table 1.

Table 1

Compositions comprising controlled release granules of OKG, release OKG at pH values higher than 6.5, and release OKG in an amount smaller than 10% in a pH range comprised between 1.0 and 4.5.

A diluent suitable for the preparation of the controlled release granules object of the present invention is chosen in the group consisting of cellulose, microcrystalline cellulose, calcium phosphate, starch, kaolin, dehydrate calcium sulphate, calcium carbonate, lactose, saccharose, glucose, sorbitol, dextrose, mannitol, partially pregelatinized starch or their mixture.

A binder useful for the preparation of the controlled release granules is chosen in the group consisting of cellulose, cellulose derivatives, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose, starch, potato starch, maize starch, partially gelatinized starch, gums, synthetic gum, natural gum, polyvinylpyrrolidone, polyethylene glycol, gelatin, polyols, propylene glycol, alginates, sugars or their mixtures.

A plasticizer suitable for the preparation of the controlled release granules is chosen in the group consisting of glycerol, sorbitol, polyethylene glycols, triethyl citrate, triacetin or their mixtures.

The diluent is preferably chosen in the group consisting of cellulose, microcrystalline cellulose and mannitol. More preferably, the diluent is microcrystalline cellulose.

The binder is preferably chosen in the group consisting of cellulose, cellulose derivatives, starch, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, more preferably, the binder is hydroxypropyl methyl cellulose.

The plasticizer is preferably chosen in the group consisting of glycerol and polyethylene glycol, more preferably, the plasticizer is glycerol.

A polymer suitable useful for obtaining the OKG controlled release can be of synthetic or natural origin and they are chosen in the group consisting of acrylic acid copolymers, such as the methacrylic acid - ethyl acrylate copolymer 1 :1, methacrylic acid copolymers of acrylic or methacrylic ester such as the methacrylic acid - ethyl acrylate copolymer 1 :1, and the methacrylic acid - methyl methacrylate copolymer 1 :2, polyvinyl acetate phtalate, hydroxypropyl methylcellulose phtalate and cellulose acetate phtalate, products which are commercially available, for instance, with the trademarks Kollicoat^®, Eudragit^®, Aquateric^®, Aqoat^®; natural polymers such as shellac, commercially available with the trademark Aquagold^® (Shellac 25%) and ethyl cellulose. The polymer suitable for the preparation of OKG controlled release granules is selected among a natural polymer, such as shellac, commercialized in a 25% solution with the trademark Aquagold^®.

OKG granules can optionally have even bioadhesive properties to adhere to the bowel mucosa.

Examples of polymers, oligomers or their mixtures which can be included in the granules to give bioadhesive properties are chosen in the group comprising: pectins, zeins, casein, gelatin, albumin, collagen, kitosan, oligosaccharides and polysaccharides such as, for instance, cellulose, dextran, polysaccharides from tamarind seeds, xanthan gum, arabic gum, hyaluronic acid, alginic acid, sodium alginate.

When the bioadhesive polymer is a synthetic polymer, the polymer is chosen among polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl pyrrolidone, polysiloxanes, polyurethanes, polystyrenes, polymers of acrylic acid and methacrylic esters, copolymer of methacrylic acid-ethyl acrylate, polylactides, poly barbituric acids, polyanhydrides, polyorthoesters and their mixtures.

Further useful polymers are methyl cellulose, ethyl cellulose, hydroxy propyl cellulose, hydroxy butyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phtalate, carboxy methyl cellulose, cellulose triacetate, sodium salt of sulphate cellulose, polymethyl methacrylate, poly isobutyl acrylate, poly octadecyl acrylate, polypropylene, polyethylene glycol, polyethylene oxide, polyethylene terephthalate, polyvinyl acetate, polyvinyl chloride, polystyrene, polyvinyl pyrrolidone, polyvinyl phenol and their mixtures. Another group of polymers useful for obtaining bioadhesivity are polymers having a branching with at least a bound hydrophobic group, wherein the hydrophobic groups are generally non polar groups. Examples of such hydrophobic groups include alkyls, alkenyls and alkyl groups. In a preferred way, hydrophobic groups are chosen to increase polymeric bioadhesivity. Other polymers are characterized by hydrophobic branching with at least a hydrophilic group, such as carboxylic acid, sulphonic acid and phosphinic acid, neutral and positively charged amines, amides and imines, wherein the hydrophilic groups are able to increase the polymer bioadhesivity.

Controlled release OKG granules constituting a preferred object of the present invention are characterized in that they comprise an OKG amount comprised between

30.0% (w/w) and 80.0% (w/w) together with a diluent, such as microcrystalline cellulose, a binder, such as hydroxypropylcellulose, a plasticizer, such as glycerol, and a natural polymer suitable for obtaining its controlled release.

The preferred composition of the controlled release granules is reported in Table 2.

Table 2

Component Percentage (%) (w/w)

OKG 30.0 - 80.0

Micro crystalline cellulose 4.5 - 20.0

Hydroxypropyl methylcellulose 5.0 - 15.0

Glycerol 0.5 - 5.0

Shellac (Aquagold^®) 10.0 - 30.0 The controlled release OKG granules can be comprised in pharmaceutical or supplementary food compositions and can contain an OKG amount comprised between 0.5 g and 10.0 g. Preferably, the OKG amount comprised in the granules can range from 0.5 g to 8.0 g, more preferably from 0.5 to 5.0 g for a single dose.

The controlled release OKG granules compositions can be dispersed in water, aqueous drinks, fruit juices, milk, yogurt, semi-solid foods such as, e.g., puddings, soups, without limitations.

The compositions according to the present invention can comprise a variable amount of controlled release OKG granules useful for obtaining the wished dosage of a granule amount comprised between 10% (w/w) and 100%> (w/w) if compared to the weight of the finished composition. The compositions can optionally contain one or more active principles and/or one or more nutritional substances. Said substances can be comprised in an amount from 5% (w/w) to 70% (w/w) if compared to the weight of the finished composition.

The active ingredients and/or the nutritional substances possibly comprised in the OKG pharmaceutical composition are chosen among vitamins, in particular vitamin A, vitamin B complex, vitamin Bl, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B12, biotin, choline, folic acid, inositol, PABA, vitamin C, vitamin D, vitamin E, vitamin K; amino acids, in particular alanine, isoleucine, leucine, methionine, proline, tryptophan, valine, asparagine, cysteine, glutamine, serine, tyrosine, threonine, aspartate, glutamate, arginine, histidine, citrulline; proteins, in particular milk proteins such as, e.g., lactoglobulin, lactoferrin, lactoalbumin; antioxidants, polyphenols; microelements, such as phosphorus, chlorine, chromium, iodine, magnesium, manganese, molybdenum, potassium, selenium. According to a preferred embodiment of the present invention, the active principle or the nutritional substance comprised in the composition comprising controlled release OKG granules is chosen among milk proteins, such as e.g. lactoglobulin, lactoferrin, lactoalbumin. More preferably, the selected protein is lactoferrin.

The compositions for oral suspensions can be in the form of bags comprising OKG gastroresistant granules together with active ingredient and /or nutritional ingredient can comprise pharmaceutically acceptable excipients for instance suspending agents, which are useful for maintaining the granules in suspension in aqueous solutions.

The controlled release OKG granules mixed to a suspending agent can be comprised in the nutritional mixtures which can be administered by means of a nasogastric tube, a pharyngostomic tube, a gastrostomy tube, a digiunostomy tube.

The suspending agents suitable for the preparation of the composition according to the present invention can have natural or synthetic origin. The preferred suspending agents have natural origin, such as, e.g., agar-agar, gum tragacanth, guar gum, xanthan gum, carob seed flour, karaya gum, modified maize starch, carboxy methyl cellulose, microcrystalline cellulose or their mixtures in combination with other hydrocolloids. According to a preferred embodiment of the present invention, the suspending agent is formed by a mixture of xanthan gum, guar gum and carob seed flour, commercialized with the trademark Faramix^®.

A preferred composition in the form of granulate in bags for oral suspensions comprises an amount of controlled release OKG granules between 30.0% (w/w) and 95.0% (w/w), an amount of suspending agent between 5.0%> (w/w) and 35.0%> (w/w) and an amount of nutritional agent comprised between 0.5%> (w/w) and 40.0%> (w/w). Optionally, buffering, flavoring, edulcorating/sweetening and antioxidant agents can be comprised in the compositions in bags.

The edulcorating/sweetening agents for the preparation of the granulate composition for oral suspensions are chosen in the group comprising potassium acesulfame, sorbitol, mannitol, isomalt, maltitol, lactitol, xylitol, aspartame, cyclamic acid, cyclamate salts, lactose, sucralose, saccharine and saccharine salts or their mixtures.

According to a preferred embodiment of the present invention, the sweetening agent is chosen in the group consisting of sucralose, mannitol, sorbitol. More preferably, the sweetening agent is sucralose.

The compositions in the form of granulate in bags for oral suspensions comprise controlled release OKG granules in a percentage between 30% and 85% by weight, corresponding to an OKG amount of about 0.9 g and 3.0 g, together with excipients in the respective percentage amounts reported in Table 3.

Table 3

Component Percentage (%) (w/w)

Controlled release OKG granules 30.0 - 85.0

Lactoferrin 0.5 - 5.0

Mixture of xanthan gum, guar gum 5.0 - 25.0

and carob seed flour (Faramix^®)

Dehydrated orange juice 8.7 - 25.0

Orange flavor 0.5 - 5.0

Sucralose 0.0 - 2.5

Beta carotene 0.3 - 2.5 The compositions can be suspended in any chosen volume of aqueous solution, preferably comprised between 50 ml and 200 ml and, according to the chosen volume, the final compositions have different viscosity. When the described compositions are added to volumes smaller than 50 ml, the viscosity of the solution allows the composition administration by means of a spoon, and the composition is therefore useful for being administered to patients with swallowing problems.

Controlled release OKG granules can also be used for the preparation of solid forms, such as, for instance, tablets, lozenges, capsules.

The tablets are prepared by mixing controlled release OKG granules in an amount between 30% (w/w) and 50% (w/w), together with one or more excipients chosen among diluents, binders, disintegrants and lubricants.

The preferred tablet composition comprising controlled release OKG granules is reported in Table 4.

Table 4

Component Percentage (%) (w/w)

Controlled release OKG granules 30.0 - 48.9

Diluent(s) 35.0 - 50.0

Binder(s) 15.0 - 30.0

Disintergrant(s) 1.0 - 6.0

Lubricant(s) 0.1 - 1.2 The tablets can optionally comprise flavoring agents, coloring agents, antioxidant agents, buffering and antiseptic agents and can be coated by a filmogen coating.

The solid forms in tablets, lozenges or capsules can also comprise one or more nutritional substances having the same meaning reported for the bag compositions, in an amount between 0.1% (w/w) and 4.0% (w/w) by reducing the diluent amount.

Binders and diluents comprised in the OKG tablets have the same meaning as stated before.

The disgregant is chosen in the group consisting of sodium carboxymethylcellulose, also called carmelose sodium, cross-linked sodium carboxymethylcellulose, also called croscarmelose sodium, cross-linked polyvinylpyrrolidone, also called crospovidone, starch, pregelatmized starch, sodium starch glycolate or their mixtures.

The lubricant is chosen in the group consisting of silica, magnesium or calcium stearate, sodium stearyl fumarate, hydrogenated vegetable oils, mineral oils, polyethylene glycols, sodium lauryl sulphate, glycerides, glyceryl dibehenate, glycerol stearate or their mixtures.

According to a preferred embodiment of the present invention, the disgregant is chosen in the group consisting of sodium starch glycolate and croscarmelose sodium; the binder is microcrystalline cellulose; the diluent is chosen in the group consisting of mannitol and pregelatmized starch and their mixtures. Mixtures of mannitol and microcrystalline cellulose having diluting and binding properties are commercially available with the trademark Avicel^® HFE-12. The lubricant is chosen in the group consisting of glyceryl dibehenate and glycerol stearate. The present invention also describes the process for the preparation of controlled release OKG granules and the preparation of the compositions containing them.

The processes for the preparation of controlled release granules comprises the step of OKG granules preparation and the following step of coating step with polymers useful for obtaining controlled release of OKG. The obtainment of granules characterized for having homogeneous size is important for their complete and reproducible coating.

Wet granulation processes, dry granulation processes and fluid bed granulation processes for obtaining controlled release OKG granules are here described and compared in this invention.

Wet granulation processes are carried out in high-shear mixers wherein OKG powder in an amount between 30.0% (w/w) and 80.0%> (w/w) is mixed with binders in an amount between 5.0%> (w/w) and 15.0% (w/w) and diluents an amount between 4.5%) (w/w) and 20.0%> (w/w), and a wetting phase, generally water, is added to the solid mixture for obtaining a homogeneous wet mass. The obtained OKG granules are calibrated in a granulator equipped with net size between 1.14 and 2.5 mm, for a period of time comprised between 5 and 20 minutes. The granules are then dried, preferably in a fluid bed at a temperature comprised between 40°C and 80°C, in a range of time comprised between 30 minutes and 2 hours, with air flows comprised between 50 and 200 m³/h. The granules obtained by means of the aforesaid wet granulation processes have a size smaller than 1 mm in a percentage higher than 95% and a percentage of granules higher than 50%> has a size comprised between 250 μηι and 800 μηι. OKG granules are then coated for obtaining controlled release OKG. Comparative processes for the preparation of OKG granules are reported described in the present invention also with the use of the fluid bed apparatus.

An aqueous suspension, comprising binder in an amount between 20% (w/w) and 70% (w/w), is sprayed on the OKG powder in a weight ratio comprised between 30%) (w/w) and 80%> (w/w), with air flows comprised between 70 and 90 m³/h at a temperature comprised between 40°C and 80°C. In the fluid bed apparatus OKG adheres to the walls of the apparatus and even if the use of fluid bed has the advantage of using only one instrument for granule preparation and coating it is not utilizable and the wet granulation is preferable.

The second step of the process for obtaining controlled release OKG granules involves coating of the obtained granules by means of suitable polymers. OKG granules are coated in a fluid bed by spraying a solution comprising a binder in an amount between 5% (w/w) and 15% (w/w), a plasticizer in an amount between 0.5%> (w/w) and 5.0%> (w/w) and one or more polymers suitable for controlled release in an amount between 10%> (w/w) and 30%> (w/w).

Preferably, the polymers suitable for controlled release are natural polymers. The obtained suspension is kept under stirring for a time range comprised between 10 minutes and 1 hour. The suspension spraying speed on the granules is comprised between 10 g/min and 50 g/min, preferably between 10 g/min and 30 g/min, the air temperature in entrance is comprised between 40°C and 80°C, preferably between 55°C and 70°C, and the air flow in entrance is comprised between 60 m³/h and 120 m³/h.

The OKG granules, obtained by wet granulation process have homogeneous size, no agglomerate are formed and high yield, higher than 85% are showed. Moreover the operating condition of the process for obtaining the OKG granules avoid degradation of OKG and allow to obtain OKG granules having homogeneous size, important to obtain solid forms for medicinal preparation without limitations. The OKG granule are characterized to release quantitatively OKG at pH higher than 6.0.

The process for obtaining controlled release granules wherein the amount of OKG is in an amount from 30% to 80% in respect to the total granule weight, presents yields higher than 80%>, and it is able to overcome the technical problems of the state of the art. This invention overcome the technical problem of the prior art and provide the obtainment of the OKG controlled release comprising ah high quantity of active ingredient with high process yield.

Controlled release OKG granules are useful can be used for the preparation of pharmaceutical, nutritional or alimentary compositions without limitations.

The preparation of compositions for oral use, such as for instance granulates for oral suspensions in bags and tablets, is also described.

The process leading to the obtainment of granules in thermo-welded bags involves the mixing of the controlled release granules in Turbula mixers or V-blenders with the suitable excipients and, optionally, other active principles and/or nutritional substances.

Composition in form of tablets are obtained by means of direct compression of

OKG granules with the excipients and the instruments known in the art.

The compositions comprising controlled release OKG granules according to the present invention are useful in the treatment of patients suffering from malnutrition and/or malabsorption, since OKG improves appetite, body weight and, generally, the state of the health. In particular, OKG compositions are useful for particular catabolic states, like major burn cases or in surgically treated patients and in children suffering from growth delay.

The described compositions can also be taken by athletes or persons suffering from physical stress.

Patients and healthy subjects can assume the compositions comprising controlled release OKG granules in aqueous suspensions, tablets, capsules or simply dispersed in food, without limitations.

The compositions comprising controlled release OKG granules can be administered to patients and healthy subjects in a single or multiple dose, with a daily dosage comprised between 2 g and 20 g without any side effects.

The compositions comprising controlled release OKG granules, wherein a single dose comprises an OKG amount of 2 g, can be administered up to 10 times a day, or 3 single doses can be administered thrice a day, or 5 single doses can be administered twice a day, for a total daily dosage of 20 g OKG, without side effects and well tolerated by patients.

The described compositions in controlled release OKG granules can be prepared in bags, in glass or plastic bottles which are ready to use after having added aqueous solutions.

The compositions comprising controlled release OKG granules can be comprised in alimentary preparations also suitable for enteral use, and they can be administered by means of a nasogastric tube, a pharyngostomic tube, a gastrostomy tube or a digiunostomy tube.

The compositions comprising controlled release OKG granules can be part of a kit, comprising separate OGK composition ready for use. Examples 1-6 describe processes comparing the process for obtaining granule such as wet granulation and dry granulation fluid bed granulation, for the production of OKG granules.

Examples 2-6 are comparative examples useful to demonstrate that the process for the obtainment of granules results from optimal combination of chosen process parameters, excipients and amount thereof.

Example 1 describes the preparation of OKG granules by means of a wet granulation process wherein an OKG is mixed with microcrystalline cellulose and g hydroxy propyl cellulose in a high-shear. The obtained granulate has a water weight content of 1.03 % (w/w), the particle size analysis of obtained granules are comprised between 100 μηι and 1 mm, and in particular 66 % of them has a size comprised between 250 μηι and 800 μιτι, and 19% of them has a size comprised between 250 μηι and 63 μηι. The yield of the granulation process is 88.6 %.

Example 2 describes the preparation of granules by means of a wet granulation process in a high-shear granulator, with the same excipients of Example 1, but with a different amount. The obtained granulate is then dried in a fluid bed at 55 °C for 35 minutes, and the resulting granule has a water weight content of 1.01%>.The particle size analysis shows that the granules have a size comprised between 100 μηι and 1 mm, wherein 52% of them is comprised between 250 μηι and 800 μιτι; 42% of them is represented by granules having a size comprised between 250 μηι and 63 μιτι, and 11 ) of them has a size larger than 800 μηι.

The granulate obtained in Example 2 has a high percentage of particles having a size smaller than 250 μηι, thus resulting dusty and scarcely suitable for coating processes. Example 3 describes the process for the preparation of controlled release granules by means of wet granulation in a high-shear granulator, wherein the amount of excipients and granulation parameters are different in respect to Example 1.

The obtained granules after drying have water content of 1.03% and they have a size comprised between 100 μηι and 1 mm, wherein a percentage higher than 50%> is comprised between 250 μηι and 800 μηι and the yield is higher than 85%.

The granules obtained in Example 3 turns out to be more homogeneous if compared to the one obtained in Example 2, but the percentage of the granules having a size comprised between 250 μηι and 800 μιτι, useful for a good coating, is smaller than the ones of Example 1.

Example 4 describes the process for the preparation of OKG granules using a fluid bed apparatus equipped with top spray system. This process involves the preparation of an aqueous solution of pre-gelatinized starch which is sprayed on the OKG powder in fluid bed. OKG tends to adhere to the walls of the instrument even with strong incoming air flows, thus showing that OKG cannot be directly granulated in fluid bed.

Example 5 describes the preparation of OKG granules by means of a dry granulation process on nets having meshes sized 3.5 mm and 1 mm. The process involves the mixing OKG in an amount of 90%> (w/w) and pregelatinized starch in an amount of 10%> (w/w), which are directly loaded in the compactor feeder thus exerting increasing pressures.

The obtained granules has a size comprised between 100 μηι and 1 mm, wherein a percentage of 61 > has a size comprised between 250 μηι and 800 μηι and a percentage higher than 25% has a size comprised between 250 μηι and 63 μηι and they are not suitable for being coated. Example 7 describes the coating process of OKG granules, wherein an obtained aqueous solution of hydroxypropyl metal cellulose (HPMC), glycerol and natural polymer is sprayed in a fluid bed apparatus on OKG granules prepared according to Example 1 and the coated granules have homogeneous size comprised between 100 μηι and 1 mm, and in particular 88% of them has a size comprised between 250 μηι and 1 mm.

The final yield of the process for the preparation of the controlled release granules, comprising the step of preparation of OKG granules and the step of granules coating, is higher than 80%, and in particular is 81.5%.

In particular, the granules prepared according to Example 7 are suitable for being used for the preparation of pharmaceutical, nutritional, alimentary compositions in bags, in tablets or in capsules without limitations.

The granules can be used as they are or added to excipients without any limitation in order to prepare solid or liquid compositions.

For solid composition it is intent preparation in bags ready to be in aqueous solution, tablet or capsules.

OKG granules can be suspended in aqueous solution in bags ready for use. The Examples 8 to 12 describes the compositions comprising the OKG granules for different route of administrations.

Example 13 describes OKG release from the granules, prepared according to Example 7, in acid pH and neutral pH solutions, and the OKG amounts released in time are determined by means of HPLC chromatography analysis with spectropho to metric detection at 215 nm.

Example 14 and Example 15 describe a single centre, open trial on 42 elderly subjects to assess the acceptability and tolerability and safety of oral sachets comprising OKG controlled release granules in association with lactoferrin administered as 3 different mixtures of sachets in 50 ml of water or orange juice or in a yogurt pot, each one twice daily for 2 days.

No one of the forty-two elderly subjects receiving the oral composition comprising OKG controlled release granules in association with lactoferrin described in Example 9 complained about the taste or refused to receive the successive administration for all mixture in water, orange juice or in a yogurt pot, Furthermore the forty-two subjects showed an improvement of the body weight and Body Mass Index (BMI), and improvement of appetite.

The Example 15 describes the determination of the safety of the composition comprising OKG controlled release granules in association with lactoferrin wherein the evaluation was based on physical examination findings, vital signs measurements, recorded adverse events.

No alteration was shown with regard to vital parameters such as blood pressure, pulse body temperature and weight.

Some examples of the present invention are described hereinafter for exemplificative and not limitative purposes,

Example 1

Preparation of OKG granules by means of a wet granulation process in a high-shear granulator

The process described in this example for the preparation of OKG granules comprises the wet granulation of OKG in a high-shear granulator (Hi-Shear Mixer) and the drying of granules in fluid bed.

OKG in an amount corresponding to 2695 g was mixed in a high-shear granulator, like a Colette Gral 25 granulator, together with 560 g microcrystalline cellulose and 245 g hydroxypropyl methylcellulose, in the percentages as reported in Table 5.

Table 5

The solid components were mixed for about 30 seconds and then 1120 ml water were added, under stirring, and the granulation was maintained for further 4 minutes. The granules were calibrated by means of an oscillating granulator, like an Erweka AR 400, on a net whose meshes have a size of 1.14 mm, and then they were dried in fluid bed according to the parameters reported in Table 6. Table 6

The dried granules have a water content of 1.03% demined by Karl Fisher.

The particle size analysis of the obtained OKG granules was carried out by means of a Retsch vibrosieve by using sieves whose meshes have a size comprised between 63 μηι and 1 mm, with a stirring time of 10 minutes.

Table 7 reports the granules size and the granules relative percentages. Table 7

The yield of the described process corresponds to 88.6%.

The obtained granules may be then coated with controlled release polymers.

Example 2

Preparation of OKG granules by means of a wet granulation process in a high-shear granulator (Comparative example)

The preparation process of OKG granules comprises the wet granulation of OKG in a high-shear granulator (Hi-Shear Mixer) and the drying of the granules in fluid bed.

An OKG amount corresponding to 2695 g was mixed in a Colette Gral 25 granulator for about 60 seconds together with 665 g microcrystalline cellulose and 140 g HPMC. A volume of 1050 ml water was added to homogenize the mixture and the resulting mass is granulated for 3 minutes. The granules were calibrated by means of an oscillating granulator on a net meshes with size of 2.5 mm.

Table 8 reports the percentages of the components of the OKG granule.

Table 8

The wet granulate was dried in fluid bed with incoming air at 100 m³/h at 55 °C for 35 minutes.

The particle size analysis of the obtained granules comprising OKG was carried out by means of a Retsch vibrosieve by using sieves whose meshes size comprised between 63 μηι and 1 mm, with a stirring time of 10 minutes.

Table 9 reports the results of the particle size analysis.

Table 9

./. follows Dried granules size Granules Percentage (%)

Between 500 μηι and 250 μηι 27.44

Between 250 μηι and 125 μηι 21.89

Between 125 μηι and 63 μηι 11.81

<63 μηι 2.35

Total 100

The obtained granules have a water weight content corresponding to 1.01%, determined by Karl Fisher.

The granules were not coated because a portion of granules larger than 45% has a size smaller than 250 μηι and is too dusty.

Example 3

This example describes process for the preparation of granules comprising an OKG comprising the step of wet granulation in a high-shear granulator (Hi-Shear Mixer) and the drying of the granules in fluid bed.

An OKG amount corresponding to 2695 g was mixed with 665 g microcrystalline cellulose and 140 g HPMC in a Colette Gral 25 granulator and the mixture was mixed for 30 seconds. 1050 ml water have been added under stirring, and the wet mass is granulated for about 7 minutes. The granules were calibrated by means of an oscillating granulator on a net meshes with size corresponding to 2.5 mm and then to 1.14 mm.

The granule composition is reported in Table 10.

Table 10

The wet granulate was dried in fluid bed with air flow of 100 m³/h at 55 °C for 35 minutes.

The particle size analysis of the obtained granules comprising OKG was carried out by means of a Retsch vibrosieve by using sieves whose meshes have a size comprised between 63 μηι and 1 mm, with a stirring time of 10 minutes.

Table 11 reports the results of the particle size analysis.

Table 11

./. follows Between 800 μηι and 500 μηι 32.28

Between 500 μηι and 250 μηι 27.25

Between 250 μηι and 125 μηι 11.5

Between 125 μηι and 63 μηι 8.98

<63 μηι 3.92

Total 100

A percentage of granules higher than 55% has a size comprised between 250 μηι and 800 μηι. The granules have then been coated with controlled release polymer as described in Example 7.

The percentage of the granules having a size suitable for a good coating, is smaller than the ones of Example 1.

The yield of the coating process corresponds to 87%, with a final water content of 1.08%), determined by Karl Fisher.

Example 4

Preparation of OKG granules by means of a fluid bed granulation (Comparative example)

An amount of pregelatinized starch (PGS) corresponding to 180 g was dissolved in 1620 ml water and the obtained solution is sprayed on 1500 g OKG in a fluid bed apparatus provided with a Top Spray system with an incoming air flow of 80 m³/h at 60°C. OKG adhered to the apparatus walls and the granulation process was suspended. Example 5

Preparation of OKG granules by means of a dry granulation process (Comparative example)

An OKG amount corresponding to 1500 g was mixed with 166.7 g PGS in a V- blender for 15 minutes at 16 rpm. The mixture was directly loaded into the feeder of the Alexanderverk WP50/N75 compactor, provided with nets whose meshes have a size comprised between 3.15 mm and 1 mm. The mixture was processed in three steps, wherein in the first step the components were mixed exerting no pressure on the rolls, then the pressure was increased to 25 bar pressure, and at the end the pressure was increased to 100 bar.

Table 12 reports the granules composition.

Table 12

The particle size analysis of the obtained granules comprising OKG was carried out by means of a Retsch vibrosieve by using sieves meshes with size comprised between 63 μηι and 1 mm, with a stirring time of 10 minutes.

The results of the granulometric analysis of the granules obtained by dry compaction are reported in Table 13. Table 13

The OKG granule obtained are not suitable for being coated.

Example 6

The example describes process for the preparation of granules comprising the step of wet granulation in a high-shear granulator (Hi-Shear Mixer) and the drying of the granules in fluid bed. An OKG amount corresponding to 1500 g and 150 g powdered PGS were mixed and 420 ml water are added to the resulting mixture. The granulation was continued for about 20 minutes. At the end of the granulation, the obtained product is sieved on a net whose meshes have a size corresponding to 4 mm in order to get rid of the large agglomerates. The granules were calibrated by means of an oscillating granulator on a 1.14 mm net.

Table 14 reports the granules composition.

Table 14

The granules were dried in fluid bed with air flow of 120 m³/h at 55°C for 22 minutes. The final water content corresponds to 0.61%, determined by Karl Fisher.

The particle size of the obtained granules comprising OKG was carried out by means of a Retsch vibrosieve by using sieves whose meshes have a size comprised between 63 μηι and 1 mm, with a stirring time of 10 minutes.

The results of the particle size analysis are reported in Table 15, which shows that a granule percentage higher than 55% has a size comprised between 250 μηι and 800 μπι. Table 15

This process leads to the formation of large-sized agglomerates and particles, and their removal influences the final yield of the granulation process which is then lower than 80%.

Example 7

Preparation of controlled release OKG granules

The solution for coating the granules was prepared by adding 40 g hydroxypropyl methylcellulose (HPMC) to 752 ml water, previously heated at 60°C and kept under stirring until reaching complete HPMC dissolution. 2240 ml water, 1376 g of 25% solution of shellac natural polymer (Aquagold ) and 16 g glycerol.

The solution was kept under stirring for 20 minutes before starting the coating step.

The solution was sprayed on OKG granules obtained from Example 1 in a fluid bed apparatus according to the parameters reported in Table 16.

Table 16

Table 17 reports the components comprised in the controlled release OKG granules and their relative percentages.

Table 17

Component Amount Percentage (%)

(g) (w/w)

OKG granules 800 66.67

Natural polymer 1376 28.67

(Aquagold^®, 25% solution)

HPMC 40 3.33

Glycerol 16 1.33 The coated granules were dried in fluid bed apparatus with 35°C for 15 minutes and 1100 g controlled release OKG granules are obtained.

The yield of the coating process was of 91.4% and the yield of the total process for obtaining controlled release granules was of 81.5%.

The water weight content of controlled release OKG granules corresponds to

1.07%.

The results of the particle size analysis are reported in Table 18.

Table 18

Dried coated granules size Granules percentage

(%)

> 2 mm 0.00

Between 2 mm and 1 mm 2.9

Between 1 mm and 800 μηι 11.84

Between 800 μηι and 500 μηι 30.03

Between 500 μηι and 250 μηι 43.11

Between 250 μηι and 125 μηι 8.50

Between 125 μηι and 63 μηι 3.40

< 63 μηι 0.20 Example 8

Preparation of a composition comprising controlled release OKG granules in bags

An amount corresponding to 3895 g controlled release OKG granules prepared according to Example 7 was mixed in a Turbula mixer for 10 minutes with 50 g lactoferrin and 900 g suspending agent, Faramix^®. The mixture is then aliquoted in bags, wherein each bag contains 2 g OKG.

Table 19 reports the unitary composition of each bag.

Table 19

Example 9

Preparation of compositions comprising OKG granules for aqueous suspensions

An amount corresponding to 3985 g controlled release OKG granules prepared according to Example 7, was mixed in a Turbula mixer for 10 minutes together with 50 g lactoferrin and excipients, orange juice, orange flavor, sucralose and beta carotene.

Table 20 reports the unitary amounts of the composition. Table 20

The mixture was then aliquoted in thermo-welded bags to be suspended in aqueous solutions.

Bags comprising an OKG amount corresponding to 2 g can be poured in a volume of aqueous solution comprised between 50 ml and 100 ml by mixing for about 30 seconds and settling for about 2 minutes, until the mixture acquires a gelatinous consistency.

The composition can be dispersed in a volume corresponding to 100 ml of aqueous solution, or 200 ml for obtaining a less viscous consistency.

Example 10

Preparation of a semisolid food comprising controlled release OKG granules

An amount of controlled release OKG granules prepared as described in Example 7, corresponding to 19 g and comprising an OKG amount corresponding to 10 g, was dispersed in semisolid food, by mixing with the help of a spoon until reaching a complete dispersion of the granules. Food can be administered by means of a spoon or kept at a temperature of 4°C. Example 11

Preparation of tablets comprising controlled release OKG granules

The controlled release OKG granules prepared as described in Example 7 weres mixed with mannitol, pregelatinized starch, microcrystalline cellulose, sodium starch glycolate, previously sieved on a net whose meshes have a size of 0.8 mm. Glyceryl dibehenate was added to the homogeneous mixture.

The composition is reported in Table 21.

Table 21

The homogeneous mixture was loaded in a Kilian rotary compressor provided with a round concave punch having a 12 mm diameter. The mixture was compressed by applying a 10-20 KN compression force. Example 12

Preparation of tablets comprising controlled release OKG granules

The controlled release OKG granules prepared as described in Example 7 were mixed with a mixture of mannitol and microcrystalline cellulose (Avicel^®), glycol glycerol stearate (Precirol^®) and croscarmelose, previously sieved on a net whose meshes have a size of 0.8 mm. The homogeneous mixture was loaded in a Kilian rotary compressor provided with a round concave punch having a 12 mm diameter. The mixture is compressed by applying a 10-20 KN compression force.

The unitary composition of the tablets is reported in Table 22.

Table 22

Example 13

Determination of the dissolution of OKG controlled release granules.

The dissolution of the OKG granules, as prepared in Example 7, was determined in solutions at pH 1 and pH 6.8, and the amount of released OKG was determined by means of chromatographic analysis, HPLC.

An amount corresponding to 3.9 g of granules prepared according to Example 7, comprising 2 g OKG, was placed in 750 ml of a HC1 solution 0.1 M. The same amount of granules was then placed in 1000 ml aqueous solution brought to pH 6.8 by adding NaOH 1 M. The OKG amount released by the two solutions was determined by means of chromatographic analysis HPLC.

The reverse-phase HPCL chromatographic analysis involves the use of a C8 column sized 250 x 4.6 mm, with a particle size of 5 μηι and a spectrophotometric revelation at 215 nm.

The OKG elution occurs in linear gradient in 15 minutes from a H₃PO₄ solution 0.02 M to a methanol solution, with a 0.6 ml/min flow. L-ornithine was eluted at a retention time (RT) of 3.8 minutes and a-ketoglutaric acid was eluted at a RT of 10.5 minutes.

The OKG amount released in time in the solution at acid pH and neutral pH is reported in Table 23.

The OKG amount at acid pH at 15 minutes, 30 minutes and 45 minutes is under the limit of instrumental determination.

Table 23

The OKG released from granules in acid environment is lower than 15% after about two hours, whereas at pH 6.8 the release is higher than 40% after 15 minutes and becomes higher than 95% after 45 minutes. Example 14

Determination of the acceptability and tolerability after the administration of the composition of Example 9 containing OKG-LF

The composition as described in Example 9 in sachets was administered in a single centre, open study to assess the acceptability and tolerability of oral OKG-LF sachets administered as 3 different mixtures of OKG-LF sachets in 50 ml of water or orange juice or in a yogurt pot, each one twice daily for 2 days in elderly subjects with anorexia and protein energy malnutrition.

The primary endpoint was the acceptability of 3 different mixtures of OKG-LF sachets in 50 ml of water or orange juice or in a yogurt pot.

The secondary endpoints were the evaluation of the rate of the mixture preparation taken; the relative improvement of anthropometric indexes such as body weight and Body Mass Index (BMI), and the relative improvement of asthenia and appetite according to 10-point Visual Analogue Scales (VAS).

In the clinical study 42 elderly subjects, 19 male and 23 female with average age of 68 years, were enrolled; 15 subjects were affected by anorexia defined as a declined food intake declined over the past 3 months due to loss of appetite, and 27 subjects had protein-energy malnutrition, defined as Mini Nutritional Assessment score < 23,5. All subjects had not taken new treatment nor food supplement containing anabolic amino acids and/or lactoferrin in the last 2 weeks.

A total of 42 elderly subjects were administered to OKG-LF sachets for 6 days by oral route, as 3 different mixtures of OKG LF sachets in 50 ml of water or orange juice or in a yogurt pot, each one twice daily for 2 days; subjects were randomized 1:1:1 :1:1 :1 according to one of the following six schemes of fluid administration: Scheme Days 0-1 Days 2-3 Days 4-5

A Water Orange juice Yogurt

B Orange juice Water Yogurt

C Yogurt Orange juice Water

D Water Yogurt Orange juice

E Orange juice Yogurt Water

F Yogurt Water Orange juice

At Visit 1 - Screening Visit (Day 0) the subjects were weighed, height was measured, malnutrition was assessed according to the Mini Nutritional Assessment^®, asthenia and appetite according to 10 point Visual Analogue Scales. Drawing of blood samples for laboratory test were performed Eligible subjects received the composition of Example 9 in a kit box for 6 days. Acceptability concerning the first mixture was evaluated according to Taste Panel Evaluation within 5 minutes after each administration. Subjects were provided also with a diary.

At Visit 2 (Day 2) the acceptability of the second mixture was evaluated according to Taste Panel Evaluation within 5 minutes after each administration.

At Visit 3 (Day 4) the acceptability of the third mixture was evaluated according to Taste Panel Evaluation within 5 minutes after each administration.

The Final Visit 4 at the End of Treatment (Day 6) occurred the day after the last taking of the composition. The composition of Example 9 accountability was checked and subject's diary was reviewed. During the visit the subjects were weighed, height was measured, malnutrition was assessed according to the Mini Nutritional Assessment^®, asthenia and appetite according to 10 point Visual Analogue Scales. Drawing of blood samples for laboratory test was performed. The acceptability of the 3 mixtures was assessed by Taste Panel Evaluation Form, which calls for the subject to rate the three samples for appearance, colour, odour, flavour, texture, and overall liking, using five point hedonic scales (from worst to best). The subjects indicated also whether the sample is acceptable (yes, no).

None of the forty- two elderly subjects receiving the OKG-LF composition described in Example 9 complained about the taste or refused to receive the successive administration for all mixture in water, orange juice or in a yogurt pot. The taste panel evaluation comprised: appearance colour, odour, flavor, texture.

The secondary Efficacy Parameters, such as the rate of the mixture preparation taken and the relative improvement of anthropometric indexes, weight, BMI and improvements of asthenia and appetite according to 10 point Visual Analogue Scales were calculated between baseline (VI) and the end of treatment phase (V4).

The forty- two subjects showed an improvement of the body weight and Body Mass Index (BMI), and improvement of appetite of 2-3 average point Visual Analogue Scales in 100% of subjects.

Example 15

Determination of the safety of the OKG-LT composition of Example 9 in sachets.

42 Eligible subjects received OKG-LF sachets for 6 days by oral route, as 3 different mixtures of OKG LF sachets in 50 ml of water or orange juice or in a yogurt pot, each one twice daily for 2 days. The Safety evaluation was based on physical examination findings, vital signs measurements, recorded adverse events.

No alteration was shown with regard to vital parameters such as blood pressure, pulse and body temperature.

Claims

1. A composition comprising ornithine alpha-ketoglutarate granules, wherein the granules comprise an amount of ornithine alpha-ketoglutarate between 30% and 80%> (w/w) with respect to the final weight of granules together with acceptable excipients.

2. The composition according to claim 1, wherein ornithine alpha- ketoglutarate granules comprise:

- Ornithine alpha-ketoglutarate: 30.0 - 80.0%> (w/w);

- Diluent: 4.5 - 20.0% (w/w);

- Binder: 5.0 - 15.0% (w/w);

- Plasticizer: 0.5 - 5.0% (w/w);

- Controlled release polymer: 10.0 - 30.0%> (w/w), with respect to the final weight of granules.

3. The composition according to claim 2, wherein ornithine alpha- ketoglutarate granules comprise:

- Ornithine alpha-ketoglutarate: 45.0 - 80.0%> (w/w);

- Microcrystalline cellulose: 4.5 - 20.0%> (w/w);

- Hydroxypropyl methylcellulose: 5.0 - 15.0% (w/w);

- Glycerol: 0.5 - 2.5% (w/w);

- Shellac: 10.0 - 30.0%> (w/w), with respect to the final weight of granules.

4. The composition comprising ornithine alpha-ketoglutarate granules according to claim 1, comprising one or more nutritional ingredients and/or one or more active ingredients or alimentary and pharmaceutical acceptable excipients.

5. The composition according to claim 4, wherein the nutritional agent is chosen in the group consisting of milk proteins, vitamins, amino acids or their mixtures.

6. The composition according to claim 5, wherein the nutritional agents are milk proteins.

7. The composition according to claim 6, wherein the nutritional agent is lactoferrin in an amount comprised between 10% (w/w) and 70% (w/w) with respect to the total weight of the composition.

8. The composition comprising ornithine alpha-ketoglutarate granules according to claim 1 in form of tablets, capsules, lozenges or granulate in thermo- welded bags.

9. The composition according to claim 8 in thermo-welded bags, comprising an amount of ornithine alpha-ketoglutarate granules comprised between 30%) (w/w) and 95% (w/w), nutritional agents comprised between 0.5%> (w/w) and 40%) (w/w), suspending agents comprised between 5% (w/w) and 35% (w/w) with respect to the total weight of the composition and, optionally, flavoring, sweetening, buffering, antioxidant and preservative agents.

10. The composition in thermo-welded bags according to claim 9 comprising:

- Ornithine alpha-ketoglutarate granules: 30.0 - 80.0% (w/w);

- Lactoferrin: 0.5 - 5.0% (w/w);

Suspending agent (xanthan gum, guar gum, carob seed flour): 5.0

(w/w);

- Dehydrated orange juice: 8.7 - 25.0% (w/w);

- Orange flavor: 0.5 - 5.0% (w/w);

- Sucralose: 0.0 - 2.5% (w/w); - Beta carotene: 0.3 - 2.5% (w/w), with respect to the total weight of the composition.

11. The composition according to claim 8 in the form of tablets, wherein the amount of ornithine alpha-ketoglutarate granules, according to claim 1, is comprised between 30%> (w/w) and 50% (w/w) with respect to the weight of tablet together with alimentary or pharmaceutically acceptable excipients.

12. A process for the preparation of ornithine alpha-ketoglutarate controlled release granules comprising an amount of ornithine alpha-ketoglutarate between 30%> (w/w) and 80%> (w/w) with respect to the final weight of granules comprising the step of:

- mixing ornithine alpha-ketoglutarate in an amount comprised between 30%> (w/w) and 80%> (w/w) with a binder in an amount comprised between 5% (w/w) and 15%) (w/w) and a diluent in an amount comprised between 4.5% (w/w) and 20.0% (w/w) with respect to the total weight of the granules;

- adding a volume of aqueous solution in an amount comprised between 1000 ml and 1500 ml;

- granulating the mixture in a high-shear mixer;

- drying the granules at temperature comprised between 40°C and 80°C in a time range higher than 30 minutes; and

- coating the dried granules with aqueous suspension comprising a polymer in a fluid bed apparatus, wherein the an air flow in entrance is between 60 and 120m³/h, spray rate between 10 and 50 g/min and air temperature comprised between 40°C and 80°C.

13. A process for the preparation of thermo-welded bags comprising ornithine alpha-ketoglutarate controlled release granules, according to claim 1 , wherein the granules are mixed in a V-blender with pharmaceutical acceptable excipients, one or more nutritional agents and/or active principles and then administered.

14. A process for the preparation of tablets comprising ornithine alpha- ketoglutarate granules, according to claim 1, comprising an amount of controlled release granules between 30% (w/w) and 50% (w/w) with respect to the tablet weight, together with pharmaceutical acceptable excipients and the resulted mixture is compressed in a tableting machine.

15. A compositions comprising controlled release ornithine alpha- ketoglutarate granules, wherein said granules comprise an amount of ornithine alpha- ketoglutarate between 30%> and 80%> (w/w) with respect to the final weight of granules, useful in the treatment and/or the prevention of malnutrition and/or malabsorption state.

16. A composition comprising controlled release ornithine alpha- ketoglutarate granules, according to claim 1, for the treatment or prevention of pathologies related to states of malnutrition and/or malabsorption and/or physical stress.

17. Food supplement comprising controlled release ornithine alpha- ketoglutarate granules according to claim 1, wherein the total amount of ornithine alpha-ketoglutarate is comprised between 2 g and 20 g.