EA043776B1 - NUTRIENT COMPOSITION FOR BUILDING A DIET FOR PHENYLKETONURIA AND METHOD OF OBTAINING - Google Patents

Description

Field of technology to which the invention relates

The embodiments described herein relate to a nutritional composition for constructing a diet for phenylketonuria, that is, for the treatment of phenylketonuria, and a corresponding production method. The composition in accordance with the embodiments described herein can be used, for example, to prepare a powder product to be dissolved in a liquid food, drink or the like, such as water, before consumption, that is, a powder to be reconstituted in liquid. In addition, the composition in accordance with the embodiments described herein can also be used, for example, to prepare a liquid product that is ready to drink, that is, ready to drink. In addition, the composition in accordance with the embodiments described herein can, for example, be used as an ingredient for the manufacture of other food products, such as dairy products, in particular yogurt, or confectionery products, such as creams or puddings, baked goods , such as bread, crackers, breadsticks, snacks, cookies, cakes, sweet snacks, confectionery and pasta, etc., or other food beverages.

Prior Art

Phenylketonuria (PKU) is known to be a rare metabolic disease caused by impaired phenylalanine metabolism. Deficiency of the enzyme phenylalanine hydroxylase or its cofactor tetrahydrobiopterin causes abnormal accumulation of phenylalanine in body fluids and in the central nervous system, which appears to be the part most affected by this pathology. Depending on the residual ability to metabolize phenylalanine and therefore the concentration of this amino acid in the blood, the disease is classified as severe, moderate or mild. If phenylalanine concentrations in the blood are not controlled, patients experience cognitive deficits and irreversible mental retardation.

The average incidence of PKU among Europeans is 1:10,000/15,000, with a peak in Turkey (1:2600) and a very low incidence in Finland (1:200,000).

Once PKU is diagnosed, a more or less restrictive protein-free diet should be started depending on the patient's residual enzyme activity, and protein intake should be supplemented with specialty foods that do not contain phenylalanine. These foods are typically in powder form, reconstituted with water, or liquid foods, ready to eat, and consist of free amino acids, possibly carbohydrates and fats, mineral salts, trace elements and vitamins. The protein component typically consists of all amino acids except phenylalanine, and the amino acid profile is tailored according to the age group for which the particular product is intended. It is important that patients consume these foods because they represent the only safe source of protein, that is, completely free of phenylalanine and containing sufficient amounts of tyrosine and other essential amino acids.

Specialty products traditionally used to treat PKU have several negative aspects:

organoleptic aspect: amino acids give foods an unpleasant taste and odor, such as, for example, sour, bitter, sulfur or cabbage taste or odor. Although patients become accustomed to these unusual tastes from infancy, there is a need to improve the sensory properties of these products, as well as to diversify the choice of products;

absorption of amino acids at the gastrointestinal level: unlike the absorption of normal proteins, individual amino acids are absorbed much more quickly at the intestinal level, since the protein hydrolysis step is not necessary. This leads to differences in nitrogen retention at body level and a decrease in the feeling of fullness (in many patients this correlates with the risk of excess weight).

In addition, the amino acids used to make these products are obtained through hydrolysis of natural proteins or through chemical synthesis. Thus, formulations have a synthetic or artificial appearance in the eyes of consumers, and manufacturers try to limit this to the necessary minimum.

Since the 1990s, the possibility of using a specific peptide to treat PKU with food has been discovered. A peptide called glycomacropeptide (GMP) is produced by the breakdown of K-casein by chymosin during cheese production and can be easily isolated from whey. GMP is suitable for food-based treatment of PKU because it is naturally low in phenylalanine, so it can be included in specialty foods as a source of protein.

The main positive aspects of GMP for the treatment of PKU are the following:

GMP is a peptide very similar to natural proteins and has a neutral taste, without the unpleasant aftertaste associated with amino acids. This characteristic allows the development of food products that have a better sensory profile compared to traditionally used amino acid mixtures that do not use GMP. GMP-containing products help patients adhere to their diet better by improving taste and increasing the amount of food they eat.

- 1 043776 va products suitable for protein supplements;

improvement of certain aspects related to the absorption of the peptide: hydrolysis of the peptide ensures the absorption of amino acids, which occurs more slowly and physiologically, thereby increasing the feeling of satiety in the patient. In addition, protein retention is increased and fluctuations in phenylalanine levels are reduced. Thus, this last aspect improves phenylalanine control in the blood compared to a diet that provides only amino acids;

Significant levels of LNAA (Large Neutral Amino Acid) compounds are found in GMP, a family of amino acids that share the same transporter as phenylalanine at the level of the blood-brain barrier. This family includes leucine, valine, isoleucine, histidine, methionine, tryptophan, tyrosine and threonine. It is possible to use mixtures of these amino acids that will saturate these transporters at the brain level, limiting the flow of toxic phenylalanine into the brain. Therefore, the use of LNAA mixtures to treat patients with phenylketonuria may reduce the concentration of phenylalanine accumulated at the brain level, which is neurotoxic. Therefore, LNAAs present in GMP may play a protective role at the brain level;

a diet containing GMP foods stimulates the growth of patients with phenylketonuria, promoting an increase in muscle mass and a decrease in fat mass.

It is also known that GMP is not completely free of phenylalanine, and this limits the amount of peptide that can be introduced into products. Despite this limitation, the amount of peptide that can be used in various products is sufficient to significantly improve their nutritional and organoleptic aspects, factors that positively influence patient compliance with dietary therapy. Moreover, the current recommendation from nutritionists is that GMP can be used to replace about 50% of protein intake to enhance the nutritional value and palatability of the diet.

Glycomacropeptide (GMP) is also found at low levels in other amino acids such as arginine, cystine, histidine, tryptophan, tyrosine and leucine. Among these amino acids, histidine, tryptophan and leucine are essential, arginine, cystine and tyrosine are essential during the child's developmental phase. Tyrosine also becomes an essential amino acid throughout the lives of people with PKU because it cannot be produced from phenylalanine as it normally does. Therefore, non-GMP amino acids must be supplemented to meet the nutritional needs of patients with PKU.

WO-A-2010/144821 describes a GMP-based nutritional product for dietary therapy of PKU and other metabolic disorders, which, in addition to the GMP, includes additional amounts of arginine, leucine and optionally other amino acids such as tyrosine.

WO-A-2015/205489 describes a nutritional composition in powder form that can be combined with water to prepare a milk-like drink for PKU diet therapy. The powder formulation contains casein-GMP (cGMP) and complementary essential amino acids, which are a mixture of tyrosine, arginine, tryptophan, leucine, histidine, which in combination provide the desired amino acid profile.

US-B-9,414,619 describes a ready-to-use liquid nutritional formulation for the treatment of PKU, tyrosinemia, or other metabolic errors. The formulation contains cGMP as the primary protein source and complementary essential amino acids to complete the required protein profile. The drink also contains vitamins and minerals to meet daily requirements for these elements, and when used as a balanced diet, the composition also includes a source of carbohydrates and a source of fat.

WO-A-98/08402 describes a nutritional formula for patients suffering from PKU that contains a mixture of casein-GMP (cGMP) and complementary essential amino acids other than phenylalanine as a protein source.

US-B-6,355,612 describes a low digestion rate protein product for the manufacture of an enteral composition that modulates plasma postprandial amino acid levels.

Document FR-A-3.002.831 describes a ready-to-eat food product for patients with metabolic pathology of one or more amino acids.

Also known is the publication Position of the American Dietetic Association: Use of Nutritive and Nonnutritive Sweeteners, Journal of the American Dietetic Association, Vol. 104, no. 2, 1 February 2004, pages 255-275.

One disadvantage of the known nutritional formulations is that although they provide GMP and complementary essential amino acids for dietary therapy purposes in PKU, they nevertheless have inadequate sensory properties that make them sensory unpleasant to those taking the formulations described above.

Therefore, there is a need to improve the nutritional composition for constructing a diet for phenylketonuria and the corresponding production method that can overcome the

- 2 043776 at least one of the disadvantages of the prior art.

In particular, one of the objects of the present invention is to provide nutritional compositions that, in addition to providing GMP and complementary essential amino acids for dietary therapy purposes in PKU, have adequate sensory properties to be organoleptically pleasing to those taking the formulations described above.

Applicant has developed, tested and implemented the present invention to overcome the shortcomings of the state of the art and to achieve these and other objects and advantages.

The essence of the invention

The present invention is set forth and characterized by independent claims, while dependent claims describe other features of the invention or variations of the basic idea of the invention.

In accordance with the above objectives, embodiments relate to a nutritional composition for constructing a diet for phenylketonuria, in particular useful for the treatment of phenylketonuria. In one embodiment, the nutritional composition contains a glycomacropeptide (GMP) as a primary source of proteins and additional complementary essential amino acids to complete the desired protein profile, supplied in a mixture containing arginine, cystine, histidine, tryptophan, tyrosine and leucine. The nutritional composition contains arginine ranging from 33 to 50 milligrams arginine/gram equivalent protein and leucine ranging from 47 to 76 milligram leucine/gram equivalent protein.

In some embodiments, which can be combined with all of the embodiments described herein, the nutritional composition has a tyrosine content of from 112 to 188 milligrams of tyrosine/gram of equivalent protein.

In certain embodiments, which may be combined with all of the embodiments described herein, in a nutritional composition the total weight of the additional complementary essential amino acids is from 35% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids amino acids.

In some embodiments, which may be combined with all of the embodiments described herein, the nutritional composition has a tryptophan content of 12 to 22 milligrams tryptophan/gram equivalent protein.

In some embodiments, which can be combined with all embodiments described herein, the nutritional composition has a histidine content of 22 to 37 milligrams histidine/gram equivalent proteins.

In some embodiments, which can be combined with all of the embodiments described herein, the nutritional composition has a cystine content of 3.8 to 6.2 milligrams cystine/gram equivalent protein.

According to some embodiments, which can be combined with all of the embodiments described herein, arginine supplied as a complementary essential amino acid, in addition to GMP-derived arginine, is derived from arginine supplied in the form of an arginine salt, e.g. , arginine aspartate salts.

Additional embodiments described herein relate to a food or beverage product containing a composition as disclosed herein.

In accordance with some embodiments, which can be combined with all embodiments described herein, the food or beverage also contains carbohydrates and fats.

In some embodiments, which can be combined with all of the embodiments described herein, carbohydrates provide 20% to 50% of the energy of the food or beverage.

In some embodiments, which can be combined with all embodiments described herein, fats provide from 1% to 50% of the energy of the food or beverage.

In accordance with some embodiments, which can be combined with all embodiments described herein, the weight ratio between proteins and carbohydrates in the food or beverage that includes the nutritional composition is from 0.85 to 2.5.

In accordance with some embodiments, which can be combined with all embodiments described herein, the weight ratio between proteins and fats in the food or beverage that includes the nutritional composition is from 10 to 130.

In some embodiments, which can be combined with all embodiments described herein, the carbohydrates in the food or beverage contain trehalose, and the trehalose content is from 35% to 95% by weight, based on the total amount by weight of carbohydrates present.

Other embodiments described herein relate to a method for making

- 3 043776 nutritional composition for constructing a diet for phenylketonuria. In accordance with one embodiment, the method includes mixing a glycomacropeptide (GMP) as the main source of proteins and additional complementary essential amino acids to complete the desired protein profile, supplied in a mixture containing arginine, cystine, histidine, tryptophan, tyrosine and leucine. In one embodiment, arginine and leucine are provided in quantities such that the nutritional composition contains 33 to 50 milligrams of arginine/gram of equivalent protein and 47 to 76 milligram of leucine/gram of equivalent protein of leucine.

Other embodiments described herein relate to a method of making a food or beverage product for constructing a diet for phenylketonuria. In some embodiments, this method provides and mixes together at least: a nutritional composition as described herein, carbohydrates, and fats. In the case of producing a drink, the method involves mixing the nutritional composition, carbohydrates and fats with an edible liquid, in particular water.

These and other aspects, features and advantages of the present disclosure will be better understood by reference to the following description and the accompanying claims.

Various aspects and characteristics described in the present invention can be used individually, where possible. These individual aspects, such as aspects and characteristics in the specification or in the appended dependent claims, may be the subject of divisional applications.

It should be understood that if any aspect or characteristic that is discovered during the patenting process is already known, it should not be claimed and should be subject to a disclaimer.

Detailed Description of Some Embodiments

Various embodiments of the present invention will now be discussed in detail. Each example is presented to illustrate the invention and should not be construed as limiting it. For example, features shown or described to the extent that they are part of one embodiment may be adapted or related to other embodiments to create another embodiment. It is understood that the present invention includes all such modifications and variations.

Before describing these embodiments, it should also be made clear that the present invention is not limited in its application to the detailed description of the structure and arrangement of components as set forth in the following description. The present disclosure may provide other embodiments and may be obtained or performed in various other ways. It should also be made clear that the phraseology and terminology used herein are for descriptive purposes only and should not be construed as limiting.

Unless otherwise defined, all technical and scientific terms used herein and hereinafter have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention relates. Even though methods and materials similar or equivalent to those described herein can be used in the practice and testing of the present invention, such methods and materials are described hereinafter by way of example. In the event of a conflict, this application, including its definitions, will control. The materials, methods, and examples are for illustrative purposes only and should not be construed as limiting.

In general, all percentages and ratios reported refer to the weight of the total composition (w/w), unless otherwise indicated.

All percentage ranges shown herein are based on the assumption that the sum relative to the total composition is 100% unless otherwise stated.

It should be understood that all ranges stated herein include extreme values, including those that indicate a range between two values, unless otherwise noted.

The present description also includes ranges that are formed by overlapping or combining two or more of the described ranges, unless otherwise indicated.

This description also includes ranges that may be formed by combining two or more values taken at different points, unless otherwise noted.

The embodiments described herein relate to a phenylketonuria (PKU) diet nutritional composition having a nutritional profile of GMP protein and additional complementary amino acids to ensure balanced intake for the PKU diet.

In particular, the nutritional profile, that is, the composition of essential amino acids within a nutritional composition, may be expressed herein in milligrams per gram equivalent protein (g EP or gram equivalent protein).

Total equivalent proteins (EP) can be calculated herein as the sum

- 4 043776 GMP protein and the sum of additional complementary amino acids divided by 1.2, according to the following formula:

EP = GMP+ protein (sum of additional complementary amino acids/1,2).

The nutritional composition described above contains glycomacropeptide (GMP) as the primary protein source and additional complementary essential amino acids to complete the required protein profile. Additional complementary essential amino acids are supplied in a mixture containing arginine, cystine, histidine, tryptophan, tyrosine and leucine.

In the nutritional composition described above, the arginine content ranges from 33 to 50 milligrams of arginine/gram of protein equivalents and the leucine content of 47 to 76 milligrams of leucine/gram of protein equivalents. Of these, the amount of arginine added in addition to GMP-derived arginine ranges from 32 to 48.5 milligrams arginine/gram equivalent proteins, while the amount of leucine added in addition to GMP-derived leucine ranges from 39 to 65 milligrams leucine/ grams of equivalent proteins.

The applicant has discovered that leucine is an amino acid with a sensory profile with a very pronounced bitter taste. However, it is known to play an important role in nutrition as an essential amino acid. For these reasons, the dosage of leucine in the composition according to the present description, ranging from only 47 to 76 milligrams of leucine/gram of equivalent proteins, is greatly reduced compared to the state of the art, with minimal impact at the organoleptic level, in any case compensating for the GMP deficiency, while maintaining the role of an essential amino acid, as defined in the document Protein and amino acids requirements in human nutrition (WHO, Table 23 and Table 36).

The applicant has discovered that arginine is also a very bitter amino acid. Therefore, during the formulation development phase of the present specification, the Applicant performed experimental tests to compare the effects of different forms of arginine on the overall sensory profile of the GMP formulation in accordance with the present specification. The Applicant has found that it is preferable to use the arginine aspartate salt formulation over the pure arginine form because it is less bitter. Thus, in some embodiments, the quantitative contribution of arginine added to the formulation described herein, in addition to GMP-derived arginine, comes from arginine supplied in salt form, particularly arginine aspartate salt, helping to optimize the sensory profile of the product from a technological point of view.

This document notes that GMP inherently supplies reduced amounts of arginine, cystine, histidine and leucine, so in the composition described above, the total minimal portion of arginine, cystine, histidine and leucine comes from GMP, and the remaining majority comes from additional complementary essential amino acids , while in the composition described above, tryptophan and tyrosine come only from additional complementary essential amino acids.

In exemplary embodiments, which can be combined with all embodiments described herein, the nutritional composition has a tyrosine content of 112 to 188 milligrams of tyrosine/gram of equivalent proteins derived solely from additional amino acids and not from GMP.

According to exemplary embodiments, which may be combined with all embodiments described herein, in a nutritional composition, the total weight of the additional complementary essential amino acids is from 35% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids amino acids.

In further exemplary embodiments, the total weight of the additional essential complementary amino acids is from 45% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids.

In further exemplary embodiments, the total weight of the additional essential complementary amino acids is from 55% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains 12 to 22 milligrams of tryptophan/gram of equivalent proteins and 22 to 37 milligrams of histidine/gram of equivalent proteins. Of these, tryptophan is derived exclusively from accessory amino acids rather than GMP, while the amount of histidine added in addition to GMP-derived histidine ranges from 21 to 36.3 milligrams of histidine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains between 3.8 and 6.2 milligrams of cystine/gram equivalent protein. Of these, the amount of cystine added, in addition to cystine derived from GMP, ranges from 3.45 to 5.75

- 5,043,776 milligrams of cystine/gram of equivalent protein.

In additional embodiments, combined with all embodiments described herein, the nutritional composition described above, in addition to the amino acids arginine, cystine, histidine, leucine, tryptophan, and tyrosine mentioned above, also contains one, more, or all of the following amino acids: alanine, aspartic acid, glutamic acid, glutamine, glycine, isoleucine, lysine, methionine, proline, serine, threonine, valine. Of these, isoleucine, serine and valine originate only from GMP as they are present in it by nature, while alanine, aspartic acid, glutamic acid, glutamine, glycine, lysine, methionine, proline, threonine originate from both GMP and added to complete the nutritional profile. Thus, in possible embodiments, the nutritional composition described above may also include, in addition to GMP and the amino acids naturally present therein, 15 additional amino acids, namely, arginine, cystine, histidine, leucine, tryptophan, tyrosine, which are contained in GMP is low, as well as alanine, aspartic acid, glutamic acid, glutamine, glycine, lysine, methionine, proline and threonine.

The minimum phenylalanine content in the formulation described herein, in any case well below the minimum threshold for patients with PKU, comes exclusively from GMP where phenylalanine is naturally present. For example, GMP is used, which contains a maximum of 0.2 mg of phenylalanine per 100 grams of protein. In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains no more than 1.6, preferably 1.5, even more preferably 1.4 milligrams of phenylalanine/gram equivalent protein. For example, the phenylalanine content may range from 1.2 to 1.5, preferably 1.2 to 1.4 milligrams of phenylalanine/gram equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains between 51 and 75 milligrams of alanine/gram equivalent protein in the nutritional composition. Of these, the amount of alanine added, in addition to the GMP-derived alanine, ranges from 28 to 47 milligrams of alanine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition has an aspartic acid content of 86 to 130 milligrams aspartic acid/gram equivalent protein. Of these, the amount of aspartic acid added, in addition to the GMP-derived acid, ranges from 53 to 90 milligrams of aspartic acid/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition has a glutamic acid content of 78 to 99.5 milligrams glutamic acid/gram equivalent protein. Of these, the amount of glutamic acid added, in addition to the GMP-derived acid, ranges from 7 to 12 milligrams glutamic acid/gram equivalent proteins.

In exemplary embodiments, which can be combined with all embodiments described herein, the nutritional composition has a glutamine content of 47 to 79 milligrams of glutamine/gram of equivalent proteins derived solely from additional amino acids and not from GMP.

In exemplary embodiments, which can be combined with all embodiments described herein, the nutritional composition has a glycine content of from 58 to 96 milligrams of glycine/gram of equivalent protein. Of these, the amount of glycine added in addition to GMP-derived glycine ranges from 54 to 91 milligrams of glycine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains between 39 and 96 milligrams of isoleucine/gram equivalent protein in the nutritional composition. Of these, the amount of isoleucine added in addition to the GMP-derived isoleucine may range from 0 to 48 milligrams isoleucine/gram equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition has a lysine content of from 61 to 93 milligrams of lysine/gram of equivalent protein. Of these, the amount of lysine added in addition to GMP-derived lysine ranges from 39 to 65 milligrams of lysine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition has a methionine content of 15 to 23.5 milligrams of methionine/gram of equivalent protein. Of these, the amount of methionine added in addition to GMP-derived methionine ranges from 8 to 14 milligrams of methionine/gram of equivalent proteins.

- 6 043776

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains between 99 and 146.5 milligrams of proline/gram equivalent protein in the nutritional composition. Of these, the amount of proline added, in addition to the GMP-derived proline, ranges from 55 to 92.5 milligrams of proline/gram of equivalent proteins.

In exemplary embodiments, which can be combined with all embodiments described herein, the nutritional composition has a serine content of 28 to 69 milligrams serine/gram equivalent protein. Of these, the amount of serine added in addition to the GMP-derived serine may range from 0 to 34.5 milligrams of serine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains between 79 and 105.5 milligrams threonine/gram equivalent protein. Of these, the amount of threonine added, in addition to the GMP-derived threonine, ranges from 16 to 28 milligrams of serine/gram of equivalent proteins.

In exemplary embodiments, which may be combined with all embodiments described herein, the nutritional composition contains valine from 31 to 77 milligrams valine/gram equivalent protein. Of these, the amount of valine added, in addition to GMP-derived valine, can range from 0 to 38.5 milligrams of valine/gram of equivalent proteins.

Preferably, the aminogram which characterizes the formulations in accordance with the various embodiments has been developed and tested by the Applicant in such a way that, in addition to providing an adequate amount of complementary essential amino acids suitable for the purpose of dietary therapy for PKU, at the same time it ensures suitable sensory properties of the formulations themselves, so that they are also pleasant from an organoleptic point of view for those who take the compositions described above.

In accordance with possible embodiments that can be combined with all embodiments described herein, the nutritional composition described above is presented in powder form. A possible exemplary embodiment is a powder product containing a composition according to the present disclosure, which is to be taken after dissolution in a liquid, such as water or a similar suitable beverage. The powder product may include a mixture of dry ingredients, including the composition in accordance with the present description and, for example, carbohydrates, fats, vitamins, mineral salts, trace elements, and the like. The powder product may be partial or complete, depending on the recipe.

Other embodiments also relate to a liquid product or food beverage containing a composition in accordance with the embodiments described herein. A possible exemplary embodiment is a ready-to-drink liquid product or beverage that contains at least a formulation in accordance with embodiments and which may also contain, for example, vitamins and/or minerals and/or trace elements or other beneficial or functional substances. The liquid product may include a mixture of dry ingredients, including the composition in accordance with the present description, and liquid ingredients. Among the additional ingredients, dry or liquid, in addition to the composition in accordance with the present description, for example, carbohydrates, fats, vitamins, mineral salts, trace elements, etc. can also be provided in this case. The liquid product may be partial or complete, depending on the recipe.

According to possible embodiments, other food products which contain a composition according to the present description may also be provided, such as dairy products, in particular yoghurt, or confectionery products such as creams or puddings, bakery products such as bread, crackers, breadsticks, snacks, cookies, cakes, sweet snacks, confectionery and pasta or the like, or other compositions of varying density or viscosity. Also, other nutritional beverages may be provided that utilize the nutritional composition of the present disclosure.

Thus, the provision that the composition in accordance with the embodiments described herein can be the skeleton or common basis for various food products or beverages that can be manufactured falls within the spirit of the present description. This backbone or common base may consist primarily of the GMP protein portion and additional complementary essential amino acids and therefore provides the basis for the various recipes of the different foods that may be prepared.

In addition, the resulting products formulated according to the embodiments described herein may contain carbohydrates and fats added thereto in varying amounts.

The products thus obtained may be complete or incomplete products. If you yourself

- 7 043776 the product contains a certain amount of proteins, carbohydrates and fats sufficient to meet the nutritional needs of a patient with phenylketonuria, it is considered complete, otherwise it is defined as incomplete.

In accordance with possible embodiments, which can be combined with all the embodiments described herein, carbohydrates and fats may also be present in food products that include the nutritional composition described above.

In these foods, carbohydrates can provide 20% to 50% of the food's energy, and fats can provide 1% to 50% of the food's energy.

With regard to carbohydrates, in possible embodiments, the use of monosaccharides in the resulting food product recipes is preferably limited, as much as possible, in favor of the use of polysaccharides, such as, for example, starches, maltodextrin, trehalose and lactose.

In possible embodiments, in particular, the composition may contain trehalose in an amount of from 35% to 95% by weight, based on the total weight of carbohydrates present in the food product that includes the nutritional composition as described herein.

Specifically, trehalose is a disaccharide consisting of two glucose molecules joined by an α-α(1,1)' glycosidic bond. Its chemical structure makes it particularly stable from a kinetic and thermodynamic point of view, and makes it a non-reducing sugar, so not very prone to react with other substances within food matrices. In addition to this, trehalose has the ability to protect proteins and their structure in certain stressful situations.

In addition, trehalose is involved in modulating salty and bitter tastes in food matrices. In the composition according to the present description, the use of trehalose advantageously allows the masking of bitter and salty tastes resulting not only from certain amino acids, but also from certain mineral salts. Thus, the beneficial use of trehalose as a primary carbohydrate source also plays a role in reducing the off-flavors typical of these formulations.

In products prepared herein, added carbohydrates may provide from about 20% to about 50% of the energy.

According to exemplary embodiments, which may be combined with all embodiments described herein, the weight ratio between proteins and carbohydrates in a food product that includes the nutritional composition herein may range from 0.85 to 2.5 . Possible examples include 0.85, 0.9, 0.95, 1, 1.25, 1.3, 1.5, 1.75, 2, 2.25, 2.5. These values, both in terms of range and in terms of specific values, can be selected for both powder and liquid products, according to needs.

According to exemplary embodiments, which may be combined with all embodiments described herein, the weight ratio between proteins and fats in a food product that includes the nutritional composition herein may range from 10 to 130. Possible examples may include: be 10, 12, 18, 25, 30, 35, 40, 45, 50, 60, 70, 80, 85, 90, 100, 110, 120, 130. In particular, in the case of a powder product, the mass ratio between proteins and fats in a food product may be, for example, from 60 to 130. In the case of a liquid product, the weight ratio between proteins and fats in the food product may be, for example, from 10 to 90.

With regard to fats, fats may be present in food recipes prepared according to the present disclosure, such as, for example, polyunsaturated essential fatty acids, in particular omega-3 fatty acids, e.g. alpha-linolenic acid, eicosapentaenoic acid (EPA) ), docosahexaenoic acid (DHA), and omega-6s, such as linoleic acid and arachidonic acid (ARA), to enrich the nutritional profile of foods. For example, ARA, EPA and DHA may be used in recipes.

Additionally, other vegetable fats, particularly vegetable oils, can also be included in recipes to enrich the nutritional profile of foods.

Possible examples are canola oil, sunflower oil, palm/palm kernel oil, soybean oil, flaxseed oil, safflower oil, coconut oil and shea butter.

In addition, the addition of medium chain fatty acids (medium chain triglyceride, MCT) is also possible.

In products prepared herein, added fats may provide from about 1% to about 50% of the energy.

In addition, in food recipes prepared in accordance with the present description, plant fibers may be present in amounts up to 10% by weight of the recipe.

The resulting foods may also contain minerals, trace elements and vitamins to achieve a balanced micronutrient profile.

In addition, other embodiments described herein relate to a method of making a nutritional composition for constructing a diet for phenylketonuria. In accordance with

- 8 043776 In one embodiment, the method includes mixing a glycomacropeptide (GMP) as the main source of proteins and additional complementary essential amino acids to complete the desired protein profile, supplied in a mixture containing arginine, cystine, histidine, tryptophan, tyrosine and leucine. Arginine and leucine are supplied in such quantities that the nutritional composition contains 33 to 50 milligrams of arginine/gram of equivalent protein and 47 to 76 milligram of leucine/gram of equivalent protein.

In additional embodiments, a method of making food products in powder or liquid form using a composition as described herein may also be provided.

If the product is formulated as a dry powder, the method may involve mixing the dry ingredients, including the composition described above, in suitable mixers. The product is then dispensed and packaged in suitable multi-serve containers, such as cans or jars, or single-serve containers, preferably in a modified atmosphere.

When preparing a ready-to-use liquid product, the method may involve mixing a selected liquid, such as water, dry powder ingredients, including the composition described herein, and liquid ingredients. The resulting liquid mixture is stirred for a suitable time, after which it is microbiologically stabilized by heat treatment (UHT or pasteurization) and packaged in packages of various sizes, multi-serving or single-serving. Alternatively, the mixture may be stabilized by cold pasteurization with high pressure processing (HHP, High Hydrostatic Pressure or HPP, High Pressure Processing); in this case, depending on the protein content, a final product with altered density or viscosity can be obtained.

Thus, aspects of the various products obtained using the composition and method in accordance with the present description, taken individually or in combination with each other in accordance with the various embodiments described herein, may be the following:

very low phenylalanine content, not exceeding 1.4 milligrams of phenylalanine/gram of equivalent proteins;

the protein part, consisting exclusively of GMP and additional essential complementary amino acids;

an aminogram in which, relative to the total amino acids, about 35% to about 65% comes from GMP and the remaining about 65% to about 35% comes from free amino acids. For example, about 40% comes from GMP, and the remaining about 60% comes from free amino acids;

a complete profile of all amino acids, both essential and non-essential;

aminogram, which allows the supply of complementary essential amino acids in quantities suitable for the purpose of dietary therapy for PKU, while obtaining suitable sensory properties of the compositions, in such a way that they are also pleasant from an organoleptic point of view for those who take the above-described compositions, as described below with reference to experimental tests carried out by the Applicant;

high tyrosine levels, 112 to 188 milligrams of tyrosine/gram of equivalent protein, consistent with the nutritional needs of patients with PKU, given that in PKU, tyrosine becomes an essential amino acid;

Due to the special composition of amino acids, there is an adequate amount of large neutral amino acids (LNAA), which can compete with the supply of phenylalanine at the brain level. For example, in some embodiments, from about 0.360 to about 0.625 grams LNAA/gram equivalent proteins may be present. A possible example is approximately 0.448 grams LNAA/gram equivalent proteins.

The Applicant has found that among the LNAAs, leucine has the worst organoleptic profile and a very pronounced bitter taste. However, the Applicant also noted that it also plays an important role in nutrition as it is an essential amino acid. For these reasons, the dosage of leucine in the formulation as disclosed herein, generally ranging from 47 to 76 milligrams of leucine/gram of equivalent protein, is significantly reduced compared to the state of the art, meaning it has minimal impact at the sensory level and in any case corrects the deficiency GMP, maintaining the role of an essential amino acid.

In addition, the following features further characterize products obtained using embodiments of the composition described herein:

in the case of nutritionally incomplete products, they may have a reduced calorie content due to the reduced content of carbohydrates and, above all, simple sugars;

foods can be high in micronutrients usually missing in patients with PKU (such as, for example, calcium, zinc, vitamin B12, selenium, iron), without exceeding

- 9 043776 large dose in order to meet higher levels of established intake;

where possible, ingredients such as gluco-oligosaccharides/fructo-oligosaccharides GOS/FOS, prebiotic compounds such as inulin and chokeberry (antioxidant) are added, which, depending on the target patients, can improve the nutritional profile of the products in relation to PKU.

Examples

Example 1.

In the following tables. 1 and 2 show the ranges of GMP values and free amino acids present in the formulations in accordance with exemplary embodiments, which may be suitable for the manufacture of various types of products, for example, as described above. Values in table. 1 and 2 are expressed as milligrams per gram equivalent proteins.

In table 1 and 2, taken as a whole, provide examples in which the total weight of the additional complementary essential amino acids is from 45% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids.

The GMP used contains 0.2 mg phenylalanine per 100 grams of equivalent proteins, and the established upper limit for phenylalanine is 1.4 mg/gram of equivalent proteins.

In particular, in table. 1 below, reference is made to exemplary embodiments in which the total weight of the additional essential complementary amino acids is from 55% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids.

Table 1

Amino acid Unit Number of amino acids from GMP Number of free acids added General aminogram Alanin milligram (mg) 22.86-27.94 28.2-47 51.06-74.94 Arginine milligram (mg) 1.08-1.32 32.85-48.18 33.93-49.5 Aspartic acid_______ milligram (mg) 32.58-39.82 53.85-89.75 86.43-129.57 Cystine milligram (mg) 0.35-0.43 3.45-5.75 3.8-6.18 Glutamic acid_______ milligram (mg) 71.37-87.23 7.2-12 78.57-99.23 Glutamine milligram (mg) - 47.25-78.75 47.25-78.75 Glycine milligram (mg) 4.14-5.06 54.3-90.5 58.44-95.56 G istidin milligram (mg) milligram (mg) 0.36-0.44 21.75-36.25 22.11-36.69 Isoleucine 39.15-47.85 0-47.85 39.15-95.7 Leucine milligram (mg) 8.28-10.12 39-65 47.28-75.12 Lysine milligram (mg) 22.86-27.94 39-65 61.86-92.94 Methionine milligram (mg) 7.65-9.35 8.25-13.75 15.9-23.1 Proline milligram (mg) 44.37-54.23 55.28-92.13 99.65-146.36 Serin milligram (mg) 28.08-34.32 0-34.32 28.08-68.64 Threonine milligram (mg) 63.45-77.55 16.5-27.5 79.95-105.05 Tryptophan milligram (mg) - 12.75-21.25 12.75-21.25 Tyrosine milligram ( ^mg ) - 112.5-187.5 112.5-187.5 Valin milligram (mg) 31.5-38.5 0-38.5 31.5-77 Phenylalanine milligram (mg) 1.26-1.54 - 1.26-1.54

In the table below. 2 specifically refers to exemplary embodiments in which the total weight of the additional complementary essential amino acids is from 45% to 55% of the total weight represented by the weight of the GMP proteins together with the weight of the additional complementary essential amino acids.

- 10 043776

table 2

Amino acid Unit Number of amino acids from GMP Number of free amino acids added General aminogram Alanin milligram (mg) 28.42-36.92 22.75-37.91 51.17-74.83 Arginine milligram (mg) 1.29-1.68 32.64-47.87 33.93-49.54 Aspartic acid milligram (mg) 40.48-52.58 46.10-76.84 86.58-129.42 Cystine milligram (mg) 0.43-0.56 3.38-5.63 3.81-6.19 Glutamic acid milligram (mg) 88.71-115.23 10.09-16.82 98.81-132.05 Glutamine milligram (mg) - 47.29-78.82 47.29-78.82 Glycine milligram (mg) 5.17-6.71 53.30-88.83 58.46-95.54 G istidin milligram (mg) milligram (mg) 0.43-0.56 22.23-35.63 22.66-36.19 Isoleucine 48.66-63.21 0-63.20 48.66-126.41 Leucine milligram (mg) 10.34-13.42 36.84-61.4 47.18-74.82 Lysine milligram ( ^mg ) 28.42-36.92 33.25-55.41 61.67-92.33 Methionine milligram ( ^mg ) 9.47-12.31 6.08-10.14 15.56-22.44 Proline milligram (mg) 55.12-71.60 4473.74.55 99.85-146.15 Serin milligram (mg) 34.88-45.31 0-45.31 34.88-90.61 Threonine milligram (mg) 78.81-102.36 1.06-1.77 79.87-104.13 Tryptophan milligram (mg) - 12.75-21.25 12.75-21.25 Tyrosine milligram ( ^mg ) - 112.5-187.5 112.5-187.5 Valin milligram (mg) 39.19-50.90 0-50.90 39.19-101.80 Phenylalanine milligram (mg) 1.57-2.03 - 1.57-2.03

In the next table. 3, alternatively, shows the GMP values and free amino acids present in a particular illustrative composition in accordance with the present description. Values in table. 3 are expressed as milligrams per gram equivalent proteins.

- 11 043776

Table 3

Amino acid Unit Amount of amino acid from GMP Number of free acids added General aminogram Alanin milligram (mg) 25.4 37.6 63 Arginine milligram (mg) 1.2 43.8 45 Aspartic acid milligram (mg) 36.2 71.8 108 Cystine milligram (mg) 0.39 4.6 5 Glutamic acid milligram (mg) 79.3 9.6 89 Glutamine milligram (mg) - 63 63 Glycine milligram (mg) 4.6 72.4 77 G istidin milligram (mg) 0.4 29 29 Isoleucine milligram (mg) 43.5 - 44 Leucine milligram (mg) 9.2 52 61 Lysine milligram (mg) 25.4 52 77 Methionine milligram (mg) 8.5 AND 19 Proline milligram (mg) 49.3 73.7 123 Serin milligram (mg) 31.2 - 31 Threonine milligram (mg) 70.5 22 92 Tryptophan milligram (mg) - 17 17 Tyrosine milligram (mg) - 150 150 Valin milligram (mg) 35 - 35 Phenylalanine milligram (mg) 1.4 - 1.4

Example 2.

In table 4 below shows an example of a powdered food product containing a nutritional composition in accordance with the present description and containing GMP, for consumption after reconstitution with water. This example is an incomplete product that provides approximately 300 kcal/100 g and is characterized by the following macronutrient profile, presented as a range of amounts in grams per 100 g of powdered food product.

Table 4 g/100 g

Proteins_______________30-45________

Carbohydrates___________20-35________

Fats______________0.35-2_______

This example of a powdered food product according to the present disclosure may consist of the following ingredients, expressed as amounts in grams per 100 g of powdered food product (Table 5).

- 12 043776

Table 5

g/100 g Alanin 1.6 Cystine 0.2 Glutamine 2.8 Glycine 3.2 G istidin 1.3 Leucine 2.3 Methionine 0.5 Proline z, z Threonine 1 Tryptophan 0.8 Tyrosine 6.6 Arginine aspartate 3.4 Lysine aspartate 3.6 Lysine glutamate 1 GMP 22 Trehalose 20 G.O.S. 1.7 F.O.S. 0.2 DHA 2.6 Potassium carbonate 1.5 Calcium phosphate 4.6 Potassium phosphate 1.7 Magnesium carbonate 1.2 Ferrous sulfate 1.6 Zinc sulfate 0.04 Copper sulfate 0.003 Manganese sulfate 0.005 Sodium selenite 0.0002 Chromium chloride 0.0003 Sodium molybdate 0.0002 Potassium iodide 0.0004 Carnitine 0.04 Kholin 1 Inositol 0.13 Vitamins 0.7 Sucralose 0.04 Aromatic compounds 12

Example 3.

An example of a liquid food product containing a nutritional composition in accordance with the present description and containing GMP, ready for consumption, is shown below in table. 6. This example is a partial product that provides approximately 60 kcal/100 g and has the following macronutrient profile presented as a range of grams per 100 g of liquid food product.

Table 6

g/100 g Squirrels 6-9 Carbohydrates 4-6 Fats 0.1-1

This example liquid product according to the present disclosure may consist of the following ingredients, presented as amounts in grams per 100 g of powdered food product (Table 7).

Table 7

g/100 g Alanin 0.3 Cystine 0.035 Glutamine 0.5 Glycine 0.55 G istidine 0.2 Leucine 0.4 Methionine 0.08 Proline 0.6 Threonine 0.15 Tryptophan 0.13 Tyrosine 1.13 Arginine aspartate 0.6 Lysine aspartate 0.61 Lysine glutamate 0.15 GMP 3.8

- 13 043776

Trehalose 2.8 Starch 2.1 Inulin 2 DHA 0.15 Sodium citrate 0.4 Potassium phosphate 0.23 Calcium phosphate 0.7 Magnesium carbonate 0.2 Potassium chloride o, s Ferrous sulfate 0.025 Zinc sulfate 0.006 Copper sulfate 0.0006 Magnesium sulfate 0.0007 Sodium Selenite 0.00004 Chromium chloride 0.00004 Sodium molybdate 0.00003 Potassium iodide 0.00006 Carnitine 0.006 Kholin 0.2 Inositol 0.02 Vitamins 0.1 Aromatic compounds 0.7 Dyes 0.01 Aronia chokeberry 0.004 Sucralose 0.01 Sunflower lecithin 0.4 Water 80.4

Experimental tests.

The composition according to the present description is complex and provides the presence of various ingredients such as GMP, free amino acids, as well as possible carbohydrates, minerals and trace elements. It follows that the final taste of the composition depends on the taste of the individual ingredients used, on their interaction in the matrix and possibly on the technological processes to which the food product is subjected during production. The composition in accordance with the present description was developed and experimentally studied by the Applicant with the aim of optimizing each of these aspects and ensuring maximum stability of the products throughout their shelf life.

As evidence of these organoleptic improvements, the Applicant conducted a sensory evaluation of the powder formulation in accordance with the examples presented in Table. 3 and 4, compared to two other GMP products (Product 1 and Product 2).

In the next table. 8 summarizes the characteristics of an exemplary powder formulation in accordance with this disclosure and comparison products 1 and 2.

Table 8

Product Features Composition according to an example of the present invention Product 1 Product 2 Energy kcal/10 g EP 75 kcal 60 kcal 106 kcal Energy from proteins (%) 54% 66% 38% Energy from carbohydrates(%) 40% 21% 57% Energy from fat (%) 5% 13% 25% g proteins/100 g 43 g 56 g 31 g g carbohydrates/100 g 32 g 18 g 47 g g fat/100 g 1.7 g 4.7 g 9.2 g

Organoleptic analysis was carried out by a trained group of 9 tasters. The parameters were assessed on the following scale: Intensity parameter Significance

Imperceptible

Slightly noticeable

Moderately perceptible

Clearly perceptible

Very noticeable

Extremely noticeable

As shown in table. 9 below, the sensory analysis performed showed that the intensity of bitter and sour tastes is much lower in the powder formulation according to the example of the present invention than in the two comparison products 1 and 2.

- 14 043776

Table 9

Composition according to the example herein Product 1 Product 2 Bitter taste 1D 2.3 3.2 Sour taste 1.3 1.7 1.9

It is understood that modifications and/or additions of parts and/or steps can be made to the nutritional composition for constructing a diet for phenylketonuria and the corresponding production method described hereinbefore without departing from the scope and scope of the present invention.

It is also apparent that while the present invention has been described with reference to certain specific examples, one skilled in the art would certainly be able to develop many other equivalent forms of the PKU diet nutritional composition and corresponding method of manufacture having the characteristics set forth in the claims, and , therefore, everything included in the thereby defined area of protection.

Claims (16)

CLAIM 1. A nutritional composition suitable for constructing a diet for phenylketonuria, containing glycomacropeptide (GMP) as the main protein source and additional complementary essential amino acids to complete the required protein profile, supplied in a mixture containing arginine, cystine, histidine, tryptophan, tyrosine and leucine, the content of arginine in the specified nutritional composition ranges from 33 to 50 milligrams of arginine/gram of equivalent proteins and leucine from 47 to 76 milligrams of leucine/gram of equivalent proteins. 2. The nutritional composition of claim 1, wherein the tyrosine content ranges from 112 to 188 milligrams of tyrosine/gram of equivalent protein. 3. The nutritional composition of claim 1 or 2, wherein the total weight of said additional complementary essential amino acids is from 35% to 65% of the total weight represented by the weight of the GMP proteins together with the weight of said additional complementary essential amino acids. 4. Nutritional composition according to claims 1, 2 or 3, in which the tryptophan content is from 12 to 22 milligrams of tryptophan/gram of equivalent proteins, histidine from 22 to 37 milligrams of histidine/gram of equivalent proteins and cystine from 3.8 to 6.2 milligram cystine/gram equivalent protein. 5. The nutritional composition according to any one of claims 1 to 4, wherein said arginine supplied as a complementary essential amino acid, in addition to GMP-derived arginine, is derived from arginine supplied in the form of an arginine salt, in particular an arginine aspartate salt. 6. A food product containing a composition according to any one of claims 1 to 5. 7. A food product according to claim 6, also containing carbohydrates and fats, wherein carbohydrates provide from 20% to 50% of the energy of the specified product, and fats supply from 1% to 50% of the energy of the specified product. 8. A food product according to claim 6 or 7, wherein the weight ratio between proteins and carbohydrates in said product, which includes said nutritional composition, is in the range of 0.85 to 2.5, and the weight ratio between proteins and fats in said product , which includes the specified nutritional composition, ranges from 10 to 130. 9. A food product according to any one of claims 6, 7 or 8, wherein the carbohydrates comprise trehalose and the trehalose content is from 35% to 95% by weight, based on the amount of total carbohydrate present in the product. 10. A drink containing the composition according to any one of claims 1-5. 11. The beverage of claim 10, further comprising carbohydrates and fats, wherein carbohydrates provide 20% to 50% of the energy of said beverage and fats provide 1% to 50% of the energy of said beverage. 12. The beverage according to claim 10 or 11, wherein the weight ratio between proteins and carbohydrates in said beverage, which includes said nutritional composition, is in the range of 0.85 to 2.5, and the weight ratio between proteins and fats in said beverage, which includes the specified nutritional composition, ranges from 10 to 130. 13. The beverage according to any one of claims 10, 11 or 12, wherein the carbohydrate comprises trehalose and the trehalose content is from 35% to 95% by weight, based on the amount of total carbohydrate present in the beverage. 14. A method of preparing a nutritional composition suitable for constructing a diet for phenylketonuria, the method comprising mixing a glycomacropeptide (GMP) as the main source of proteins and additional complementary essential amino acids to complete the desired protein profile, supplied in a mixture containing arginine, cystine, histidine , tryptophan, tyrosine and leucine, wherein arginine and leucine are supplied in such quantities that in the specified nutritional composition the arginine content is from 33 to 50 milligrams of arginine/gram of equivalent proteins, and of leucine from 47 to 76 milligrams of leucine/gram of equivalent proteins. - 15 043776 15. A method for producing a food product suitable for constructing a diet for phenylketonuria, wherein said method provides for the presence and mixing together of at least: a nutritional composition according to any one of claims 1 to 5, carbohydrates and fats. 16. A method for producing a drink suitable for building a diet for phenylketonuria, wherein said method ensures the presence and mixing together of at least: a nutritional composition according to any one of claims. 1-5, carbohydrates and fats.