Infant formula with improved fat composition

DESCRIPTION

INFANT FORMULA WITH IMPROVED PROTEIN CONTENT

The present invention relates to an improved infant formula which at least contains an easily digestible lipid component, a protein component, and a prebiotic component.

In infants, particularly in infants younger than 6 months, the digestive system has to develop and adapt to food. Therefore, in the first months of life, infants are usually fed special infant formulas.

Usually these infant formulas are well tolerated. However, in a limited number of cases, conventional formulas may give minor problems, in particular with regard to processes occurring in the gastrointestinal tract.

More particularly, in the research leading up to the present invention, it was recognized by the inventors that conventional formulas could cause undesirable changes in the natural intestinal flora and / or artificial intestinal flora. They also recognized that conventional infant formulas may provide the wrong type and / or incorrect amounts of substrate; and / or may have an incorrect composition of dietary components.

These defects in conventional formulas may result in unwanted processes in the gastrointestinal tract and / or an undesired change in natural processes. These defects may give rise to symptoms such as:

- an undesirable build up of stools (eg diarrhea, constipation or an unusual color);

1/38 high local gas production, which can result in spasms in the infant's intestines and / or dilation and delayed stomach emptying;

- a decrease in the bioavailability of bivalent cations;

unwanted levels of aggressive agents in faeces, in particular agents that may irritate and / or damage epithelial cells (such as those of the gastrointestinal tract lining and / or skin) that include but are not limited to proteases.

As a result, infants who are fed conventional formulas may suffer from lack of appetite and / or regurgitation of food (frequent in the case of bowel disorders). There may also be changes in the natural process of stomach emptying and an irregular pattern of consumption that can lead to hunger, whereby infants may precipitously consume large volumes of the formula, which may lead to aerophagia and / or regurgitation. Excessive.

Also, as a result of the above problems, causes and / or factors, infants who are fed conventional formulas may suffer from impaired immune function that may be observed in the rapid development on the skin of the buttocks of a rash; malnutrition (poor growth), dysfunction (colic) and even bowel damage (intolerance to food components).

Genzel-Boroviczény et al (Eur. J. Pediatr. (1997) 156: 142-147) report on the composition of fatty acids in human milk during the first month after term and preterm birth; comprises, inter alia, oleic, linoleic, α-linolenic, arachidonic and docosahexaenoic acid. Obermeier et al. (Isotopes. Environ. Health Stud. (1999), 35: 119-125) report that galactose applied orally to the mother is incorporated into milk,

2/38 especially in lactose and neutral oligosaccharides, which are relevant to the infant's defenses against bacterial and viral adhesion.

Thus, there is a need for a formula for infants that has the improved characteristics for preventing and / or alleviating one or more, and preferably all, of the above problems. The present invention provides this type infant formula.

In this regard, it is important that the formula of the invention be capable of simultaneously preventing, alleviating and / or reducing all of the above problems. This is because, particularly for parents, it is difficult to establish the specific cause of the problem (s). Thus, by using the infant formula of the invention which can prevent and / or alleviate all these problems simultaneously, it is assured to parents that by using the infant formula of the invention, all these problems can be reduced, regardless of the cause that causes them. originate, and without having to switch from one formula to another until you find an infant formula that reduces the problem (s) that are currently being produced and without causing any of the problem (s) mentioned above.

Usually, when one or more of the above problems occur / persist, parents usually switch to another formula or if this does not help - consult a pediatrician. However, of all commonly used infant formulas, none have features that can simultaneously prevent or alleviate all of the above problems. Thus, switching to another formula may not reduce the problem and / or may give rise to one or more of the above problems. Also, the infant may have trouble getting used to the (taste of) the new formula, with which one may lose one's appetite. The latter may be a problem.

Particularly when - due to the aforementioned problems - the formula is or has to be changed several times.

In the research leading up to the present invention, the inventors have confirmed that some of the main causes of the alteration (s) of processes in the gastrointestinal tract that may occur with conventional infant formulas include:

- the fact that, in triglycerides present in (the lipid component of) the conventional infant formula, undesirable large amounts of palmitic acid, myristic acid, and / or stearic acid are present in the glycerin Sn1 or Sn3 position;

- the fact that conventional infant formulas contain large undesirable amounts of bivalent cations, and in particular large undesirable amounts of calcium ions. In particular, this is a problem when at the same time large amounts of palmitic acid, myristic acid, and / or stearic acid are present at the Sn1 or Sn3 position of the present triglycerides, that is, as mentioned above;

the fact that conventional infant formulas contain undesirable large amounts of substrates suitable for microorganisms, i.e. for intestinal flora of natural origin.

Thus, according to the invention, the above mentioned problems are solved with an infant formula according to the claims.

Also, as mentioned below, preferably the amount of phosphorus in the infant formula should be as low as possible, as for example as defined below. Inter alia according to the invention, these are obtained using proteins, and in particular

In particular protein hydrolysates, with a low phosphorus content, also as described below.

In this regard, the use of a protein hydrolyzate also ensures that the protein components present in the infant formula of the invention do not sensitize infants. The use of hydrolysates also ensures that protein components can be easily absorbed into the gastrointestinal tract, and thus not interfere (too much) with the processes that take place in the intestines (and in particular the colon).

Also, as will be further described below, the use of a protein hydrolyzate - together with the favorable osmotic value provided by the formula of the invention - will stimulate the natural processes of stomach emptying, which prevent and / or reduce regurgitation.

A formula for infants is provided that contains a sufficient amount of substrate for the creation and maintenance of beneficial flora throughout the gut, especially in the colon. This ensures uniform production of gases and beneficial substances such as short chain fatty acids, and also prevents the release of excessive amounts of aggressive agents. In humans, lactose and oligosaccharides in breast milk provide this substrate. In the conventional infant formula, lactose is the only source. More preferably, it has been found that in the general formula made according to the invention there is a source of oligosaccharides, for example in addition to or in place of lactose, the latter especially for infants who are (partially) lactose intolerant.

Thus, the present invention provides an infant formula comprising the following components:

(a) at least one protein component (hereinafter also referred to as component A);

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b) at least one lipid component that can easily be digested by an infant (component B);

c) at least one prebiotic component (component C), comprising a mixture of one or more transgalactooligosaccharides and one or more fructooligosaccharides;

e) long chain polyunsaturated fatty acids linoleic acid, α-linolenic acid, oleic acid, arachidonic acid and docosahexaenoic acid;

optionally at least one viscosity enhancing component (component D);

and optionally also containing any component of the infant formula known per se, including but not limited to those described below (which will be referred to as additional components); and wherein the components A through D and one or more additional components are preferably as described below.

The protein component may comprise whole proteins and / or a protein hydrolyzate, or a mixture thereof, and is most preferably low in phosphorus, such as a phosphorus content of less than 0.75 g / 100 g protein. Also, when whole proteins are present, they are preferred because they are easily digested by the infant, as for example described below.

Preferably, the entire infant formula of the invention has a phosphorus content as defined below.

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As a source of the protein component, mammalian milk is preferably used, for example bovine milk, goat milk or equine milk.

In particular, an infant formula of the invention may be characterized by:

the protein component a) is a protein hydrolyzate obtained by hydrolyzing a precursor material comprising a milk protein, a mixture of milk proteins, and / or a milk protein preparation with a combination of at least one endo and at least one exoproteinase;

and may additionally be characterized by:

at least one lipid component b) comprising at least one fatty acid triglyceride and / or a mixture of fatty acid triglycerides, wherein:

- palmitic acid residues represent more than 10%, up to for example 30% or more, preferably 16 - 24%, of all fatty acid residues present in triglycerides; and wherein the triglycerides wherein the palmitate residue is in the Sn1 or Sn3 position is not greater than 16%, more preferably not greater than 13, and most preferably not greater than 10.7% of all triglycerides present.

More preferably, an infant formula of the invention which for example contains two or more, and preferably three or more of the components A through D as mentioned above - is further characterized in that said formula for the most part, and preferably essentially, contains ions. of calcium that have a high biological availability, as for example described below.

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Preferably, the infant formula of the invention is a powder which, before use, is dissolved by the end user (i.e. the parents) in hot or cold water, and preferably warm, or other suitable liquid support, for example in a amount of 9 - 17 g of powder per 100 ml of water. This powder, optionally packaged in a suitable container such as a can, box, bag, vial or sachet, as well as the ready-to-use formula obtained by dissolving the powder, constitutes some aspects of the present invention.

In a more preferred infant formula containing at least components A, B and C, the relative amounts of the above components will generally be as follows:

- component A: 6.4 to 16.0% by weight

- component B: 18 to 29% by weight

- component C: 3.0 to 15% by weight

- component D: 0 to 15% by weight

Additional components: 0 to 60% by weight up to a total of 100% by weight of the final formula.

In the infant formula containing the four components A, B, C and D, the relative amounts of the above components are generally as follows:

- component A: 6.4 to 16.0% by weight

- component B: 18 to 29% by weight

- component C: 3.0 to 15% by weight

8/38 component D: 1.5 to 15 wt%

Additional components: 0 to 60% by weight up to a total of 100% by weight of the final formula.

It is contemplated that various infant formulas of the invention may be developed, with for example different properties, a variety of flavors, and / or for different ages of the infant.

In particular, it is contemplated that infant formulas of the invention for infants under 6 months may be developed, as well as infant formulas of the invention for infants over 4 months, and in particular over 6 months of age. age. These formulas will be further discussed below.

Proteins which are preferably used as component A have a relatively low phosphorus content. 100 g of protein should provide less than 0.75 g of phosphorus, preferably less than 0.72 g and even more preferably less than 0.69 g. Whey proteins, dephosphorylated or soy casein protein and their derived mixtures, as well as casein-specific protein fractions - such as those from which part of α-caseins has been deleted, and preferably most of them are suitable sources. .

Protein hydrolysates which are preferably used as component A constitute a further separate aspect of the invention, as well as the method for preparing said preferred hydrolysates. It is contemplated that these hydrolysates may serve other applications than their use in the formulas of the invention as further discussed below.

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Preferred protein hydrolysates A - when used in the amounts indicated below - generally provide the final formula of the invention with one or more - and preferably all of the following properties:

- a peptide profile - that is for hydrolysate-derived proteins - as described below;

good taste, and in particular a non-bitter taste, is generally an extremely important feature for infant formulas, and in particular for infant formulas of the invention;

a free amino acid content of less than 10 g, and preferably less than 7 g per 100 g of protein equivalent (calculated by multiplying Kjeldahl nitrogen by 6.25; where the amount of nitrogen that is measured is determined by the amount of proteins, peptides, amino acids and other sources of nitrogen in the product) based on the final formula;

an osmolality of less than 270 mOsmol / 1, preferably less than 240 mOsmol / 1, more preferably less than 230 mOsmol / 1. Inter alia, an osmotic value in this range will ensure that the infant formula of the invention does not alter (e.g. inhibit) the natural process of emptying the infant's stomach;

- low sensitization, as for example determined by the tests described by JJ Pahud & K. Swartz in Proc. of the Tropical Conference, Virginia 1985, AOAC, 264-271; and / or Piacentini et al., Allergy 1994, 48, 361-364 or any other suitable test.

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In the tests described by Pahud & Swartz or Piacentini, the infant formula of the invention preferably shows a degree of sensitizing properties which is preferably essentially zero.

Additionally, according to a preferred aspect, when the infant formula of the invention contains the preferred lipid component B described below, compared to conventional infant formulas, the formula of the invention may have a reduced overall fat content. According to this aspect of the invention, the infant formula of the invention may have an overall fat content (expressed as total lipid content) of less than 46 wt%; compared to 48 wt.% or more for conventional infant formulas.

In the formula of the invention, medium chain triglycerides (MCTs) may be used to partially replace triglycerides containing palmitic acid residues. In this case, it is preferred that these MCTs be used in an amount of less than 20% by weight of the total lipid component B.

Further, as further discussed below, when the infant formula of the invention contains the preferred lipid component B, an infant formula of the invention may not require that calcium amounts be present as high as the calcium content required in conventional formulas.

In particular, according to this aspect of the invention, an infant formula of the invention for infants younger than 6 months may have a calcium content (expressed in mg of calcium per 100 kcal of energy content of the formula) of less than 80 mg. 100 kcal, and preferably less than 77 mg / 100 kcal (e.g. 70 mg / 100 kcal); compared with 82 - 120 mg / 100 kcal of conventional infant formulas. To provide the infant with sufficient calcium, the calcium concentration should be greater than 50 mg / 100kcal.

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An infant formula of the invention for infants older than 6 months may have a calcium content (expressed in mg of calcium per 100 kcal of energy content of the formula) of less than 122 mg / 100kcal, and preferably less than 116 mg / kg. 100 kcal (e.g. 110 mg / 100 kcal); compared to 125 mg / 100kcal or more for conventional infant formulas.

This reduction in calcium content may have certain advantages, such as reduced formation of fatty acid soaps in the intestine and increased natural exposure of calcium ions to enterocytes in the duodenum and other parts of the intestine and intestinal flora. Additionally it has been found that fewer technological problems related to sedimentation have occurred, for example during the preparation of the formula.

Calcium is preferably added to the infant formula of the invention as soluble (soluble) salts, such as hydroxide or chloride salts. The tribasic phosphate and citrate salts are insoluble. Additionally, it was intended to prevent insoluble calcium salts such as phosphates and / or citrates from forming during production.

Also, as in the infant formula, generally the amount of phosphorus present depends on the calcium content, where generally the calcium-phosphorus ratio in conventional infant formulas is generally between 1.5 and 2.0, and usually approximately 2, the infant formula. of the invention may further have a reduced phosphorus content.

For example, based on the above-mentioned calcium content and a general calcium-phosphorus ratio of approximately 2, an infant formula of the invention for infants under 6 months may have a phosphorus content (expressed in mg phosphorus per 100 energy content of the formula) less than 40 kcal

12/38 mg / 100 kcal, and preferably less than 38.5 mg / 100 kcal (e.g. 38 mg / 100 kcal); whereas an infant formula of the invention for infants over 6 months of age may have a phosphorus content (expressed in mg of phosphorus per 100 kcal energy content of the formula) of less than 61 mg / 100kcal, and preferably less than 58%. mg / 100 kcal (e.g. 38 mg / 10 Okcal).

This reduced phosphorus content may have some advantages, such as lower sedimentation during manufacture and increased bioavailability of bivalent cations.

Where in addition to phosphorus provided by proteins and / or phospholipids, extra phosphorus must be added, this addition is especially effected by the addition of monobasic phosphates, preferably sodium and / or potassium.

Alternatively, when an infant formula of the invention has a reduced calcium content as described above, the formula of the invention may be prepared with a reduced calcium phosphorus ratio of 1.4 and 2. This reduced calcium-phosphorus ratio may still have some advantages. to compensate for certain water properties used by the end user to prepare the formula, for example by dissolving the powdered formula of the invention.

Also, when one or more prebiotic components C are present in the infant formula of the invention, their content is preferably at least 0.3 g / 100 ml of the total formula, more preferably at least 0.6 g / 100 ml of the total formula. When the formula is given to infants in the usual amounts - for example at least approximately 200 ml per day (depending mainly on the infant's age) as one or more, and preferably 4 or more daily meals - this will correspond to a total daily dose of prebiotic components

13/38 of at least 1.8 g / day (again depending mainly on the age of the infant).

Further, when one or more viscosity enhancing components D are present, the viscosity of the final formula is preferably between 20 and 100 cps, more preferably between 40 and 80 cps; measured on a Brookfield viscometer at 30 rpm.

The infant formula of the invention preferably also has a pH in the range of 4.0 to 7.5.

Still according to a specific embodiment of the invention, the infant formula of the invention may (also) have a low lactose content to prevent and / or reduce problems related to (partial) lactose intolerance and / or reduced lactase activity in the colon. In this embodiment, the infant formula of the invention contains any combination of at least two, preferably at least three, and most preferably the four components from A to D; and has a maximum lactose content of 6.0 g / 100 ml, preferably less than 4.5 g / 100 ml, more preferably less than 3.0 g / 100 ml of the total formula, as determined by the assay of lactose is known per se.

Such a formula may be important for lactose intolerant and / or lactose intolerant infants, for example because they are premature and / or because they have (may have) low lactase activity due to, for example, hereditary factors.

The infant formula of the invention may generally be prepared by mixing / combining various components / ingredients in the amounts mentioned herein. This can generally be performed in a manner known per se using a known mixture and / or equipment for processing it. The formula for infants

It may also be heated / sterilized (with for example UHT treatment) and thereafter preferably taken to a powdered form, with for example evaporation or spray drying; and is aseptically packaged with a suitable container such as a can, box, bag, vial or sachet. A preferred method for preparing the infant formula of the invention from the preferred hydrolysates used in the invention will be discussed in more detail below.

Components A to D and any of the additional components to be used in the invention will also be discussed below. Accordingly, it will be apparent to the skilled artisan that any component or ingredient used in the infant formula of the invention should be a component accepted in the Food Industry, and in particular accepted for use in infant formula. Also, any component (s) or ingredients used should preferably be compatible with the other components / ingredients of the final formula, and more preferably should not (excessively) decrease the value of the desired properties of the final formula, which includes but is not limited to those provided. by any other component / ingredient.

I: Component A:

Component A comprises intact proteins, a protein hydrolyzate, or a combination of the two.

Component A is preferably such that it has a low phosphorus content and is preferably also such that it is easily digestible. Also, preferably, this protein source provides all required amino acids and in sufficient amounts for optimal growth. Suitable sources include specific fragments of mammalian milk protein or protein of low phosphorus origin such as

15/38 soybean. More preferably, for infants who may be intolerant to intact proteins, the proteins should be hydrolyzed.

Preferably, this is a hydrolyzate obtained by hydrolysis of one or more milk proteins, in particular cow's milk, which includes but is not limited to proteins, protein fractions and / or protein preparations derived from whole milk, skimmed milk, casein and / or whey.

In principle, any milk protein hydrolyzate known per se which is suitable for use in food products may be used, more particularly that is suitable for use in the infant formula; and these protein hydrolysates are well known in the art.

Preferably, the protein hydrolyzate used has one or more, and preferably all the characteristics of a) to e) described below. Accordingly, although some prior art methods are not excluded, a protein hydrolyzate having one or more of the desired characteristics of a) to e) should be provided - and these hydrolysates may be used in the infant formula of the invention - the present invention. also provides a range of preferred protein hydrolysates, as well as a method for preparing such hydrolysates, which simultaneously show all the desired characteristics of a) - e). 0 Use of these hydrolysates is particularly preferred.

In particular, a protein hydrolyzate used in the invention should have one or more of the following characteristics:

(a) The hydrolyzate shall contain all amino acids necessary for the good growth of the infant

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b) at least 50% by weight of the peptides present in the hydrolyzate (based on all protein components present in the mixture, including any free amino acids) should have a chain length of 2 to 30 amino acids, preferably 2 to 15 amino acids, and more preferably from 3 to 12 amino acids; the remaining protein components having more amino acids in the protein chain, and / or free amino acids. Preferably, of these remaining protein components, between 0.5 and 8%, more preferably between 0.7 and 7%, and even more preferably between 0.75 and 5%, will have a chain length of greater than 30 amino acids. The presence of small oligopeptides increases protein digestibility, and thus the digestibility of the entire formula. Also, the presence of 15 to 30 amino acid oligopeptides allows the infant's digestive system to inhabit larger and / or intact peptides;

c) a low osmolality due to a low content of free amino acids and salts. This avoids disorders of the gastrointestinal tract / digestive system such as diarrhea, or at least reduces (also) the risk and prevents a low level of stomach emptying. In this regard, it should be noted that, since the infant formula of the invention is particularly intended for infants who have problems with food adaptation / infant formula - for example suffering from one or more of the above problems - it is important that these formulas, if desired, may have a high protein content. This makes controlling the osmolality of the hydrolyzate used to prepare the infant formula of the invention the most important.

Also, as mentioned above, a low free amino acid content is important as free amino acids can give the hydrolyzate - and hence the whole formula - a bitter taste. To mask this bitter taste, it will be necessary to add sweeteners to the formula, in particular sweeteners containing

Many free sugars such as glucose syrup or certain starch hydrolysates (eg corn or peas). These, however, generally have a high osmolality (as starch hydrolysates having a DE value> 35), which in turn will (too) increase the osmolality of the final formula.

Therefore, it is another object of the invention to provide a hydrolyzed infant protein formula which may contain fewer sweeteners and / or sweeteners with a low osmolality (with for example an ED value of less than 30). Preferably, as mentioned above, the osmolality of the hydrolyzate is such that when the hydrolyzate is incorporated into the infant formula in the amounts indicated herein - the osmolality of the final formula is less than 270 mOsmol / 1, preferably less than 240 mOsmol / 1, more preferably less than 230 mOsmol / 1;

d) a good taste, and in particular a non-bitter taste, which is an extremely important feature for (protein hydrolysates to be used in) infant formulas.

The taste, and in particular the bitter taste, of a protein hydrolyzate may for example be determined in a comparative test comprising a taste analysis and the use as a referent of a suitable compound such as caffeine or a standard hydrolyzate.

In particular, in the analysis of the taste test described in Example 2 of US-A-5,837,312 which uses a 5% solution of the compound to be evaluated and which uses as caffeine any hydrolyzate to be used. in the infant formula of the invention it should preferably have a bitter taste of less than 2.7; preferably less than 2.4, more preferably between 1.6 and 2.3 (on a scale of 1 to 6 with 6 being the reference value for caffeine).

As is known in the art, the bitter taste of protein hydrolysates is due (at least in part) to the presence of

18/38 free amino acids that form during hydrolysis, such as free phenylalanine and / or tyrosine). Thus, in the hydrolysates to be used in the invention, the amount of these free amino acids should be as low as possible, ie not more than 10 wt%, and preferably not more than 7 wt% of all protein components (including amino acids). free).

e) a low level of awareness, for example as determined by the tests described by JJ Pahud & K. Swartz in Proc. of the Tropical Conference, Virginia 1985, AOAC, 264-271; and / or Piacentini et al., Allergy 1994, 48, 361-364 or any other suitable test. In particular, the hydrolyzate is such that when the hydrolyzate is incorporated into the infant formula in the amounts indicated herein - in the tests described by Pahud & Swartz or Piacentini the final infant formula of the invention preferably shows an essentially zero degree of sensitization. .

As is known in the art, the allergic properties of a hydrolyzate - and hence the infant formula in which it is incorporated - is in part largely dependent on the type and amount of peptides that form during / remain after hydrolysis. In this regard, it is particularly advantageous to use a protein hydrolyzate prepared according to the method described below.

Although the osmolality imbalances of the protein hydrolyzate may be rectified by the elimination of ions via techniques known in the art, it is very desirable that the hydrolysates fulfill all of the a) to e) characteristics mentioned above. Such a hydrolyzate may be prepared reliably via the method described below. Preferably, the hydrolyzate is present in the infant formula of the invention in an amount between 1 and 3 g / 100 ml, more preferably between 1.3 and 2.0 g / 100 ml, and more preferably between 1.6 and 1.9 g / 100 ml of the total formula.

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II: Method for preparing preferred protein hydrolysates suitable for use in the invention as component A, and the use of said hydrolyzate to prepare the infant formula of the invention:

As mentioned above, this method as well as the hydrolysates obtained with this method form separate aspects of the invention.

As the precursor material for the hydrolysis of the invention any protein, protein mixture or protein preparation having a phosphorus content of less than 0.75 g / 100 g of protein may be used. Particularly preferred is the use of mammalian milk proteins, in particular bovine milk such as whey, in particular acid whey; dephosphorylated casein; β-casein or a suitable mixture thereof.

More preferably, the protein precursor material has a phosphorus content of less than 0.72 g P / 100 g protein, more preferably less than 0.69 g P / 100 g protein. Again, these precursor materials will include whey, in particular acid whey; dephosphorylated casein; or specific casein fractions with a reduced a-casein content and / or an increased β-casein content.

Optionally, the precursor material may be treated to eliminate any native enzymatic activity (residue).

Also, optionally, an optionally homogenized and heated yeast cell suspension and / or pretreated to knead the cell membranes may be added to the precursor material prior to use in the hydrolysis of the invention, for example in an amount of 1-8 ° C. g yeast cell dry mass per 100 g protein. For example, an acid whey mixture containing 4% yeast may be used.

Protein (e.g. added as a 10% baker's yeast suspension).

The precursor material - as such or in a suitable form as a solution or suspension - is then treated with a combination of at least one endo and at least one exoproteinase, wherein said enzyme mixture is added in an amount of 0.1-5% based on the precursor material.

Endo and exoproteinases may be used sequentially for example in two or more separate hydrolysis phases or together, as for example a suitable mixture in a single hydrolysis phase, optionally in combination with one or more additional hydrolysis phases using any one. of the remaining enzymes. Preferably, the hydrolysis comprises a single hydrolysis phase that uses a combination of all enzymes to be used.

Preferably, as endo and exoproteinases, a suitable mixture of serine proteases is used, which includes but is not limited to a suitable enzyme mixture containing (at least) the enzymes Alcalase® and Trypsine® (both available from Novo Nordisk). Another enzyme suitable for use in said enzyme mixture is Flavourzyme® (also available from Novo Nordisk). Particularly preferred is the use of a mixture of Alcalase, Flavourzyme and Trypsine in relative amounts of approximately 5-10 parts Alcalase to 3-5 parts Flavourzyme to 1 part Trypsine (such as a 7.5: 4: 1 mixture ), said mixture is most preferably used in an amount corresponding to at least 0.1% - for example approximately 0.2% - of Trypsine per 100 g of precursor protein.

Using this preferred mixture as described below, a hydrolyzate can be obtained which, despite the presence of relatively high molecular weight peptides / proteins, is not

21/38 sensitizing. Application of the conditions as described above allows the preparation of a formula for infants with a non-bitter taste and low osmolality (as for example described above).

Using a mixture of endo and exoproteinases - and in particular the preferred mixture of Alcalase, Flavourzyme and Trypsine - the precursor material is hydrolyzed at a temperature of approximately 45-60 ° C, preferably between 50 and 58 ° C for less than 4 hours. preferably from 1.5 to 3 hours. During hydrolysis, the pH is kept in the range of 6.4 to 8, and preferably kept essentially constant, in the range of for example 6.8 to 7.8.

Generally, the degree of hydrolysis is not critical. In other words, regardless of the final degree of hydrolysis, hydrolysis of the foregoing precursors using the enzyme combination (s) under the conditions mentioned and over the preceding period usually results in a hydrolyzate with the above conditions. desired properties to be used in the infant formula of the invention. Optionally, if necessary, the skilled artisan will be able - based on this description - to adapt the conditions of the above hydrolysis so that a hydrolyzate with the desired properties is obtained.

In addition, it should be noted that usually - in addition to the use of the foregoing precursors and preferred enzyme combination (s) - hydrolysis conditions may not be too fundamental depending on the manner in which the hydrolyzate is (additionally) ) processed prior to use in the infant formula of the invention. However, it has been found that the desired characteristics of a) to e) obtained in the final infant formula are better and more reliable when the above conditions are used in combination with the cooling phase which will be described below.

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The hydrolysis reaction may be terminated in any suitable manner. However, to ensure the combination of the desired characteristics of a) to e) in the final hydrolyzate, it is more preferred that the hydrolysis reaction is not terminated / cooled by heating (as usual), but by cooling the hydrolysis mixture, for example. without specifically eliminating or denaturalizing the enzyme activities present in the mixture.

In particular, according to the invention, the total reaction mixture is cooled for one hour from the hydrolysis temperature - for example from approximately 45-60 ° C, preferably from 50 to 58 ° C - to a temperature below 20 ° C. C, preferably below 10 ° C. Said cooling is preferably initiated between 1.5 and 3 hours after the start of the hydrolysis reaction.

Thus, a suitable temperature protocol for hydrolysis may be as follows:

- hydrolysis (45-60 ° C, preferably 50-58 ° C): 1.5 to 4 hours

- cooling (to <20 ° C, preferably <10 ° C): 10 min. at 1 hour

- total time: 1.7 to 4.5 hours with the preferred temperature protocol for hydrolysis being as follows:

- hydrolysis (45-60 ° C, preferably 50-58 ° C): 2.0 to 3.0 hours

- cooling (to <20 ° C, preferably <10 ° C): 10 minutes to 1 hour

- total time: 2.2 to 3.5 hours

The cooled hydrolyzate thus obtained may then be used as such in the preparation of the infant formula of the invention, or after suitable treatment, such as by eliminating one or more undesirable compounds or components,

Including but not limited to salts, amino acids, enzymes used in hydrolysis.

This can for example be obtained by desalination (e.g. by dialysis); by precipitation, that is, by acidification with a food acid such as hydrochloric acid to pH <5.0, followed by removal of the precipitate (eg by centrifugation) and readjusting the pH to a suitable value, eg between 6 and 7 ; or by any other suitable technique.

Also, instead of using the entire hydrolyzate, only a fraction thereof can be used, obtained for example by deleting (part) of proteins with a chain length of greater than 30 amino acids.

The hydrolyzate thus obtained is then incorporated into the infant formula of the invention, for example by the following non-limiting treatment steps, and which after the hydrolysis reaction has ended are preferably carried out within 24 hours, more preferably within 8 hours:

a) All other ingredients of the final formula - for example lipid component (s) B; the prebiotic C component (s) (if any); optionally a portion of the viscosity improver (s) D (if any); and optional additional components such as vitamins, minerals and trace elements - are added to the hydrolyzate.

The mixture is then homogenized and heated to the desired microbiological quality of the product without (excessively) diminishing the desired characteristics of the formula and / or the desired biological activity of the components incorporated therein. For example, a UHT treatment at 140 ° C may be used for about 2.4 seconds. Obtained The product is then taken to a powder form - for example by evaporation and drying (by spraying). It is preferred that some ingredients such as

24/38 if there are viscosity enhancing agents D

<td>added</td><td>to dust</td><td>after</td><td>The</td><td>pulverization</td><td>in dry.</td><td>Like this,</td><td>The</td>

<td>composition</td><td>Final</td><td>can</td><td>to be</td><td>packed</td><td colspan="2">aseptically</td><td>on one</td>

<td>container</td><td>appropriate</td><td>how</td><td>per</td><td>example one</td><td>can one</td><td>Cashier,</td><td>an</td>

purse, bottle or sachet.

(b) Alternatively, after the addition of water-soluble components to be incorporated into the final formula - such as vitamins, minerals and trace elements - the hydrolyzate may be heated and partially dried / evaporated to the component (s) are added. s) B lipids and any remaining components. The mixture may then be homogenized and made into a powder form. After optional addition of further powdered ingredients as viscosity enhancing agents, the final composition is packaged, i.e. as described above.

An infant formula prepared in this way - that is via the above mentioned method of the preferred hydrolyzate - has optimal characteristics with regard to taste, low osmolality, low free amino acids, low sensitization characteristics, and protein content. / amino acids which is particularly suitable for the infant formula.

Also, in the method described above, the hydrolyzate is only subjected to minimal heat treatment which ensures that the biological activity of the components present in the formula is maintained, or at least not significantly decreased.

In addition, the above method reliably gives a hydrolyzate and / or infant formula with the desired characteristics, which is produced safely and cost-effectively (limited use of high cost enzymes such as trypsin and low production costs).

25/38

Thus, according to other aspects, the invention relates to a hydrolyzate obtained via the method described in this section II; an infant formula comprising such a protein hydrolyzate; and the use of such a protein hydrolyzate for the preparation of an infant formula, in particular for use as component A in the preparation of an infant formula of the invention.

As stated above, the preferred hydrolyzate is preferably present in the infant formula of the invention in an amount between 1 and 3 g / 100 ml, more preferably between 1.3 and 2.0 g / 100 ml, and most preferably between 1.6 and 1.9 g / 100 ml. ml of the total formula.

However, in addition to its use in the invention, it is to be noted that the method described in this section II - as well as the hydrolysates obtained via said method - may serve other applications, particularly in the field of food products, more particularly in the field. feeding infants and / or dietary foods, for example for the prevention or treatment of food allergies, and the production of foods that are destined to pass quickly to the stomach and which are easily digested. These products can be very useful for patients with poor stomach function, poor liver function, poor pancreatic function and / or poor bowel function.

<td>As</td><td>O</td><td>already mentioned before,</td><td>the method</td><td>of this</td><td>section II</td>

<td>as</td><td>the</td><td>hydrolyzed him</td><td>obtained,</td><td>form</td><td>aspects</td>

<td>independent</td><td>and</td><td>separate from this</td><td>invention.</td><td></td><td></td>

<td colspan="2">III: Component</td><td>B:</td><td></td><td></td><td></td>

<td>At least</td><td>one</td><td>lipid component</td><td>B - this</td><td>it is a</td><td>or more</td>

fat and / or fatty acid component (s) present in the infant formula of the invention - can be chosen from all fatty acid triglycerides known per se

26/38 for use in food products, and are preferably chosen from all fatty acid and / or phospholipid triglycerides (and / or mixtures thereof) known per se for use in infant foods, and in particular those known per se. to be used in infant formulas.

As is well known in the art, these fatty acid triglycerides generally comprise a glyceride molecule to which is attached via spherical bonds three fatty acid residues, which may be the same or different, and which are generally chosen from saturated fatty acids. and unsaturated containing 6 to 26 carbon atoms, including but not limited to linoleic acid, α-linolenic acid, oleic acid, palmitic acid (Ci₆) and / or stearic acid (Ci₈) .

These fatty acid triglycerides may be different from existing fatty acid residues and / or the respective position (s) of the fatty acid residues (for example at position 1, 2 and / or 3). Normally, as is known in the art, a lipid component for use in the infant formula will comprise a suitable mixture of two or more, and usually several different fatty acid triglycerides, depending on for example the desired properties for the component.

<td>lipid fat was</td><td>fat source, obtained.</td><td>like this</td><td>like the way</td><td>where the</td>

<td>At least</td><td>a component B</td><td>lipid</td><td>- this is a</td><td>or more</td>

<td colspan="2">acid triglycerides</td><td>fat</td><td>present in it</td><td>are</td>

preferably chosen so that the lipid component B is easily digestible by the infant, thereby (also) making the final formula easily digestible. This improves the stool structure and increases the biological availability of the bivalent cations as well as calcium and / or fats / fatty acids present in the formula.

27/38

In this regard, the amount and position of the palmitic acid residues present in the triglycerides which are present / represent at least one lipid B component are of particular significance. Preferably, the palmitic acid residues should represent more than 10%, preferably 16

24% of all fatty acid residues in triglycerides used as or in component B.

Also, the amount of triglycerides at which the 04: 0, 06: 0 and / or 08: 0 fatty acids are in the Snl or Sn3 position and in particular where the palmitate residues are in the Snl or Sn3 position should be the lowest. possible. The amount of palmitic acid at the Sn1 or Sn3 position should preferably represent less than 16%, more preferably less than 13%, and most preferably less than 10.7% of all triglycerides present. This corresponds to amounts of triglycerides with the palmitate residue at the Sn1 or Sn3 position less than 0.55 g / 100 ml, preferably less than 0.50 g / 100 ml, more preferably less than 0.38 g / 100 ml of the infant formula of the invention.

In other words - but somewhat less restrictive - when palmitic acid residues represent more than 10%, preferably 16 - 24% of all fatty acid residues present in triglycerides used in or as component B of the invention, of these acid residues. palmitic, as much as possible, and preferably at least 30%, more preferably at least 40%, should be in the 2 position or the β position of the triglyceride.

Mixtures of triglyceride fatty acids containing more than 10%, preferably 16 - 24% palmitate residues, and containing one or two requirements as regards the position of palmitate residues indicated in the preceding two paragraphs,

28/38 will also be referred to below as the preferred lipid component B. These include specific phospholipids as well as so-called structured fats, such as animal or vegetable fats, in which triglycerides have been subjected to enzymatic re-esterification or otherwise such that most of the palmitic acid residues are present. present in heading 2.

These preferred components B are commercially available - for example by Loders Croklaan under the name Betapol® (beta palmitate) - and / or may be prepared in a manner known per se, for example as described in European Patent Applications EP 0 698 078 and / or EP 0 758 846.

However, for better absorption of these fat mixtures into the infant's intestines, the use of a preferred lipid component B as described above may allow the amount of component B that is added to the infant formula of the invention to be reduced thereto. It also provides sufficient energy / nutritional value to the infant.

In particular, with the use of a preferred lipid component B, the overall fat content of the infant formula of the invention may be reduced from 48 wt% or more for conventional infant formulas to less than 46 wt% in an infant formula. of the invention.

Also, the use of a preferred lipid component B may allow the overall calcium content of the infant formula of the invention to be decreased once again as indicated above. This reduction in calcium content can at the same time allow the overall phosphorus content of the infant formula of the invention to be decreased again as indicated above.

Alternatively, the use of a preferred lipid component B may allow the calcium-phosphorus ratio of the formula to

Infants of the invention is decreased, for example from approximately 2 for conventional formulas to a range of approximately 2.0 to 1.4.

Finally, while it has been described that component B consists essentially only of fatty acid triglycerides, it should be understood that the presence of lower amounts of mono- or diglycerides (up to a total of up to one third of all fatty acids of all fatty acid mono-, di- and triglycerides present in or as component B, the remainder being fatty acid triglycerides up to a total of 100% of component B). These mono- or diglycerides usually contain the fatty acid residues as above.

IV: Component C:

One or more prebiotic C components may be chosen from all prebiotic components known per se to be used in food products, and are preferably chosen from among the prebiotic components known per se to be used in infant foods, and in particular among those which are per se are known to be used in infant formulas.

Other preferred but non-limiting examples of these include prebiotic (non-digestible) oligosaccharides. These will usually be hydrolyzed or unhydrolyzed carbohydrates comprising sugar residues interconnected via beta 1-1, 1-2, 1-3, 1-4, 1-6 and / or alpha 1-6 bonds. More preferably, these oligosaccharides will contain from 2-20 sugar residues in the saccharide structure.

For example, these prebiotic oligosaccharides include, but are not limited to, lacto-n-tetraose, lacto-N-fucopentaose, lactose, lacto-sucrose, raffinose, galacto-oligosaccharides, soybean polysaccharide-derived oligosaccharides, mannose-based oligosaccharides, arabinooligosaccharides, xyloligosaccharides, isomaltooligosaccharides, glucans, sialyl oligosaccharides, fucooligosaccharides, and / or any suitable combination thereof.

Lactose may also be used as the prebiotic component. However, the use of lactose in the infant formula of the invention should be avoided when a low lactose formula is desired, for example a formula for infants who are, or may be, lactose intolerant (based for example on age or hereditary factors).

Particularly preferred is the use of one or more transgalactooligosaccharides (TOS), one or more fructooligosaccharides (FOS) and / or mixtures thereof, in particular the use of mixtures of one or more transgalactooligosaccharides and one or more fructooligosaccharides in which the TOS-FOS ratio is from 5: 1 to 15: 1, and more particularly approximately 9: 1, the mixtures of which usually provide a prebiotic effect comparable to that of breast milk.

The amount of one or more prebiotic C components usually used will depend on the specific prebiotic component (s) used, as well as the daily amount of the formula to be administered to the infant, for example as one or more, and preferably four or more daily doses. Generally, the amount of prebiotic component (s) will be such that when the formula is administered to the infant in the recommended / prescribed amount it is sufficient to provide a daily dose of prebiotic component (s) of at least 1.8 g. / day for infants under months of age;

preferably at least 3.6 g / day for infants over the age of months

Usually, to an infant will be 100 ml of daily administered a total of between 500 and formula of the invention. Thus, the previous quantities

31/38 correspond to a quantity of component (s) C of at least

0.3 g / 100 ml of the total formula, more preferably at least 0.6 g / 100 ml of the total formula.

The use of the foregoing prebiotic components for the infant formula of the invention may (help) restore the natural gastrointestinal flora and / or result in a reduction in aggressive fecal components such as components that may irritate or injure epithelial cells. (such as those lining the gastrointestinal tract and / or the skin). The latter is extremely important for infants suffering from diaper dermatitis.

V: Component D:

One or more of the viscosity enhancing components D may be chosen from among all the viscosity enhancing components known per se for use in food products, they are preferably chosen from the viscosity enhancing components known for use in infant foods, especially those known to be used. per se for use in infant formulas; see for example European Patent Application EP 0 846 422.

Some preferred but not limiting examples include carbohydrate based viscosity enhancing components, and in particular those based on starches and / or starch derivatives such as pregelatinized starches, such as pregelatinized potato starch, which is preferred.

The amount of the viscosity enhancer component (s) commonly used will depend upon the viscosity enhancer component (s) used and the other components present in the formula, and will usually be of the same nature as the viscosity. of the final formula is between 20 and

100 cps, preferably between 40 and 80 cps; according to 32/38 measured on a Brookfield viscometer the total amount of the viscosity component (s) will be between 0.5 and preferably between 1 and 3 g / 100 potato pregelatinized can be between 1.4 and 2.1 g / 100 ml.

at 30 rpm. Normally, the enhancer (s) of the 5 g / 100 ml formula, ml. For example, starch used in an amount of

Use of the viscosity enhancer component (s) D may provide several advantages, including but not limited to preventing or reducing regurgitation and / or excessive aerophagia (belching).

VI: Additional Components:

In addition to the above mentioned components A through D, the composition of the invention may additionally contain any component or ingredient for the infant formula known per se, in customary amounts (as for example as described by national or international directives) and depending on the (properties) of the desired final formula. These additional components may include, but are not limited to one or any of the following combinations:

one or more of the following compounds: taurine, choline, carnitine, inositol, biotin;

one or more polyunsaturated long chain fatty acids (PUFAs), including but not limited to linoleic acid, α-linoleic acid, oleic acid, arachidonic acid and docosahexaenoic acid.

- one or more nucleotides and / or nucleotide analogs;

one or more (added) amino acids, including but not limited to tryptophan and / or methionine;

33/38

- one or more (aggregated) vitamins, including but not limited to vitamin K, vitamin B, vitamin B2, vitamin A, vitamin E, vitamin D, vitamin C, niacin, pantothenic acid. In addition to the usual amounts of these or other vitamins, the infant formula of the invention may contain additional amounts of - for example - folic acid, vitamin B12 and / or vitamin B6;

- one or more minerals and / or trace elements;

one or more preservatives (not preferred);

- one or more flavors and / or colors.

Examples:

Example 1: Formula for infants over 4 months old.

An infant formula for infants over 4 months of age is prepared essentially as described above in point II. The formula has the following composition, an energy / nutritional value of 72 kcal / ml and an osmolality of 260 mOsmol / 1. The calcium content is 86 mg / 100 kcal and the phosphorus content is 54 mg / 100 kcal.

Component

Protein equivalents

G whey hydrolyzate per 100 g powder 12.4 g per 100 ml * milk protein ** Carbohydrates

54.8

8.6

Saccharides (lactose)

4.0 (2.7)

Maltodextrine polysaccharides

Starch polysaccharides

Potato 34/38

Organic Acids Fats

Saturated

Beta Palmitate (Croklaan Loders)

Monounsaturated

Polyunsaturated Nondigestible Fiber / Oligosaccharides

TOS

FOS

Additional Components:

Minerals, trace elements and vi by EEC Directive 321.

Notes: *: obtained by dissolving 15 **: hydrolyzate A of point II proteins.

0.1

21.1 3.3 (= 41% by weight)

1.4

0.23

1.4

0.54

5.0 0.78 . 7

0.08 amines in recommended quantities g of powder in 100 ml of water, preferred as described above in

Example 2: Formula for infants up to 6 months old.

A formula for infants for infants up to 6 months old is essentially prepared as described above in point II. The formula has the following composition, a value of

<td colspan="3">energy / nutritional ratio of 70 kcal / ml and an osmolality</td><td rowspan="2">from 2 40 phosphor</td>

<td>mOsmol / 1. The calcium content is is 27 mg / 100kcal.</td><td>53 mg / 100 kcal</td><td>and the content of</td>

<td>Component</td><td>g per 100 g of dust</td><td>g per 100</td><td>ml *</td>

<td>Protein equivalents</td><td> 11.6</td><td> 1. 7</td><td></td>

<td>Whey Hydrolyzate</td><td></td><td> 1. 7</td><td></td>

<td>protein milk **</td><td></td><td></td><td></td>

<td>Carbohydrates</td><td> 55.4</td><td> 8.3</td><td></td>

<td>Saccharides (lactose)</td><td></td><td> 4.0 (2.</td><td> 9)</td>

<td>Polysaccharides</td><td></td><td> 1.5</td><td></td>

<td>from maltodextrine</td><td></td><td></td><td></td>

<td>Starch polysaccharides</td><td></td><td> 1.8</td><td></td>

potato

35/38

Fats

21.1

3.3 (= 42% by weight)

Saturated 1.4

Beta Palmitate 0.23

Monounsaturated 1.4

Polyunsaturated 0.54

Nondigestible fiber / oligosaccharides 5.0 0.78

TOS 0.7

FOS 0.08

Additional Components:

Minerals, trace elements and vitamins in amounts recommended by EEC Directive 321.

Notes: *: Obtained by dissolving 15.0 g of powder in 100 ml of water.

**: Preferred protein hydrolyzate A as described above under II.

Example 3: Formula for infants up to 4 months old.

A formula for infants for infants up to 4 months old is essentially prepared as described above in point II. The formula has the following composition, a value of

<td>energy / nutritional</td><td>kcal / ml and one</td><td>osmolality</td><td>from 23 0</td>

<td>mOsmol / 1. The calcium content is</td><td>45 mg / 100 kcal</td><td>and the content of</td><td>phosphor</td>

<td>is 25 mg / 100kcal.</td><td></td><td></td><td></td>

<td>Component</td><td>g per 100 g of dust</td><td>g per 100</td><td>ml *</td>

<td>Protein equivalents</td><td> 10.5</td><td> 1.54</td><td></td>

<td>Whey Hydrolyzate</td><td></td><td> 1.46</td><td></td>

<td>protein milk **</td><td></td><td></td><td></td>

<td>L-Tryptophan</td><td></td><td> 0.01</td><td></td>

<td>L-methionine</td><td></td><td> 0.01</td><td></td>

<td>Baker's yeast</td><td></td><td> 0.06</td><td></td>

<td>hydrolyzed * *</td><td></td><td></td><td></td>

<td>Carbohydrates</td><td> 55.4</td><td> 8.3</td><td></td>

<td>Saccharides (lactose)</td><td></td><td> 3.5</td><td></td>

<td>Polysaccharides</td><td></td><td> 3.3</td><td></td>

<td>from maltodextrine</td><td></td><td></td><td></td>

<td>Starch polysaccharides</td><td></td><td> 1.4</td><td></td>

Potato 36/38

Fats

Saturated

Beta palmitate

Monounsaturated

Polyunsaturated Nondigestible Fiber / Oligosaccharides

TOS

FOS

Lacto-N-Tetraose Additional Components:

Folic acid (pg)

Vitamin K (pg)

Minerals, trace elements and EEC Directive 321

19.2 3.0 (= 40% by weight)

1.2

0.23

1.3

0.5

3.2 0.5

0.37

0.03

0.10 vitamins in recommended amounts

Notes: *: Obtained by dissolving 15.0 g of powder in 100 ml of water.

**: Preferred protein hydrolyzate A as described above under II.

Example 4: Comparative Studies

The effects of the formula of example 2 were tested in a study involving 51 full-term newborns and were compared with the effects of a commercial standard product that did not contain beta palmitate and did not have an adapted calcium-phosphorus ratio. After prolonged feeding with the formulas of the invention and comparing them with the Standard formula, significantly fewer problems with constipation (p &lt;0.05), abdominal discomfort (p &lt;0.005) and fewer gastrointestinal problems (p &lt; 0.01), as well as fewer problems related to gastrointestinal discomfort (spasms and crying).

Lisbon, July 19, 2010

37/38

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the Holder is intended solely to assist the reader and is not part of the European patent document. Even though the utmost care has been taken in its preparation, errors or omissions cannot be excluded and EPO assumes no responsibility in this regard.

Patent Application Documents referenced in the description • US 5837315 A · EP 0758846 A • EP 0698078 A · EP 0846422 A

Literature Cited in Non-Patent Application Description • Genzel-Boroviczény et al. Eur. J. Pediatr., 1997, Vol. 156, 142-147 • Obermeier et al. Isotopes Environ. Health Stud., 1999, Vol. 35, 119-125 • Piacentini et al. Allergy 1994, vol. 48, 361-364 • Piacentini. Allergy 1994, vol. 48, 361-364