Infant Formula

Abstract

Infant Formula An infant formula for use in reducing the incidence of infections in infants or preventing infections in infants, wherein the infant formula is an extensively hydrolyzed infant formula (eHF) or an amino acid-based infant formula (AAF), the infant formula comprising protein, carbohydrates, and lipids, the eHF comprising no more than about 2.4 g of protein per 100 kcal or the AAF comprising no more than about 2.9 g of protein per 100 kcal, the infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and no more than about 30% by weight of the lipids are medium-chain triglycerides (MCT).

Description

[Technical field]

[0001] The present invention relates to an extensively hydrolyzed infant formula (eHF) and an amino acid based infant formula (AAF) containing reduced amounts of protein and containing human milk oligosaccharides (HMO) for use in reducing the incidence of or preventing infectious diseases. The formulas may be used for infants with cow's milk protein allergies. [Background technology]

[0002] Cow's milk protein (CMP) is the leading cause of food allergy in infants, affecting 2-3% of children worldwide. Most children with CMP allergy (CMPA) have two or more symptoms: 50-70% have skin symptoms; 50-60% have gastrointestinal symptoms; and 20-30% have respiratory tract symptoms. In 10% of children, severe, life-threatening symptoms may develop. (Nutten, 2018. EMJ Allergy Immunol, 3(1), pp.50-59).

[0003] Human breast milk and breastfeeding are considered the optimal form of nutrition for healthy infants in the first few months of life. Breast milk remains the gold standard for feeding infants with CMPA. The European Society for Pediatric Gastroenterology and Hepatology (ESPGHAN) recommends that in breastfed infants, the best treatment for CMPA is the complete elimination of cow's milk from the maternal diet (Koletzko, S., et al., 2012. Journal of pediatric gastroenterology and nutrition, 55(2), pp.221-229).

[0004] If breastfeeding is not possible, specialized infant formulas are recommended. ESPGHAN recommends that non-breastfed infants with CMPA use extensively hydrolyzed protein-based formulas, which have been proven effective in infants with CMPA. In infants with very severe or life-threatening symptoms, amino acid-based infant formulas (AAF) may be the first choice (Koletzko, S., et al., 2012. Journal of pediatric gastroenterology and nutrition, 55(2), pp. 221-229).

[0005] Extensively hydrolyzed infant formulas (eHFs) and AAF may have lower nitrogen absorption rates than complete protein formulas or human breast milk (Rigo, J., et al., 1995. European journal of clinical nutrition, 49, pp. S26-38). Thus, to accommodate infant needs, eHFs typically contain 2.6-2.8g protein per 100 kcal, and AAFs typically contain 2.8-3.1g protein per 100 kcal (Borschel, M., et al., 2018. Nutrients, 10(3), p. 289).

[0006] However, consumption of high-protein infant formulas has been associated with increased body weight and body mass index at age 2 years, as well as increased circulating concentrations of plasma essential amino acids, insulin-like growth factor-1, and C-peptide, which may induce weight gain and lipogenic activity. Lower protein content may reduce the risk of subsequent obesity (Totzauer, M., et al., 2018. Obesity, 26(7), pp. 1203-1210).

[0007] Furthermore, infants with CMP allergy (CMPA) may be at higher risk of infection. For example, infants with CMPA are known to have a higher prevalence of respiratory tract infections (Woicka-Kolejwa, K., et al., 2016. Advances in Dermatology and Allergology, 33(2), p. 109). CMPA is also associated with recurrent otitis media in childhood (Juntti, H., Acta Oto-Laryngologica, 119(8), pp. 867-873).

[0008] [Summary of the invention] The inventors have surprisingly found that eHF and AAF, which contain the human milk oligosaccharides (HMOs) 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), contain reduced amounts of protein and have a protein content closer to that of human breast milk, support proper growth and development in infants with CMPA, are safe and tolerable, and reduce the incidence of infections.

[0009] Thus, in one aspect, the invention provides an infant formula for use in reducing the incidence of infections in infants or preventing infections in infants, the infant formula being an extensively hydrolyzed infant formula (eHF) or an amino acid based infant formula (AAF), the infant formula comprising protein, carbohydrate, and lipid, the eHF comprising about 2.4 g or less of protein per 100 kcal, or the AAF comprising about 2.9 g or less of protein per 100 kcal, the infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglycerides (MCT).

[0010] In another related aspect, the invention provides a method of reducing the incidence of infection in an infant or preventing an infection in an infant, the method comprising administering to an infant an infant formula, the infant formula being eHF or AAF, the infant formula comprising protein, carbohydrate, and lipid, the eHF comprising about 2.4 g or less of protein per 100 kcal or the AAF comprising about 2.9 g or less of protein per 100 kcal, the infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglycerides (MCT).

[0011] In another aspect, the present invention provides an infant formula for use in treating an infection in an infant, the infant formula being eHF or AAF, the infant formula comprising protein, carbohydrate, and lipid, the eHF comprising about 2.4 g or less of protein per 100 kcal or the AAF comprising about 2.9 g or less of protein per 100 kcal, the infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglycerides (MCT).

[0012] In one aspect, the present invention provides an AAF for use in reducing the incidence of infections in infants or preventing infections in infants, the AAF comprising protein, carbohydrate and lipid, the AAF comprising about 2.8g or less of protein per 100 kcal, preferably 2.7g or less of protein per 100 kcal, more preferably 2.6g or less of protein per 100 kcal, the AAF further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglyceride (MCT).Suitably, the AAF may comprise 2.5g or less of protein per 100 kcal.

[0013] In another related aspect, the present invention provides a method for reducing the incidence of infections in infants or preventing infections in infants, the method comprising administering an AAF to an infant, the AAF comprising protein, carbohydrate, and lipid, the AAF comprising about 2.8g or less of protein per 100 kcal, preferably 2.7g or less of protein per 100 kcal, more preferably 2.6g or less of protein per 100 kcal, the AAF further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglyceride (MCT).Suitably, the AAF may comprise 2.5g or less of protein per 100 kcal.

[0014] In another related aspect, the invention provides a method of treating an infection in an infant, the method comprising administering to the infant an infant formula, the infant formula being eHF or AAF, the infant formula comprising protein, carbohydrate, and lipid, the infant formula comprising about 2.4 g or less of protein per 100 kcal, the infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglycerides (MCT).

[0015] In another aspect, the present invention provides an AAF for use in treating infectious diseases in infants, the AAF comprising protein, carbohydrate and lipid, the AAF comprising about 2.9g or less of protein per 100 kcal, preferably 2.8g or less of protein per 100 kcal, 2.7g or less of protein per 100 kcal, more preferably 2.6g or less of protein per 100 kcal, the AAF further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglyceride (MCT).Suitably, the AAF may comprise 2.5g or less of protein per 100 kcal.

[0016] In another related aspect, the present invention provides a method for treating an infection in an infant, comprising administering an AAF to the infant, the AAF comprising protein, carbohydrate and lipid, the AAF comprising about 2.9g or less of protein per 100 kcal, preferably about 2.8g or less of protein per 100 kcal, 2.7g or less of protein per 100 kcal, more preferably about 2.6g or less of protein per 100 kcal, the AAF further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and about 30% or less by weight of the lipid is medium chain triglyceride (MCT).Suitably, the AAF may comprise 2.5g or less of protein per 100 kcal.

[0017] Preferably the infant has a cow's milk protein allergy.

[0018] The infection may be an ear infection, a respiratory infection, a gastrointestinal infection, and / or a urinary tract infection. Preferably, the infection is a respiratory infection, such as an upper respiratory tract infection and / or a lower respiratory tract infection. Preferably, the infection is an ear infection, more preferably a middle ear infection.

[0019] The infant formula may include 2'FL and LNnT. The infant formula may include 0.5-3g / L, 0.8-1.5g / L, or about 1g / L of 2'FL, preferably the infant formula includes about 1g / L of 2'FL; and / or the infant formula may include 0.2-1g / L, 0.5-0.8g / L, or about 0.5g / L of LNnT, preferably the infant formula includes about 0.5g / L of LNnT. Most preferably, the infant formula includes about 1g / L of 2'FL and about 0.5g / L of LNnT.

[0020] The infant formula may contain 1.8-2.4 g protein per 100 kcal, 2.1-2.3 g protein per 100 kcal, or 2.15-2.25 g protein per 100 kcal. Preferably, the infant formula contains about 2.2 g protein per 100 kcal.

[0021] About 30% or less, about 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less by weight of the lipids in the infant formula can be medium chain triglycerides (MCT). Preferably, the infant formula does not contain added MCTs.

[0022] Infant formulas may contain 9-14 g carbohydrate per 100 kcal and / or 4.0-6.0 g fat per 100 kcal.

[0023] In one embodiment, the infant formula is eHF. In one embodiment, the infant formula is AAF. Preferably, the infant formula is eHF.

[0024] At least about 95%, at least about 98%, at least about 99%, or about 100% by weight of the peptides in the eHF may have a molecular weight of less than about 3000 Da. Preferably, there are no detectable peptides in the eHF greater than or equal to about 3000 Da in size.

[0025] At least about 90%, at least about 95%, at least about 98%, or at least about 99% by weight of the peptides in the eHF may have a molecular weight of less than about 1500 Da. Preferably, at least about 99% of the peptides in the eHF have a molecular weight of less than about 1500 Da.

[0026] At least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% by weight of the peptides in the eHF may have a molecular weight of less than about 1200 Da. Preferably, at least 98% by weight of the peptides have a molecular weight of less than about 1200 Da.

[0027] At least about 45%, at least about 50%, 45-55%, or 50-54% by weight of the peptides in the eHF can be di- and tripeptides. Preferably, about 51-53%, or more preferably about 52% by weight of the peptides in the eHF are di- and tripeptides.

[0028] At least about 45% by weight, at least about 50% by weight, 45-55% by weight, or 50-54% by weight of the peptides in the eHF can have a molecular weight of 240-600 Da. Preferably, about 51-53% by weight, or more preferably about 52% by weight, of the peptides in the eHF have a molecular weight of 240-600 Da.

[0029] At least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the protein in the eHF can be whey protein. Preferably, the source of protein is whey protein.

[0030] The eHF may contain free amino acids. The free amino acids may be present at a concentration of 50% by weight or less, 40% by weight or less, 30% by weight or less, or 25% by weight or less, based on the total weight of the amino acids. Preferably, the free amino acids are present at a concentration of 20-25% by weight, 21-23% by weight, or about 22% by weight, based on the total weight of the amino acids. [Brief description of the drawings]

[0031] [Figure 1] Non-inferiority: daily weight gain Treatment effect on weight gain at visit 4. The primary analysis shows that weight gain [g / d] of infants fed the test formula was non-inferior to growth on the control formula. [Diagram 2] Odds ratios: Respiratory and ear infections Odds ratios for lower respiratory tract infection (LTRI), upper respiratory tract infection (UTRI), and otitis media / middle ear infection (OM) comparing groups fed test formula (HMO) or control formula. The incidence of OM was significantly lower in the groups fed test formula between V0-V4 and V0-V6. [Diagram 3] Relative risk reduction: respiratory and ear infections Relative risk reduction for lower respiratory tract infections, upper respiratory tract infections, and middle ear infections in infants fed the test formula. The relative risk reduction for middle ear infections was significant in the group fed the test formula. [Figure 4] Odds ratios: medication use (antibiotics, antipyretics) Odds ratios for antibiotic and antipyretic use comparing groups fed test formula (HMO) or control formula. Antipyretic use was significantly lower in the test formula group between V4 and V6. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Various preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

[0033] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0034] As used herein, the terms "comprising," "comprises," and "comprised of" are synonymous with "including" or "includes" or "containing" or "contains" and are inclusive, i.e., open-ended, and do not exclude additional, unrecited components, elements, or steps. The terms "comprising," "comprises," and "comprised of" also include the term "consisting of."

[0035] As used herein, the term "about" means approximately, in the region of, roughly, or in the vicinity of. When the term "about" is used in conjunction with a numerical value or range, the value or range modifies the value or range by expanding the boundaries above and below the numerical value(s) set forth. In general, the terms "about" and "approximately" are used herein to adjust a numerical value(s) above and below the stated value(s) by 10%.

[0036] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Any publications cited herein should not be construed as an admission that they constitute prior art to the claims appended hereto.

[0037] The present disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present disclosure. Numerical ranges are inclusive of the numbers defining the range.

[0038] Infant Formula The term "infant formula" may refer to a food for a specific nutritional purpose for infants during the first year of life, which formula itself meets the nutritional requirements of humans falling within said category (as defined in Commission Regulation (EU) 2016 / 127 of 25 September 2015).

[0039] The infant formula of the present invention is an extensively hydrolyzed infant formula (eHF) or an amino acid based infant formula (AAF). Preferably, the infant formula is an eHF.

[0040] The term "extensively hydrolyzed infant formula" or "eHF" can refer to an infant formula that includes extensively hydrolyzed proteins. An eHF can be a hypoallergenic infant formula that provides complete nutrition for infants who cannot digest intact CMP or who have an intolerance or allergy to CMP.

[0041] The term "amino acid-based infant formula" or "AAF" may refer to an infant formula that contains only free amino acids as a source of protein. The AAF may not contain detectable peptides. The AAF may be a hypoallergenic infant formula that provides complete nutrition for infants with food protein allergies and / or food protein intolerances. For example, the AAF may be a hypoallergenic infant formula that provides complete nutrition for infants who cannot digest intact CMP or who have intolerance or allergies to CMP and who may have very severe or life-threatening symptoms and / or sensitization to multiple foods.

[0042] A "hypoallergenic" composition is one that is unlikely to cause an allergic reaction. Preferably, the infant formula of the present invention is tolerated by more than 90% of infants with CMPA. This is in line with the guidance provided by the American Academy of Pediatrics (Committee on Nutrition, 2000. Pediatrics, 106(2), pp.346-349). Preferably, the infant formula of the present invention may not contain peptides that are recognized by CMP-specific IgE, e.g., IgE in subjects with CMPA.

[0043] Infants can be fed infant formula alone, or infant formula can be used as a supplement to human breast milk.

[0044] The infant formula of the present invention may be in powder or liquid form.

[0045] The liquid may be, for example, a concentrated liquid infant formula or a ready-to-feed infant formula. The infant formula may be in the form of a reconstituted infant formula (i.e., a liquid infant formula reconstituted from a powder form). The concentrated liquid infant formula may preferably be diluted, for example by adding water, to a liquid composition suitable for feeding to an infant.

[0046] In one embodiment, the infant formula is in powder form, which can be reconstituted, for example by adding water, into a liquid composition suitable for feeding to an infant.

[0047] The infant formula, when formulated as directed, may have an energy density of about 60-72 kcal per 100 mL. Preferably, the infant formula, when formulated as directed, may have an energy density of about 60-70 kcal per 100 mL.

[0048] Human Milk Oligosaccharides The infant formula of the present invention contains one or more human milk oligosaccharides (HMOs).

[0049] Many types of HMOs are found in human milk. Human milk contains a great variety of oligosaccharides, with more than 130 such structures identified so far, since each oligosaccharide is based on a combination of glucose, galactose, sialic acid (N-acetylneuraminic acid), fucose, and / or N-acetylglucosamine with numerous and varied linkages between them. Almost all of these oligosaccharides have a lactose residue at the reducing end, and sialic acid and / or fucose (if present) occupy the terminal positions of the non-reducing end. HMOs can be acidic (e.g., charged sialic acid-containing oligosaccharides) or neutral (e.g., fucosylated oligosaccharides).

[0050] The infant formula of the present invention contains 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT).

[0051] The infant formula of the present invention may include 2'FL. In some embodiments, there are no other types of fucosylated oligosaccharides than 2'FL, i.e., the infant formula of the present invention includes only 2'FL as a fucosylated oligosaccharide.

[0052] 2'FL can be produced by biotechnological means using specific fucosyltransferases and / or fucosidases, either using enzyme-based fermentation techniques (recombinant or natural enzymes) or microbial fermentation techniques. In the latter case, microorganisms can be engineered to express any of those natural enzymes and substrates or to produce the respective substrates and enzymes. Alternatively, 2'FL can be produced by chemical synthesis from lactose and free fucose.

[0053] The infant formula of the present invention may include LNnT. In some embodiments, there are no other types of N-acetylated oligosaccharides than LNnT, i.e., the infant formula of the present invention includes only LNnT as an N-acetylated oligosaccharide.

[0054] LNnT can be chemically synthesized by enzymatic transfer of a sugar unit from a donor moiety to an acceptor moiety using a glycosyltransferase, for example as described in U.S. Pat. No. 5,288,637 and WO 96 / 10086. Alternatively, LNnT can be prepared by chemical conversion of a ketohexose (e.g., fructose) that is free or bound to an oligosaccharide (e.g., lactulose) to an N-acetylhexosamine or an N-acetylhexosamine-containing oligosaccharide, as described in Wrodnigg, TM; Stutz, AE (1999) Angew. Chem. Int. Ed. 38:827-828. The N-acetyl-lactosamine thus generated can then be transferred to lactose as the acceptor moiety.

[0055] In some embodiments, the infant formula of the present invention comprises an oligosaccharide mixture comprising 2'FL and / or LNnT. In a preferred embodiment, the infant formula of the present invention comprises an oligosaccharide mixture comprising 2'FL and LNnT. The infant formula of the present invention may comprise only 2'FL as the fucosylated oligosaccharide and only LNnT as the N-acetylated oligosaccharide.

[0056] 2'FL may be present in the infant formula according to the invention in a total amount of 0.5-3g, e.g., 0.8-1.5g, per liter of infant formula (when formulated as directed). In some embodiments, the total amount of 2'-fucosyllactose may be 0.85-1.3g, e.g., 0.9-1.25g or 0.9-1.1g or 1-1.25g or 1-1.2g, per liter of infant formula (when formulated as directed). Preferably, the infant formula (when formulated as directed) contains about 1g / L of 2'-fucosyllactose.

[0057] LNnT may be present in an infant formula according to the invention in a total amount of 0.2-1 g, e.g., 0.5-0.8 g, per L of infant formula (when formulated as directed). In some embodiments, the total amount of LNnT may be 0.5-0.75 g, 0.5-0.7 g, or 0.5-0.6 g, per L of infant formula (when formulated as directed). Preferably, the infant formula (when formulated as directed) contains about 0.5 g / L of LNnT.

[0058] All of these different ranges can be combined together.

[0059] Thus, in one embodiment of the present invention, an infant formula (when formulated as directed) comprises 2'FL and LNnT, (i) the total amount of 2'FL is between 0.8 and 1.5 g per liter of infant formula; and / or (ii) The total amount of LNnT is 0.5–0.8 g per L of infant formula.

[0060] In another embodiment, an infant formula of the invention (when formulated as directed) comprises 2'FL and LNnT, (i) the total amount of 2'FL is between 0.9 and 1.25 g per liter of infant formula; and / or (ii) The total amount of LNnT is 0.5–0.7 g per L of infant formula.

[0061] In another embodiment, an infant formula of the invention (when formulated as directed) comprises 2'FL and LNnT, (i) the total amount of 2'FL is between 1 and 1.2 g per liter of infant formula; and / or (ii) The total amount of LNnT is 0.5-0.6 g per liter of infant formula.

[0062] In a preferred embodiment, an infant formula of the invention (when formulated as directed) contains about 1 g / L 2'FL and about 0.5 g / L LNnT.

[0063] The infant formula of the present invention may contain 0.075-0.5 g / 100 kcal, 0.1-0.3 g / 100 kcal, or 0.12-0.25 g / 100 kcal of 2'FL and about 0.03-0.15 g / 100 kcal, 0.05-0.12 g / 100 kcal, or 0.05-0.1 g / 100 kcal of LNnT. Preferably, the infant formula of the present invention contains about 0.15 g / 100 kcal of 2'FL and about 0.075 g / 100 kcal of LNnT.

[0064] The 2'FL and LNnT contained in the infant formula according to the invention are typically present in a ratio of 2'FL:LNnT of 2.0:0.54 to 2.0:2.26, for example 2.0:0.76 to 2.0:1.8 or 2.0:0.8 to 2.0:1.4. In a particularly advantageous embodiment, this ratio is 2.0:1 or about 2.0:1.

[0065] protein The term "protein" includes peptides and free amino acids. The protein content of an infant formula may be calculated by any method known to those skilled in the art. Preferably, the protein content may be calculated by the nitrogen protein equivalent method, for example as described in Maubois, JL and Lorient, D., 2016. Dairy science & technology, 96(1), pp. 15-25. Preferably, the protein content is calculated as the nitrogen content x 6.25, as defined in European Commission Regulation (EU) 2016 / 127 of 25 September 2015. The nitrogen content may be determined by any method known to those skilled in the art. For example, the nitrogen content may be measured by the Kjeldahl method.

[0066] Protein concentration eHF typically contains 2.6-2.8 g of protein per 100 kcal, and AAF typically contains 2.8-3.1 g of protein per 100 kcal, to address the needs of infants with gastrointestinal pathology, including severe malabsorption, or who require more protein and calories to compensate for a high metabolic rate.

[0067] The inventors have surprisingly shown that eHF or AAF with lower protein content can support proper growth and development in allergic infants. Furthermore, the inventors have surprisingly shown that the formulas are safe and tolerable.

[0068] Thus, an eHF of the present invention contains about 2.4 g or less of protein per 100 kcal, or an AAF of the present invention contains about 2.9 g or less of protein per 100 kcal, preferably about 2.8 g or less of protein per 100 kcal. For example, an infant formula of the present invention may contain about 2.3 g or less of protein per 100 kcal, 2.25 g or less of protein per 100 kcal, or 2.2 g or less of protein per 100 kcal.

[0069] Suitably, the infant formula contains about 1.8g or more of protein per 100 kcal. For example, the infant formula of the present invention may contain about 1.86g or more of protein per 100 kcal, 1.9g or more of protein per 100 kcal, 2.0g or more of protein per 100 kcal, or 2.1g or more of protein per 100 kcal. Preferably, the infant formula contains about 1.86g or more of protein per 100 kcal in line with current EU regulations (EFSA NDA Panel, 2014. EFSA journal, 12(7), 3760).

[0070] An eHF of the present invention may contain 1.8-2.4 g protein per 100 kcal, 1.86-2.4 g protein per 100 kcal, 1.9-2.4 g protein per 100 kcal, 2.0-2.4 g protein per 100 kcal, 2.0-2.3 g protein per 100 kcal, 2.1-2.3 g protein per 100 kcal, or 2.15-2.25 g protein.

[0071] The AAF of the present invention may contain 1.8-2.9 g protein per 100 kcal, 1.9-2.8 g protein per 100 kcal, 2.0-2.7 g protein per 100 kcal, 2.0-2.6 g protein per 100 kcal, 2.0-2.5 g protein per 100 kcal, 2.0-2.4 g protein per 100 kcal, 2.1-2.3 g protein per 100 kcal, or 2.15-2.25 g protein per 100 kcal.

[0072] Preferably, an infant formula of the invention comprises 2.0-2.4 g protein per 100 kcal, such as 2.1-2.3 g protein per 100 kcal, or 2.15-2.25 g protein per 100 kcal.

[0073] Preferably, the infant formula contains about 2.2 g of protein per 100 kcal.

[0074] Protein Source The source of protein can be any source suitable for use in infant formulas. Preferably, the protein is cow's milk protein.

[0075] The highly hydrolyzed / hydrolyzed whey-based formula may be more palatable than the highly hydrolyzed / hydrolyzed casein-based formula, and / or the subject may be sensitized only to casein protein.Therefore, preferably, more than about 50%, more than about 60%, more than about 70%, more than about 80%, more than about 90%, or about 100% of the protein is whey protein.Preferably, the source of protein is whey protein.

[0076] The whey protein may be whey derived from cheese manufacture, in particular sweet whey such as that obtained by coagulation of casein with rennet, acid whey obtained by coagulation of casein with acid or by acidification of a ferment, or even mixed whey obtained by coagulation with acid and with rennet. The raw material may be whey that has been desalted by ion exchange and / or by electrodialysis, known as desalted whey protein (DWP).

[0077] The source of whey protein can be sweet whey from which caseino-glycomacropeptide (CGMP) has been completely or partially removed. This whey is called denatured sweet whey (MSW). Removal of CGMP from sweet whey results in a protein material with threonine and tryptophan content close to that of human breast milk. The process of removing CGMP from sweet whey is described in EP 880902.

[0078] The whey protein may be a mixture of DWP and MSW.

[0079] In some embodiments, the amount of casein in the infant formula is undetectable, e.g., less than 0.2 mg / kg. The amount of casein may be determined by any method known to one of skill in the art.

[0080] Degree of hydrolysis In eHF, proteins are "extensively hydrolyzed," so that eHF can be tolerated by more than 90% of infants with CMPA.

[0081] Protein hydrolysates may have a degree of hydrolysis characterized by NPN / TN%, which refers to non-protein nitrogen divided by total nitrogen x 100. Non-protein nitrogen refers to amino nitrogen that is free to react with reagents such as trinitrobenzenesulfonic acid (TNBS). NPN / TN% may be determined by any method known to those skilled in the art. For example, NPN / TN% may be measured as described in Adler-Nissen (Adler-Nissen, J. (1979) J. Agric. Food Chem. 27:1256-1262). Suitably, proteins may have an NPN / TN% of greater than 90%, greater than 95%, or greater than 98%.

[0082] The degree of hydrolysis can also be determined by the degree of hydrolysis. "Degree of hydrolysis" (DH) is defined as the percentage of peptide bonds broken in a protein hydrolysate and can be determined by any method known to those skilled in the art. Suitably, the degree of hydrolysis is determined by pH stat, trinitrobenzenesulfonic acid (TNBS), o-phthaldialdehyde (OPA), trichloroacetic acid soluble nitrogen (SN-TCA), or formol titration. (Rutherfurd, SM, 2010. Journal of AOAC International, 93(5), pp.1515-1522). The degree of hydrolysis (DH) of a protein can be greater than 90, greater than 95, or greater than 98.

[0083] The degree of hydrolysis can also be determined by peptide molecular weight distribution. The peptide molecular weight distribution can be determined by high performance size exclusion chromatography, optionally with a UV detector (HPSEC / UV) (Johns, PW, et al., 2011. Food chemistry, 125(3), pp.1041-1050). For example, the peptide molecular weight distribution can be an estimate based on HPSEC peak area determined at 205 nm, 214 nm, or 220 nm. Suitably, when the peptide molecular weight distribution is determined by HPSEC / UV, the "weight percent of peptides" having a particular molecular weight can be estimated by the "fraction of the peak area as a percentage of the total peak area" having a molecular weight determined at 205 nm, 214 nm, or 220 nm. Suitably, the degree of hydrolysis can be determined by the method described in WO2016 / 156077. Alternatively, peptide molecular weight distribution can be determined by any method known to those skilled in the art, for example, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Chauveau, A., et al., 2016. Pediatric Allergy and Immunology, 27(5), pp. 541-543).

[0084] Theoretically, a peptide needs to be larger than about 1500 Da (about 15 amino acids) in size to bind to cell membrane-bound IgE, and larger than about 3000 Da (about 30 amino acids) in size to crosslink IgE molecules and induce an immune response (Nutten, 2018. EMJ Allergy Immunol, 3(1), pp.50-59).

[0085] Thus, suitably, at least about 95%, at least about 98%, at least about 99%, or about 100% by weight of the peptides in the eHF have a molecular weight of less than about 3000 Da. There may be no detectable peptides in the eHF greater than or equal to about 3000 Da in size.

[0086] Thus, suitably, at least about 95%, at least about 98%, at least about 99%, or about 100% by weight of the peptides in the eHF have a molecular weight of less than about 1500 Da. Preferably, at least 99% by weight of the peptides have a molecular weight of less than about 1500 Da. There can be no detectable peptides in the eHF of a size greater than or equal to about 1500 Da.

[0087] Preferably, at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% by weight of the peptides in the eHF have a molecular weight of less than about 1200 Da. More preferably, at least 95% or 98% by weight of the peptides in the eHF have a molecular weight of less than about 1200 Da.

[0088] Suitably, at least about 80%, at least about 85%, at least about 90%, or at least about 95% by weight of the peptides in the eHF have a molecular weight of less than about 1000 Da. Preferably, at least about 95% by weight of the peptides in the eHF have a molecular weight of less than about 1000 Da.

[0089] Preferably, the eHF of the present invention has no detectable peptides greater than about 3000 Da in size, and at least about 95% by weight of the peptides have a molecular weight less than about 1200 Da.

[0090] A high proportion of di- and tripeptides may improve nitrogen (protein) absorption in patients with intestinal dysfunction. PEPT1 is a dedicated transport pathway that facilitates the absorption of small peptides (e.g., di- and tripeptides). In the first few weeks of life, intestinal PEPT1 is important for nutrient intake and later for food transition after weaning.

[0091] Thus, at least about 30%, at least about 40%, or at least about 50% by weight of the peptides in the eHF can be di- and tripeptides. Preferably, at least about 45%, at least about 50%, 45-55%, or 50-54% by weight of the peptides in the eHF are di- and tripeptides. More preferably, about 51-53%, or more preferably about 52% by weight of the peptides in the eHF are di- and tripeptides.

[0092] Suitably, at least about 30%, at least about 40%, or at least about 50% by weight of the peptides in the eHF have a molecular weight of 240-600 Da. Preferably, at least about 45%, at least about 50%, 45-55%, or 50-54% by weight of the peptides in the eHF have a molecular weight of 240-600 Da. More preferably, about 51-53%, or most preferably about 52% by weight of the peptides in the eHF have a molecular weight of 240-600 Da.

[0093] The peptides in the eHF may have a median molecular weight of between 300 Da and 370 Da, preferably between 320 Da and 360 Da.

[0094] The major recognized allergens in cow's milk are α-lactalbumin (aLA), β-lactoglobulin (bLG), and bovine serum albumin (BSA).

[0095] Thus, preferably, eHF can have an undetectable aLA content, for example, about 0.010 mg / kg or less aLA, eHF can have an undetectable bLG content, for example, about 0.010 mg / kg or less bLG, and / or eHF can have an undetectable BSA content, for example, about 0.010 mg / kg or less BSA.Preferably, eHF of the present invention does not contain detectable amounts of aLA, bLG, and BSA. The contents of aLA, bLG, and BSA can be determined by any method known to those skilled in the art, for example, ELISA.

[0096] In a preferred embodiment, the eHF of the present invention has no detectable peptides greater than about 3000 Da in size; at least about 95% by weight of the peptides have a molecular weight of less than about 1200 Da; optionally, at least about 45%, at least about 50%, or 45-55% by weight of the peptides have a molecular weight of 240-600 Da and / or are dipeptides or tripeptides; the eHF of the present invention contains about 1 g / L 2'-fucosyllactose and about 0.5 g / L lacto-N-neotetraose, and / or about 0.15 g / 100 kcal 2'-fucosyllactose and about 0.075 g / 100 kcal lacto-N-neotetraose; and the eHF does not contain added MCT.

[0097] Hydrolysis method Proteins for use in the infant formulas of the present invention may be hydrolyzed by any suitable method known in the art. For example, proteins may be enzymatically hydrolyzed, for example using a protease. For example, proteins may be hydrolyzed using alcalase (for example, at an enzyme:substrate ratio of about 1-15% by weight, for a duration of about 1-10 hours). The temperature may be in the range of about 40°C to 60°C, for example about 55°C. The reaction time may be, for example, 1-10 hours, and the pH value before starting the hydrolysis may be, for example, in the range of 6-9, preferably 6.5-8.5, more preferably 7.0-8.0.

[0098] The hydrolysis process may use porcine enzymes, in particular porcine pancreatic enzymes. For example, WO 9304593 A1 discloses a hydrolysis process using trypsin and chymotrypsin. The process includes a two-step hydrolysis reaction with a heat denaturation step in between to ensure that the final hydrolysate is substantially free of intact allergenic proteins. The trypsin and chymotrypsin used in these methods are preparations made from porcine pancreatic extracts.

[0099] WO2016156077A1 discloses a process for the preparation of milk protein hydrolysates, comprising hydrolyzing a milk-based proteinaceous material with a microbial alkaline serine protease in combination with bromelain, an Aspergillus derived protease, and a Bacillus derived protease.

[0100] Free Amino Acids The infant formula of the present invention may include free amino acids.

[0101] The levels of free amino acids may be selected to provide an amino acid profile sufficient for infant nutrition, in particular an amino acid profile that meets nutritional regulations (eg, European Commission Directive 2006 / 141 / EC).

[0102] For example, free amino acids can be incorporated into the eHF of the present invention to complement the amino acids contained in the peptide.

[0103] In the AAF, the protein content of infant formula is provided by free amino acids.

[0104] Examples of free amino acids for use in the infant formulas of the present invention include histidine, isoleucine, leucine, lysine, methionine, cysteine, phenylalanine, tyrosine, threonine, tryptophan, valine, alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, proline, serine, carnitine, taurine, and mixtures thereof.

[0105] Thus, suitably, the free amino acids in the eHF may be present at a concentration of 50% by weight or less, 40% by weight or less, 30% by weight or less, or 25% by weight or less, based on the total weight of the amino acids. Preferably, the eHF contains 25% by weight or less of free amino acids, based on the total weight of the amino acids. More preferably, the free amino acids in the eHF are present at a concentration of 20-25% by weight, 21-23% by weight, or about 22% by weight, based on the total weight of the amino acids.

[0106] The free amino acid content can be determined by any method known to those skilled in the art. Suitably, the free amino acid content can be obtained by separation of free amino groups present in an aqueous sample extract by ion exchange chromatography and optical detection after post-column derivatization with ninhydrin reagent. The total amino acid content can be obtained by hydrolysis of the test portion in 6 mol / L HCl under nitrogen and separation of the individual amino acids by ion exchange chromatography as described above.

[0107] In a preferred embodiment, the eHF of the present invention has no detectable peptides of a size greater than about 3000 Da; at least 95% by weight of the peptides have a molecular weight of less than about 1200 Da; optionally, at least about 45%, at least about 50%, or 45-55% by weight of the peptides have a molecular weight of 240-600 Da, and / or are dipeptides or tripeptides, and / or are 20-25%, 21-23%, or about 22% by weight based on the total weight of amino acids; the eHF of the present invention comprises about 1 g / L 2'-fucosyllactose and about 0.5 g / L lacto-N-neotetraose, and / or about 0.15 g / 100 kcal 2'-fucosyllactose and about 0.075 g / 100 kcal lacto-N-neotetraose; and the eHF does not contain added MCT.

[0108] carbohydrates The carbohydrate content of the infant formula of the present invention is preferably in the range of 9-14 g carbohydrate per 100 kcal.

[0109] The carbohydrate can be any carbohydrate suitable for use in infant formulas.

[0110] Examples of carbohydrates for use in the infant formula of the present invention include lactose, sucrose, maltodextrin, and starch. Mixtures of carbohydrates may also be used.

[0111] In one embodiment, the carbohydrate content comprises maltodextrin, hi one embodiment, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 50%, at least about 60%, or at least about 70% by weight of the total carbohydrate content is maltodextrin.

[0112] In one embodiment, the carbohydrate content comprises lactose, hi one embodiment, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 50%, at least about 60%, or at least about 70% by weight of the total carbohydrate content is lactose.

[0113] In one embodiment, the carbohydrates include lactose and maltodextrin.

[0114] Lipids The lipid content of the infant formula of the present invention is preferably in the range of 4.0-6.0 g lipid per 100 kcal.

[0115] The lipid can be any lipid or lipid suitable for use in infant formulas.

[0116] Examples of lipids for use in the infant formulas of the present invention include sunflower oil, low erucic acid rapeseed oil, safflower oil, canola oil, olive oil, coconut oil, palm kernel oil, soybean oil, fish oil, palm olein, high oleic sunflower oil, and high oleic safflower oil, and microbial fermentation oils containing long chain polyunsaturated fatty acids.

[0117] Lipids can also be in the form of oil-derived fractions such as palm olein, medium chain triglycerides (MCTs), and esters of fatty acids (e.g., arachidonic acid, linoleic acid, palmitic acid, stearic acid, docosahexaenoic acid, linoleic acid, oleic acid, lauric acid, capric acid, caprylic acid, caproic acid, etc.).

[0118] Further examples of lipids include structured lipids (i.e., lipids that have been chemically or enzymatically modified to change their structure). Preferably, the structured lipids are sn2 structured lipids, such as triglycerides that have an increased proportion of palmitic acid at the sn2 position of the triglyceride. Structured lipids may or may not be included.

[0119] Oils such as fish or microbial oils that are high in preformed arachidonic acid (ARA) and / or docosahexaenoic acid (DHA) may also be added.

[0120] Long chain polyunsaturated fatty acids such as dihomo-gamma-linolenic acid, arachidonic acid (ARA), eicosapentaenoic acid, and docosahexaenoic acid (DHA) may be added.

[0121] The infant formula may contain 2-20 mg ARA per 100 kcal, 5-15 mg ARA per 100 kcal, or about 10 mg ARA per 100 kcal, and / or 2-20 mg DHA per 100 kcal, 5-15 mg DHA per 100 kcal, or about 10 mg DHA per 100 kcal. Preferably, the infant formula contains about 10 mg ARA per 100 kcal and about 10 mg DHA per 100 kcal.

[0122] Medium Chain Triglycerides (MCT) High concentrations of MCTs may impair early weight gain. MCTs are not stored and do not support lipid accumulation. For example, Borschel et al. reported that infants fed a formula that did not contain MCTs gained significantly more weight between days 1 and 56 than infants fed a formula that contained 50% lipids derived from MCTs (Borschel, M., et al., 2018. Nutrients, 10(3), p. 289).

[0123] Thus, up to about 30% by weight of the lipids in the infant formulas of the present invention may be medium chain triglycerides (MCTs).

[0124] In some embodiments, about 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, 4% or less, 3% or less, 2% or less, 1% or less, 0.5% or less, or 0.1% or less by weight of the lipids are medium chain triglycerides (MCTs).

[0125] In some embodiments, 0-30%, 0-25%, 0-20%, 0-15%, 0-10%, 0-5%, 0-4%, 0-3%, 0-2%, 0-1%, 0-0.5%, or 0-0.1% by weight of the lipids are medium chain triglycerides (MCTs).

[0126] Preferably, the infant formula does not contain added MCTs. Preferably, about 0% by weight of the lipids are MCTs and / or the infant formula does not contain detectable MCTs. Preferably, the infant formula does not contain MCTs.

[0127] In preferred embodiments, the eHF of the present invention has no detectable peptides greater than about 3000 Da in size, and at least about 95% by weight of the peptides have a molecular weight of less than about 1200 Da; 45-55% by weight of the peptides have a molecular weight of 240-600 Da; the free amino acids are present at a concentration of 20-25% by weight based on the total weight of amino acids; and the eHF does not contain added MCT.

[0128] Further ingredients The infant formula of the present invention also preferably contains nutritionally significant amounts of all vitamins and minerals considered essential in the daily diet, with minimum requirements established for certain vitamins and minerals.

[0129] Examples of vitamins, minerals, and other nutrients for use in the infant formulas of the present invention include Vitamin A, Vitamin B1, Vitamin B2, Vitamin B6, Vitamin B12, Vitamin E, Vitamin K, Vitamin C, Vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in their salt form.

[0130] The infant formulas of the present invention may contain one or more carotenoids.

[0131] The infant formula of the present invention may also contain at least one probiotic. The term "probiotic" refers to a preparation of microbial cells or components of microbial cells that have a beneficial effect on the health or well-being of the host. Specifically, probiotics can improve intestinal barrier function.

[0132] A preferred probiotic is one that is generally safe, is an L(+) lactic acid producing culture, and has an acceptable shelf life for a product that is required to remain stable and effective for up to 24 months.

[0133] Examples of probiotic microorganisms for use in the infant formulas of the present invention include yeasts such as Saccharomyces, Debaromyces, Candida, Pichia, and Torulopsis, as well as species of the genera Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, and the like. Examples of yeasts that can be used include those of the genera Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus, and LactoBacillus.

[0134] Specific examples of suitable probiotic microorganisms are Saccharomyces cerevisiae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. casei, Lactobacillus casei shirota, Lactobacillus curvatus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei ... Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus salmonei, Lactobacillus lactis, Micrococcus varians, Pediococcus acidilactici, Pediococcus pentosus, Pediococcus acidilactici, Pediococcus halophilus, Streptococcus faecalis, Streptococcus thermophilus, Staphylococcus carnosis and Staphylococcus xylosus.

[0135] The infant formula of the present invention may also contain other substances that may have beneficial effects, such as prebiotics, lactoferrin, dietary fiber, nucleotides, nucleosides, etc.

[0136] Reducing, preventing, and treating the incidence of infectious diseases The infant formula of the present invention may be used to reduce the incidence of and / or prevent infections in infants.

[0137] The infant formula of the present invention may also be used to treat infections in infants.

[0138] As used herein, "reducing the incidence" of an infection means that the infant formula reduces the likelihood of an infection and / or at least one symptom associated with an infection.

[0139] As used herein, "preventing" an infectious disease means that the infant does not yet have the infectious disease and / or does not show any symptoms of the infectious disease, and that the infant formula prevents the infectious disease and / or prevents at least one symptom associated with the infectious disease.

[0140] As used herein, "treating" an infectious disease means that an infant has an infectious disease and the infant formula relieves, reduces, or ameliorates at least one symptom associated with the infectious disease and / or slows, reduces, or eliminates the infectious disease.

[0141] The term "infant" refers to a child under 12 months of age, for example, a child aged 0 to 6 months. Preferably, the infant has a cow's milk protein allergy.

[0142] In one aspect the invention provides a method of reducing, preventing or treating the incidence of an infectious disease in an infant comprising administering to the infant an infant formula according to the invention, in another aspect the invention provides an infant formula according to the invention for use in reducing, preventing or treating the incidence of an infectious disease in an infant.

[0143] As used herein, "infection" refers to the invasion of the infant's body tissues by any pathogen, including viruses, bacteria, fungi, parasites, and arthropods. Preferably, the infection is a bacterial or viral infection. The infection may be an ear infection, a respiratory infection, a gastrointestinal infection, and / or a urinary tract infection. Preferably, the infection is a respiratory infection and / or an ear infection. Most preferably, the infection is an ear infection.

[0144] The ear infection can be an external ear infection, a middle ear infection, and / or an inner ear infection. Preferably, the ear infection is a middle ear infection (otitis media). Otitis media (OM) is one of the most common childhood infections, the leading cause of medical visits in children, and the most common reason children receive antibiotics or undergo surgery in developed countries. Streptococcus pneumoniae, Moraxella catarrhalis, and non-encapsulated Haemophilus influenzae are the main bacterial pathogens causing OM. Viruses, alone or together with bacterial co-pathogens, are responsible for the increase in OM. Although many respiratory viruses contribute to viral OM, it is most frequently caused by respiratory syncytial virus or rhinovirus (Rovers, MM, et al., 2004. Otitis media. The lancet, 363(9407), pp. 465-473).

[0145] Ear infections can be acute OM (AOM) or OM with effusion (OME), which are different stages of the OM continuum. AOM is defined as the presence of middle ear effusion (MEE) in conjunction with the rapid onset of one or more signs or symptoms of inflammation in the middle ear, such as ear pain or pulling, otorrhea, fever, or irritability. OME is defined as MEE without signs or symptoms of acute infection. Some children present with non-infectious discomfort, including hearing loss, irritability, impaired dexterity, or sleep disorders (Rovers, MM, et al., 2004. Otitis media. The lancet, 363(9407), pp.465-473).

[0146] Accordingly, the infant formula of the present invention can be used to reduce, prevent, or treat the occurrence of AOM. In another embodiment, the formula of the present invention can be used to reduce, prevent, or treat the occurrence of OME. In another embodiment, the formula of the present invention can be used to reduce, prevent, or treat the occurrence of MEE, earache or ear tugging, ear discharge, fever, irritability, hearing loss, fine motor impairment, or sleep disorder.

[0147] In some embodiments, the infectious disease is a respiratory infection. The disease burden due to respiratory infections is considered to be greater than that of any other disease cause. In 2002, 18% of the mortality rate of children under 5 years old was due to respiratory infections. Respiratory infections can be upper respiratory tract infections (URTIs) and / or lower respiratory tract infections (LRTIs) (Tregoning, J.S. and Schwarze, J., 2010. Clinical microbiology reviews, 23(1), pp. 74-98).

[0148] URTIs can be characterized by the following clinical symptoms: nose (runny nose, nasal congestion, nose picking, yellow discharge, bloody discharge, sneezing); and / or pharynx (itching of the pharynx, sore throat, hoarseness); accompanied by fever (above 38°C) and / or otitis media.

[0149] LRTIs can be characterized by the following clinical symptoms: pneumonia (an episode of pneumonia can be defined as a disease reported as cough or dyspnea); and / or bronchitis (cough and / or secretion and / or yellow secretion).

[0150] Accordingly, the infant formula of the present invention can be used to reduce, prevent, or treat the occurrence of one or more of URTI and / or LRTI infections, and / or runny nose, nasal congestion, nose picking, yellow discharge, bloody discharge, sneezing, itching of the pharynx, sore throat, hoarseness, fever, otitis media, pneumonia, or bronchitis.

[0151] The infant formulas of the present invention may reduce the risk of an infant developing an infection as described herein, for example, LTRI, UTRI, and / or OM.

[0152] Reducing the incidence of infectious diseases in infants may mean that the risk of an infant developing an infectious disease is reduced when administered an infant formula according to the invention compared to when administered a conventional infant formula, such as the control formula used in the examples.

[0153] In one embodiment, the risk of infection may be reduced from when the infant formula is first administered to the infant (V0) to 6 months after the infant formula is first administered to the infant (V6), preferably from V0 to 4 months after the infant formula is first administered to the infant (V4), for example, when the infant formula has been administered for 4 months or more. The risk of infection may also be reduced from V4 to V6.

[0154] The infant formula of the present invention may reduce the use of antipyretics and / or antibiotics as concomitant medications (e.g., during and / or after the administration period of the infant formula). As used herein, an "antipyretic" is a substance known to those skilled in the art that reduces fever. As used herein, an "antibiotic" is a substance known to those skilled in the art that is used to treat or prevent some types of bacterial infections.

[0155] The inventors have surprisingly shown that the infant formula of the present invention reduces the amount of concomitant medication.For example, the infant formula of the present invention can reduce the amount of antipyretics used as concomitant medication.The infant formula of the present invention can also reduce the amount of antibiotics used as concomitant medication.

[0156] Thus, in another aspect, the present invention provides a method of reducing the amount of antipyretic medication used by an infant, the method comprising administering to the infant an infant formula according to the present invention.In another aspect, the present invention provides an infant formula according to the present invention for use in reducing the amount of antipyretic medication used by an infant.

[0157] Use of an infant formula according to the invention preferably reduces the amount of medication (eg antipyretics) required compared to use of a conventional infant formula, such as the control formula used in the examples.

[0158] Cow's milk protein allergy and obesity In addition to preventing, reducing the incidence of, or treating infectious diseases, the infant formulas of the present invention may prevent obesity in infants and / or reduce the risk of obesity in infants and / or prevent cow's milk protein allergies in infants.

[0159] Thus, in a preferred aspect, the present invention provides an infant formula according to the present invention for use both in (i) preventing, reducing the incidence or treating infectious diseases in infants and (ii) preventing / treating obesity in infants and / or reducing the risk of obesity in infants. Further, the infant formula may be used in the treatment and / or prevention of cow's milk protein allergy (CMPA) and / or one or more symptoms of CMPA.

[0160] Most children with CMP allergy (CMPA) have two or more symptoms: 50-70% have skin symptoms; 50-60% have gastrointestinal symptoms; and 20-30% have respiratory tract symptoms. In 10% of children, severe, life-threatening symptoms may develop. (Nutten, 2018. EMJ Allergy Immunol, 3(1), pp.50-59).

[0161] Cutaneous symptoms may include urticaria, atopic eczema, and angioedema. Gastrointestinal symptoms may include dysphagia, frequent regurgitation, colic, abdominal pain, vomiting, anorexia, anorexia, diarrhea (with or without intestinal protein loss or bleeding), constipation (with or without perianal rash), failure to thrive, occult blood loss, and iron deficiency anemia. Respiratory symptoms may include runny nose, wheezing, and chronic cough. Other systemic symptoms include anaphylaxis and shock-like symptoms with severe metabolic acidosis, vomiting, and diarrhea (food protein-induced gastroenteritis). These symptoms are typically unrelated to infection, drug intake, or other causes (Koletzko, S., et al., 2012. Journal of pediatric gastroenterology and nutrition, 55(2), pp. 221-229).

[0162] Infants with CMPA may require higher levels of protein due to, for example, gastrointestinal symptoms. Thus, eHF typically contains 2.6-2.8 g protein per 100 kcal, and AAF typically contains 2.8-3.1 g protein per 100 kcal, to accommodate the needs of infants with gastrointestinal pathology, including severe malabsorption, or who require more protein and calories to compensate for a high metabolic rate.

[0163] However, consumption of high-protein infant formulas has been associated with increased body weight and body mass index at age 2 years, as well as increased plasma essential amino acids, insulin-like growth factor-1, and C-peptide concentrations, which may induce weight gain and lipogenic activity. Lower protein content may reduce the risk of subsequent obesity (Totzauer, M., et al., 2018. Obesity, 26(7), pp. 1203-1210).

[0164] For example, the European Childhood Obesity Project (CHOP) has shown that infants fed infant formulas containing high protein levels gained more weight during the first year of life and had a higher BMI and risk of obesity at age 6 than infants fed infant formulas containing lower protein levels (Totzauer, M., et al., 2018. Obesity, 26(7), pp.1203-1210).

[0165] The inventors have surprisingly shown that the infant formula of the present invention supports proper growth and development in allergic infants. Moreover, the infant formula is safe and well tolerated.

[0166] Manufacturing method The infant formulas of the present invention may be prepared in any suitable manner.

[0167] For example, the infant formula can be prepared by blending together a hydrolyzed protein source, a carbohydrate source, and a lipid source in the appropriate ratio. If an emulsifier is used, it can be included at this point. Vitamins and minerals can be added at this point, but vitamins are usually added later to avoid thermal degradation. Any lipophilic vitamins and emulsifiers, etc. can be dissolved in the lipid source before blending. Water, preferably reverse osmosis treated water, can then be mixed to form a liquid mixture. A commercially available liquefaction device can be used to form the liquid mixture. The liquid mixture can then be homogenized.

[0168] The liquid mixture can then be heat treated to reduce the bacterial content, for example by steam injection or using an autoclave or a heat exchanger, for example a plate heat exchanger.

[0169] The liquid mixture may then be cooled and / or homogenized. The pH and solids content of the homogenized mixture may be adjusted at this point.

[0170] The homogenized mixture can then be transferred to suitable drying equipment, such as a spray dryer or freeze dryer, and converted to a powder. If a liquid infant formula is desired, the homogenized mixture may be sterilized and then aseptically filled into suitable containers, or it may be filled into containers first and then retorted. EXAMPLES

[0171] The present invention will now be further described by way of examples, which are meant to be provided to aid those skilled in the art in practicing the invention, and are not intended to limit the scope of the invention in any way.

[0172] Example 1 - Exemplary Highly Hydrolyzed Infant Formula The following is an example of an extensively hydrolyzed infant formula according to the present invention: The eHF of the present invention preferably contains nutritionally significant amounts of all nutrients, vitamins, and minerals considered essential in the daily diet. Minimum requirements have been established for certain nutrients, vitamins, and minerals.

[0173] [Table 1]

[0174] Example 2 - Safety and efficacy of highly hydrolyzed infant formula with reduced protein content.

[0175] Study design The safety and efficacy of a reduced-protein, extensively hydrolyzed infant formula was investigated in a controlled, double-blind, randomized, multicenter, interventional clinical trial with two parallel groups fed the formula.

[0176] The aim of the clinical trial was to demonstrate that infants with cow's milk protein allergy (CMPA) fed a novel eHF with reduced protein levels (Althera 2.2) supplemented with two human milk oligosaccharides (HMOs) (test formula) performed comparably to infants fed a commercial eHF without HMOs (Althera 2.5) (control formula). Commercial eHF is currently approved as a food for specific medical purposes (Regulation (EU) 2016 / 128). The primary outcome of the study was daily weight gain from enrolment to the 4-month follow-up, with the age window at enrolment being 0-6 months (non-inferiority design).

[0177] Other objectives included assessing whether CMPA infants receiving the study formula (i) reduced medication use and risk / morbidity of infections, including ear infections and lower respiratory tract infections, (ii) was tolerable and enabled age-appropriate growth, and (iii) reduced healthcare costs. Thus, secondary endpoints of the study were: (1) changes in weight-for-age, length-for-age, and other growth parameters, including head circumference for age Z-scores (WHO growth standards); (2) safety (incidence of adverse events and serious adverse effects); and (3) events of interest (incidence of infections, medication use).

[0178] The study population was full-term infants with physician-diagnosed CMPA according to standard clinical practice and with at least two symptoms per inclusion criteria. One hundred and thirty infants were required to complete four months of study formula intake.

[0179] The selection criteria were as follows: 1. Term infants (37 weeks ≤ gestational age ≤ 42 weeks); 2.2500g≦birth weight≦4500g; 3. Written informed consent from the infant's parent (or parents, if required by national regulations) or legally authorized representative (LAR); 4. Infants up to 6 months of age; 5. Those who were exclusively formula-fed at the time of enrollment, or mothers of breast-fed CMPA infants who voluntarily elected to exclusively formula-fed prior to enrollment: and 6. Infants diagnosed with CMPA by a physician according to standard clinical practice (and not being treated with extensively hydrolyzed milk or amino acid infant formula) who have at least two of the following symptoms present - crying, regurgitation, liquid stools or constipation, atopic skin lesions, urticaria, or respiratory symptoms. Only one of the above symptoms need be present if the diagnosis is based on either a positive IgE blood test, skin prick test, patch test, or food challenge test.

[0180] The exclusion criteria were as follows: 1. Congenital diseases or abnormalities that may affect development. 2.Demonstrated chronic malabsorption not due to CMPA. 3. Significant prenatal and / or postnatal serious illness other than CMPA prior to enrollment (as per the investigator's medical judgment). 4. Parents of a minor. 5. Infants whose parents or caregivers cannot be expected to comply with study procedures. 6.Currently participating in another clinical trial or have participated in one since birth.

[0181] The test and control formulas are shown below.

[0182] [Table 2]

[0183] In the test and control formulas, More than 99% by weight of the peptides had a molecular weight less than 3000 Da. More than 95% by weight of the peptides had a molecular weight less than 1200 Da. Approximately 52% by weight of the peptides were dipeptides and tripeptides (peptides with molecular weights of 600-240 Da). Approximately 22.4% by weight of the total amino acids were free amino acids in the test and control formulas. There was no detectable beta-lactoglobulin (i.e. the beta-lactoglobulin content was less than 0.01mg / kg). There was no detectable casein (i.e. the casein content was less than 0.2mg / kg).

[0184] Both formulas were in powder form and were prepared for oral intake by the infant in amounts appropriate for their weight, age, and appetite, according to the instructions printed on the product label on the can.

[0185] Infants received the study formula for a minimum of 4 months after baseline (main study period) and for as long as the infant required according to medical prescription (up to 12 months of age).

[0186] The daily intake required by infants varied according to age, weight and appetite. The products were given to infants ad libitum but parents or caregivers followed guidelines printed on the label and / or were advised by study personnel regarding the appropriate amount to be given per day.

[0187] Infants will have up to seven study visits: baseline (at enrollment), monthly (+1, +2, +3, +4 months) until 4 months after baseline, and 6 months after baseline. One additional final visit is planned when the infants reach 12 months of age.

[0188] Randomization was 1:1 per study formula group and was performed by minimization in Medidata Balance. Stratification was by age at enrollment (0-60 days, 61-120 days, >120 days), sex, and mode of delivery (vaginal or caesarean section). Twins enrolled were randomized to the same formula.

[0189] Test results The study showed that the test formula supported proper growth and development in allergic infants and was safe and well tolerated.

[0190] The primary analysis assessed whether growth was noninferior in infants fed the test formula compared with infants fed the control formula over the first 4 months of treatment. Treatment differences and one-sided simultaneous 97.5% confidence intervals were performed with mixed models. Weight (kg) was modeled as a function of age (months), treatment, sex, age*treatment, and age*sex. The models assumed a variance-covariance matrix with an autocorrelated type I structure for outcomes at adjacent visits. The calculated differences were used to estimate the treatment effect from day 63 [(14+112) / 2]. Estimates were converted from kg / month to g / day using a divisor of 0.0305. Key secondary analyses included weight, height, and head circumference, stool consistency, behavioral patterns, and health resource utilization.

[0191] Infants fed the test formula with reduced protein and HMO achieved normal growth according to WHO growth standards. Specifically, the primary analysis showed that weight gain [g / d] of infants fed the test formula was non-inferior to growth in those fed the control formula (Figure 1). There were no significant differences in any of the anthropometric parameters at any time up to the 4-month follow-up.

[0192] The safety profiles of the test and control formulas were similar. The incidence of otitis media / middle ear infection was significantly lower in the groups fed the test formula between V0–V4 and V0–V6 (Figure 3).

[0193] Non-significant relative risk reductions were also seen for lower and upper respiratory tract infections (Figure 3). Use of antipyretics was significantly lower in the test formula groups V4-V6 (Figure 4).

[0194] Example 3 - Exemplary Amino Acid-Based Infant Formula The following is an example of an amino acid-based infant formula according to the present invention. The AAF of the present invention preferably contains nutritionally significant amounts of all nutrients, vitamins, and minerals considered essential in the daily diet. Minimum requirements have been established for certain nutrients, vitamins, and minerals.

[0195] [Table 3]

[0196] All publications mentioned in the above specification are incorporated herein by reference. Various modifications and alterations of the disclosed methods, cells, compositions, and uses of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been disclosed in connection with certain preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the disclosed modes for carrying out the invention, which are obvious to those skilled in the art, are intended to be within the scope of the following claims.

Claims

1. 1. An infant formula for use in reducing the incidence of infections in infants or in preventing infections in infants, said infant formula being an extensively hydrolyzed infant formula (eHF) or an amino acid based infant formula (AAF), said infant formula comprising protein, carbohydrate and lipid, said eHF comprising 1.8-2.4g protein per 100 kcal or said AAF comprising 1.8-2.9g protein per 100 kcal, said infant formula further comprising 2'-fucosyllactose (2'FL) and / or lacto-N-neotetraose (LNnT), and 0-30% by weight of the lipid is medium chain triglycerides (MCT); An infant formula, wherein the infant has a cow's milk protein allergy.

2. 2. An infant formula for use according to claim 1, wherein said infant formula comprises 2'FL and LNnT.

3. 3. An infant formula for use according to claim 1 or 2, wherein said infant formula comprises 0.5-3 g / L, 0.8-1.5 g / L, or 1 g / L of 2'FL.

4. An infant formula for use as described in claim 3, wherein the infant formula contains 1 g / L of 2'FL.

5. 5. An infant formula for use according to any one of claims 1 to 4, wherein said infant formula comprises 0.2-1 g / L, 0.5-0.8 g / L, or 0.5 g / L of LNnT.

6. An infant formula for use as described in claim 5, wherein the infant formula contains 0.5 g / L LNnT.

7. An infant formula for use according to any one of claims 1 to 6, wherein said infant formula comprises 1 g / L 2'FL and 0.5 g / L LNnT.

8. An infant formula for use according to any one of claims 1 to 7, wherein said infant formula is eHF and said infant formula comprises 2.0 to 2.4 g protein per 100 kcal.

9. An infant formula for use as described in any one of claims 1 to 7, wherein the infant formula is eHF and contains 2.1 to 2.3 g of protein per 100 kcal.

10. An infant formula for use as described in any one of claims 1 to 7, wherein the infant formula is eHF and contains 2.15 to 2.25 g of protein per 100 kcal.

11. An infant formula for use according to any one of claims 1 to 7, wherein said infant formula is AAF and said infant formula contains 1.9 to 2.8 g protein per 100 kcal.

12. An infant formula for use as described in any one of claims 1 to 7, wherein the infant formula is AAF and contains 2.0 to 2.7 g of protein per 100 kcal.

13. An infant formula for use as described in any one of claims 1 to 7, wherein the infant formula is AAF and contains 2.0 to 2.6 g or 2.0 to 2.4 g of protein per 100 kcal.

14. An infant formula for use according to any one of claims 1 to 13, wherein said infant formula comprises 2.2 g protein per 100 kcal.

15. 15. An infant formula for use according to any one of claims 1 to 14, wherein less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, or less than 1% by weight of the lipids in the infant formula are medium chain triglycerides (MCT).

16. An infant formula for use according to any one of claims 1 to 15, wherein said infant formula does not contain added MCTs.

17. Infant formula for use according to any one of claims 1 to 16, wherein said infant formula comprises 9 to 14 g carbohydrates per 100 kcal and / or 4.0 to 6.0 g lipids per 100 kcal.

18. Infant formula for use according to any one of claims 1 to 17, wherein the infection is an ear infection, a respiratory infection, a gastrointestinal infection and / or a urinary tract infection.

19. Infant formula for use according to any one of claims 1 to 18, wherein the infection is a respiratory infection.

20. An infant formula for use according to any one of claims 1 to 18, wherein the infection is an upper respiratory tract infection and / or a lower respiratory tract infection.

21. An infant formula for use according to any one of claims 1 to 18, wherein the infection is an ear infection.

22. An infant formula for use according to any one of claims 1 to 18, wherein the infection is a middle ear infection.

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