WO2023159093A1

WO2023159093A1 - Nutritional compositions comprising human milk oligosaccharides and bovine immunoglobulin

Info

Publication number: WO2023159093A1
Application number: PCT/US2023/062697
Authority: WO
Inventors: Rachael Buck; Michael Farrow; David Daly; Sinead MORRIN; Kevin Murphy
Original assignee: Abbott Laboratories
Priority date: 2022-02-16
Filing date: 2023-02-16
Publication date: 2023-08-24

Abstract

Disclosed are methods for treating or preventing infections in an individual and nutritional compositions for administering to individuals for the purpose of treating and/or preventing infections. The nutritional compositions include a combination of a human milk oligosaccharide and a protein ingredient having an enriched amount of one or more bovine immunoglobulin(s).

Description

NUTRITIONAL COMPOSITIONS COMPRISING HUMAN MILK OLIGOSACCHARIDES AND BOVINE IMMUNOGLOBULIN

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63/310,919, filed February 16, 2022, the entire content of which is incorporated herein by reference.

FIELD

[0002] The present disclosure relates to a blend of human milk oligosaccharides (HMOs) and a bovine milk component for nutritionally benefitting an individual consumer, including improving one or more aspects of the individual’s health. More particularly, the present disclosure relates to nutritional compositions comprising a blend of HMOs and bovine immunoglobulin that can modulate or improve the health of an individual consumer by treating or preventing certain infections or by reducing or preventing one or more symptoms of an infection as described in greater detail herein.

BACKGROUND

[0003] Breast milk containing Human Milk Oligosaccharides has been associated with enhanced development and balanced growth and maturation of infant's respiratory, gastrointestinal, and immune systems, among others, thereby providing protection to the infant from a variety of diseases. There are more than 200 known oligosaccharides present in human milk, with various individual classes having been shown to promote health in a variety of ways. It appears that certain infections are prevented by HMOs due to their activity as pathogen receptor analogues (i.e., receptor mimics that prevent pathogenic adhesion). Further, breast milk includes HMOs that not only act as pathogen receptor analogues, but certain structures activate immune factors as well as acting as metabolic fuel in the gut microbiome to preferentially benefit certain beneficial bacteria.

[0004] Due to the accepted safety of HMOs as a component of an infant’s diet, efforts have been extended to develop the use of HMOs in other therapeutic regimes. However, these efforts have yet to develop fully effective therapies in many areas. Accordingly, there is an unmet need for nutritional solutions that are effective to treat one or more of the conditions or diseases described herein, including but not, limited to treatment of respiratory infections.

SUMMARY

[0005] Respiratory infection and related pathology is a leading cause of hospitalizations in children under 5. Likewise, such infections also are of great impact on the elderly and otherwise immunocompromised individuals. Vaccines and therapies for many respiratory and gastrointestinal viruses remain elusive. Treatments for many of these viruses rely on a rapid pharmaceutical response to established infections in order to alleviate symptoms. The current invention demonstrates a path towards a nutritional intervention for treating, preventing, or reducing the extent to which respiratory viruses can establish sites of infection. By reducing and/or preventing a virus’s ability to proliferate, the present invention reduces the necessity of using drug treatments to reduce illness symptoms.

[0006] By providing immunoglobulins and HMO(s) in products for daily consumption, the present invention would provide a passive immune defense against highly transmissible viral infections that affect infants, the elderly, and immunocompromised individuals. The general inventive concepts are based, in part, on the discovery of an unexpected synergy between bovine immunoglobulin-containing protein components and HMOs at reducing pathogenic infectivity.

[0007] The general inventive concepts are directed to nutritional compositions including at least one HMO, in combination with at least one bovine immunoglobulin-containing protein component, for use in modulating or treating the conditions or diseases described herein, including but not limited to: supplementing an immune response in an individual; treating an infection in an individual; preventing an infection in an individual; and reducing or ameliorating symptoms of an infection in an individual.

[0008] In certain exemplary aspects, the general inventive concepts relate to a method of reducing or ameliorating symptoms of a viral infection comprising administering a nutritional composition comprising a bovine immunoglobulin-containing protein component and a human milk oligosaccharide component to the individual, wherein the human milk oligosaccharide (HMO) component comprises at least one neutral HMO and at least one acidic HMO. [0009] In certain exemplary aspects, the general inventive concepts relate to a method of supplementing an immune response to a viral infection comprising administering a nutritional composition comprising a bovine immunoglobulin-containing protein component and a human milk oligosaccharide component to the individual, wherein the human milk oligosaccharide (HMO) component comprises at least one neutral HMO and at least one acidic HMO.

[0010] In certain exemplary aspects, the general inventive concepts relate to a method of treating or preventing a viral infection comprising administering a nutritional composition comprising a bovine immunoglobulin-containing protein component and a human milk oligosaccharide component to the individual, wherein the human milk oligosaccharide (HMO) component comprises at least one neutral HMO and at least one acidic HMO.

[0011] In certain exemplary aspects, the general inventive concepts relate to a method of preparing a bovine immunoglobulin-containing protein component, comprising: subjecting a milk fraction comprising skim milk and/or whey to microfiltration to defat and decaseinate the milk fraction and provide a sterile retentate, subjecting the defatted and decaseinated milk fraction to a first ultrafiltration step and a volume concentration factor (VCF) of at least about 5, and subjecting the concentrated milk fraction to a second ultrafiltration step to remove lactose and mineral content.

[0012] In certain exemplary aspects, the general inventive concepts relate to a nutritional composition for at least one of: supplementing an immune response in an individual; treating an infection in an individual; preventing an infection in an individual; and/or reducing or ameliorating symptoms of an infection in an individual, the nutritional composition comprising from about 0.01 mg/mL to about 100 mg/mL of a bovine immunoglobulin-containing protein component; from about 0.01 mg/mL to about 20 mg/mL of a human milk oligosaccharide component comprising at least one neutral HMO and at least one acidic HMO. In certain instances, the composition also comprises from about 0.001 g/L to about 1 g/L of a long chain polyunsaturated fatty acid component comprising docosahexaenoic acid and arachidonic acid; from about 0.001 pg/mL to about 5 pg/mL of a carotenoid component; from about 10 mg/L to about 300 mg/L of a monomeric monophosphate nucleotide component; and wherein the nutritional composition is shelf stable for at least 3 months. In certain aspects, the aforementioned methods comprise administration of the nutritional composition.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 shows a bar graph of influenza A viral infectivity in the presence of a number of compositions.

[0014] Figure 2 shows a bar graph of respiratory syncytial virus (RSV) infectivity in the presence of a variety of immunoglobulin-containing compositions.

DETAILED DESCRIPTION

[0015] The nutritional compositions and methods described herein utilize a mixture of HMOs (e.g., neutral and acidic HMOs) and bovine immunoglobulin-containing protein component alone or in combination with long chain polyunsaturated fatty acids, antioxidants, and also including carotenoids, nucleotides, or both, for controlling, preventing, and/or treating a number of diseases and conditions as described in greater detail herein. These and other features of the nutritional compositions and methods, as well as some of the many optional variations and additions, are described in detail hereafter.

[0016] The term “human milk oligosaccharide” or “HMO”, as used herein, unless otherwise specified, refers generally to a number of complex carbohydrates found in human breast milk that can be in acidic or neutral form, and, in certain instances, to precursors thereof. Exemplary non-limiting human milk oligosaccharides include neutral oligosaccharides (which include fucosylated oligosaccharides and N-acetylated oligosaccharides (e.g., neutral non- fucosylated)) and acidic oligosaccharides (which include sialylated oligosaccharides). Specific non-limiting embodiments include: 2'-fucosyllactose (2’-FL), 3-fucosyllactose (3-FL), 3'- sialyllactose (3’-SL), 6'-sialyllactose (6’-SL), lacto-N-tetraose (LNT), and lacto-N-neotetraose (LNnT). The term “HMO blend,” unless specified otherwise, refers to a combination of neutral and acidic HMOs, which may comprise at least two of 2’ -FL, 3-FL, 3’-SL, 6’-SL, and LNT.

[0017] The term “bovine immunoglobulin-containing protein component”, as used herein, refers to a protein source (often a milk protein fraction, e.g., whey) that has been subjected to a process designed to enrich the amount of immunoglobulin relative to native or supplemented (milk) protein sources. The term does not refer to protein sources supplemented with immunoglobulin. In certain aspects, the process involves removing or reducing the amount of at least one of casein, fat, bacteria, lactose, minerals, bovine serum albumin and P- lactoglobulin. In certain aspects, the process involves removing or reducing the amount of each of casein, fat, bacteria, lactose, minerals, bovine serum albumin and P-lactoglobulin. In certain aspects, the term refers to a protein component wherein the ratio of native immunoglobulins is enriched, e.g., to increase the relative amount of one over others. In certain aspects, the term refers to a protein source that includes an enriched amount of IgA, including an enriched ratio of IgA relative to IgG.

[0018] The terms “fat” and “oil” as used herein, unless otherwise specified, are used interchangeably to refer to lipid materials derived or processed from plants or animals. These terms also include synthetic lipid materials so long as such synthetic materials are suitable for oral administration to humans.

[0019] The term “shelf stable” as used herein, unless otherwise specified, refers to a nutritional composition that remains commercially stable after being packaged and then stored at 18-24° C. for at least 3 months, including from about 3 months to about 36 months, including from about 3 months to about 24 months, and also including from about 3 months to about 18 months.

[0020] The terms “nutritional formulation” or “nutritional composition” as used herein, are used interchangeably and, unless otherwise specified, refer to synthetic formulas including nutritional liquids, nutritional powders, nutritional solids, nutritional semi-solids, nutritional semi-liquids, nutritional supplements, and any other nutritional food product as known in the art. The nutritional powders may be reconstituted to form a nutritional liquid, all of which comprise one or more of fat, protein and carbohydrate and are suitable for oral consumption by a human. The terms “nutritional formulation” or “nutritional composition” do not include human breast milk and do not refer to supplemented milk, whether of human origin or otherwise. [0021] The term “nutritional liquid” as used herein, unless otherwise specified, refers to nutritional compositions in ready-to-drink liquid form, concentrated form, and nutritional liquids made by reconstituting the nutritional powders described herein prior to use.

[0022] The term “nutritional powder” as used herein, unless otherwise specified, refers to nutritional compositions in flowable or scoopable form that can be reconstituted with water or another aqueous liquid prior to consumption and includes both spraydried and drymixed/dryblended powders.

[0023] The term “nutritional semi-solid,” as used herein, unless otherwise specified, refers to nutritional compositions that are intermediate in properties, such as rigidity, between solids and liquids. Some semi-solids examples include puddings, gelatins, and doughs.

[0024] The term “nutritional semi-liquid,” as used herein, unless otherwise specified, refers to nutritional compositions that are intermediate in properties, such as flow properties, between liquids and solids. Some semi-liquids examples include thick shakes and liquid gels.

[0025] The term “individual” as used herein, refers generally to a preterm infant, infant, toddler, child, or adult, including elderly adults. In certain instances, the individual is immune compromised.

[0026] The term “infant” as used herein, refers generally to individuals up to age 36 months of age, actual or corrected.

[0027] The term “preterm infant” as used herein, refers to those infants bom at less than 37 weeks gestation, have a birth weight of less than 2500 gm, or both.

[0028] The term “reconstitute” or various other forms such as “reconstituted” or “reconstituting” all refer to the general act of adding a suitable amount of liquid, typically water, to a form of nutritional formulation that is not in its ready-to-drink liquid form, such as nutritional powder or a concentrated form of a nutritional liquid, thereby making the nutritional composition ready-to-drink. [0029] The terms “susceptible” and “at risk” as used herein, unless otherwise specified, mean having little resistance to a certain condition or disease relative to the general population (e.g., immunocompromised individuals), including being genetically predisposed, having a family history of, and/or having symptoms of the condition or disease. The term refers to those having a vulnerability higher than the general population. In certain instances, the individual is need of a therapeutic intervention, e.g., is immune compromised.

[0030] The terms “modulating” or “modulation” or “modulate” as used herein, unless otherwise specified, refer to the targeted movement of a selected characteristic.

[0031] The term “ameliorate” as used herein, unless otherwise specified, means to eliminate, delay, or reduce the prevalence or severity of symptoms associated with a condition or disease.

[0032] The term “an effective amount” is intended to qualify the amount of HMO and/or bovine immunoglobulin-containing protein component which will achieve the goal of preventing or treating a disease or that which will achieve the goal of decreasing the risk that the patient will suffer an adverse health event, including reducing or preventing one or more symptoms, while avoiding adverse side effects such as those typically associated with alternative therapies. The effective amount may be administered in one or more doses.

[0033] The terms “treating” and “treatment” as used herein, unless otherwise specified, includes delaying the onset of a condition, reducing the severity of symptoms of a condition, or eliminating some or all of the symptoms of a condition. In certain aspects, symptoms of a viral infection include, but are not limited to, at least one of headache, chills, coughing, fussiness and irritability, runny nose, red eyes, otitis media, wheezing, brief apneas, fever, fatigue, decreased appetite, dysphagia, pharyngitis, bronchitis, and pneumonia. In certain aspects, treatment or prevention of a viral infection includes a reduction in viral load or a reduction in viral replication.

[0034] The term “food products” as used herein refer to delivery vehicles that contain one or more of fats, amino nitrogen and carbohydrates and provides some or all of the nutritional support for a patient in the recommended daily amounts. Frequently a food product will contain vitamins, minerals, trace minerals and the like to provide balanced nutrition to meal replacements, medical foods, supplements. The food products may be in any typical form such as beverages, powders, bars, juices, carbonated beverages, bottled water, etc.

[0035] As used herein, all concentrations expressed as either “pg/liter,” “mg/liter,” “mg/L,” “mcg/L,” “mg/mL” etc., refer to ingredient concentrations within the described nutritional compositions as calculated on an as-fed basis (e.g., reconstituted for consumption in the case of nutritional powders), unless otherwise specified.

[0036] All percentages, parts and ratios as used herein, are by weight of the total composition, unless otherwise specified. All such weights, as they pertain to listed ingredients, are based on the active level and, therefore, do not include solvents or by-products that may be included in commercially available materials, unless otherwise specified.

[0037] Numerical ranges as used herein are intended to include every number and subset of numbers within that range, whether specifically disclosed or not. Further, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 2 to 8, from 3 to 7, from 5 to 6, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

[0038] All references to singular characteristics or limitations of the general inventive concepts shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made.

[0039] All combinations of method or process steps as used herein can be performed in any order, unless otherwise specified or clearly implied to the contrary by the context in which the referenced combination is made.

[0040] The HMOs and bovine immunoglobulin-containing protein component can be formulated in suitable compositions and then, in accordance with the methods of the invention, administered to an individual in a form adapted to the chosen route of administration. The formulations include, but are not limited to, those suitable for oral or parental (including subcutaneous, intramuscular, intraperitoneal, intratumoral, and intravenous) administration. Oral administration, as defined herein, includes any form of administration in which the HMOs and immunoglobulin pass through the esophagus of the patient. For example, oral administration includes nasogastric intubation, in which a tube is run from through the nose to the stomach of the patient to administer food or drugs.

[0041] Oral formulations include any solid, liquid, or powder formulation suitable for use herein, provided that such a formulation allows for the safe and effective oral delivery of the HMOs, immunoglobulin, and optional nutritive components. In preferred aspects, the oral formulation is a liquid nutritional composition. Formulations of the present invention suitable for oral administration may be presented as discrete units such as tablets, troches, capsules, lozenges, wafers, or cachets, each containing a predetermined amount of the HMO and immunoglobulin as a powder or granules or as a solution or suspension in an aqueous liquor or non-aqueous liquid such as a syrup, an elixir, an emulsion, or a draught.

[0042] The nutritional compositions and methods may comprise, consist of, or consist essentially of the essential elements of the compositions and methods as described herein, as well as any additional or optional element described herein or otherwise useful in nutritional composition applications.

Human Milk Oligosaccharides

[0043] The nutritional compositions according to the general inventive concepts include an HMO component comprising at least one HMO, and in many aspects, a combination of two or more HMOs, including neutral HMOs and acidic HMOs. While N-acetylated oligosaccharides are described in greater detail apart from e.g., fucosylated oligosaccharides, those of ordinary skill in the art will recognize that the term “neutral HMO” is understood to include both fucosylated HMOs and N-acetylated HMOs, unless the context indicates to the contrary.

[0044] The HMOs may be included in the nutritional compositions with a bovine immunoglobulin-containing protein component alone, or in some aspects, in combination with other functional components (LCPUFAs, antioxidants, nucleotides, etc.) to aid in ameliorating symptoms of the particular condition or disease as described herein. The HMO or HMOs may be isolated or enriched from milk(s) secreted by mammals including, but not limited to: human, bovine, ovine, porcine, or caprine species. The HMOs may also be produced via microbial fermentation, enzymatic processes, chemical synthesis, or combinations thereof.

[0045] Suitable HMOs for use in the nutritional compositions and HMO components may include neutral oligosaccharides (including neutral, non-fucosylated oligosaccharides), acidic HMOs, and n-acetyl glucosylated oligosaccharides (a subset of neutral oligosaccharides that are generally characterized as non-fucosylated and non-sialylated oligosaccharides). In certain aspects, the nutritional composition may also include HMO precursors. Specific nonlimiting examples of HMOs that may be included individually or in combination in the compositions according to the general inventive concepts include: 2’-FL, 3-FL, 3’-SL, 6'-SL, LNnT, and LNT.

[0046] Other HMOs that may be included in certain embodiments include: N- acetylglucosamine (GlcNAc); L-fucose (L-Fuc); D-fucose (D-Fuc); fucosyl oligosaccharides (z.e., Lacto-N-fucopentaose I; Lacto-N-fucopentaose II; Lacto-N-fucopentaose III; Lacto-N- difucohexaose I; and Lactodifucotetraose); non-fucosylated, non-sialylated oligosaccharides (z.e., Lacto-N-neotetraose); sialyl fucosyl oligosaccharides (z.e., 3 '-Sialyl-3 -fucosyllactose; Disialomonofucosyllacto-N-neohexaose; Monofucosylmonosialyllacto-N-octaose (sialyl Lea); Sialyllacto-N-fucohexaose II; Disialyllacto-N-fucopentaose II; Monofucosyldisialyllacto-N- tetraose); and sialyl oligosaccharides (z.e., 3'-Sialyllactose; 3-Sialyllactosamine; 6'-Sialyllactose; 6'-Sialyllactosamine; Sialyllacto-N-neotetraose c; Monosialyllacto-N-hexaose; Disialyllacto-N- hexaose I; Monosialyllacto-N-neohexaose I; Monosialyllacto-N-neohexaose II; Disialyllacto-N- neohexaose; Disialyllacto-N-tetraose; Disialyllacto-N-hexaose II; Sialyllacto-N-tetraose a; Disialyllacto-N-hexaose I; and Sialyllacto-N-tetraose b). Also useful are variants in which the glucose (Glc at the reducing end is replaced by N-acetylglucosamine (e.g., 2'-fucosyl-N- acetylglucosamine (2'-FLNac) is such a variant to 2'-fucosyllactose). These HMOs are described more fully in U.S. Patent Application No. 2009/0098240, which is herein incorporated by reference in its entirety. Other suitable examples of HMOs that may be included in the compositions according to the general inventive concepts include lacto-N-fucopentaose V, lacto- N-hexaose, para-lacto-N-hexaose, lacto-N-neohexaose, para-lacto-N-neohexaose, monofucosyllacto-N-hexaose II, isomeric fucosylated lacto-N-hexaose (1), isomeric fucosylated lacto-N-hexaose (3), isomeric fucosylated lacto-N-hexaose (2), difucosyl-para-lacto-N- neohexaose, difucosyl-para-lacto-N-hexaose, difucosyllacto-N-hexaose, lacto-N-neoocataose, para-lacto-N-octanose, iso-lacto-N-octaose, lacto-N-octaose, monofucosyllacto-neoocataose, monofucosyllacto-N-ocataose, difucosyllacto-N-octaose I, difucosyllacto-N-octaose II, difucosyllacto-N-neoocataose II, difucosyllacto-N-neoocataose I, lacto-N-decaose, trifucosyllacto-N-neooctaose, trifucosyllacto-N-octaose, trifucosyl-iso-lacto-N-octaose, lacto-N- difuco-hexaose II, sialyl-lacto-N-tetraose a, sialyl-lacto-N-tetraose b, sialyl-lacto-N-tetraose c, sialyl-fucosyl-lacto-N-tetraose I, sialyl-fucosyl-lacto-N-tetraose II, and disialyl-lacto-N-tetraose, and combinations thereof.

[0047] The HMOs are present in the nutritional compositions in total amounts of HMO in the composition (mg of HMO per mL of composition) of at least about 0.001 mg/mL to about 20 mg/mL, including about 0.001 mg/mL to about 20 mg/mL, 1 mg/mL to about 10 mg/mL, 2 mg/mL to about 10 mg/mL, and including from about 2.5 mg/mL to about 8.5 mg/mL. Typically, the amount of HMO in the nutritional composition will depend on the specific HMO or HMOs present and the amounts of other components in the nutritional compositions.

[0048] In certain exemplary aspects of the present disclosure, the nutritional composition includes at least one of 2’-FL, 3-FL, 3’-SL, 6'-SL, and LNT in a total amount of HMO of from about 0.001 mg/mL to about 20 mg/mL, including from about 0.01 mg/mL to about 20 mg/mL, including from about 0.1 mg/mL to about 20 mg/mL, about 0.5 mg/mL to about 20 mg/mL, about 1 mg/mL to about 20 mg/mL, about 2 mg/mL to about 20 mg/mL. about 3 mg/mL to about 20 mg/mL, and about 5 mg/mL to about 20 mg/mL. In certain exemplary aspects of the present disclosure, the nutritional composition includes at least one of 2’-FL, 3-FL, 3’-SL, 6'-SL, and LNT in a total amount of HMO of from about 0.1 g/L to about 20 mg/mL, including from about 0.1 mg/mL to about 15 mg/mL, including from about 0.1 mg/mL to about 10 mg/mL, including from about 0.1 mg/mL to about 9 mg/mL, including from about 0.1 mg/mL to about 8 mg/mL, including from about 0.1 mg/mL to about 7 mg/mL, including from about 0.1 mg/mL to about 6 mg/mL, including from about 0.1 mg/mL to about 5 mg/mL, including from about 0.1 mg/mL to about 4 mg/mL, including from about 0.1 mg/mL to about 3 mg/mL, including from about 0.1 mg/mL to about 2 mg/mL, including from about 0.5 mg/mL to about 4 mg/mL, including from about 0.5 mg/mL to about 3 mg/mL, including from about 1 mg/mL to about 2.5 mg/mL; and including from about 3.1 mg/mL to about 8.2 mg/mL.

[0049] In certain exemplary aspects of the present disclosure, the nutritional composition comprises an HMO component comprising at least one of a neutral HMO and an acidic HMO, including aspects that comprise both a neutral HMO and an acidic HMO. The neutral HMO may be a fucosylated or n-acetylated HMO, including but not limited to 2’ -FL and LNT.

[0050] In certain exemplary aspects, the HMO component comprises at least one neutral HMO and no acidic HMO. The neutral HMO may be a fucosylated or n-acetylated HMO, including but not limited to 2’-FL and LNT. In certain exemplary aspects, the neutral HMO is 2’- FL. In certain exemplary aspects, the neutral HMO is LNT.

[0051] In certain exemplary aspects, the HMO component comprises at least one acidic HMO and no neutral HMO. In certain exemplary aspects, the acidic HMO is a sialylated HMO, including but not limited to 6’-SL and/or 3’-SL.

[0052] In certain exemplary aspects, the nutritional composition comprises at least one of a fucosylated HMO, a N-acetylated HMO, and/or a sialylated HMO. In certain exemplary aspects of the present disclosure, the nutritional composition comprises a combination of a fucosylated HMO, a sialylated HMO, and an N-acetylated HMO. Specific non-limiting examples of suitable HMOs include 2’-FL, 6’-SL, and LNT.

[0053] In certain exemplary aspects of the present disclosure, the nutritional composition comprises a combination of 2’-FL, 3-FL, LNT, 3’-SL, and 6’-SL. In certain exemplary aspects of the present disclosure, the nutritional composition comprises 2’-FL in an amount up to 4.15 mg/mL, LNT in an amount up to 2.11 mg/mL, 3-FL in an amount up to 1.17 mg/mL, 3’-SL in an amount up to 0.36 mg/mL, and 6’-SL in an amount up to 0.44 mg/mL. In certain exemplary aspects, the combination of 2’-FL, 3-FL, LNT, 3’-SL, and 6’-SL is present in a weight ratio of approximately (50-55) : (10-15) : (20-30) : (2-7) : (2-7).

[0054] Nutritional compositions containing these combinations, as selected and defined herein, can be used to modulate or treat the conditions or diseases described herein, including but not limited to: boosting or supplementing an immune response in an individual; treating an infection in an individual; preventing an infection in an individual; and reducing or ameliorating symptoms of an infection in an individual. In certain aspects of the present disclosure, the nutritional composition includes at least one HMO in combination with a bovine immunoglobulin-containing protein component and one or more additional components as described herein such that the composition provides a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the conditions or diseases described herein.

Immunoglobulin(s)

[0055] The general inventive concepts relate to the use of a bovine immunoglobulin (Ig) protein component in combination with a human milk oligosaccharide component to prevent and/or treat infections, including viral infections. More specifically, the general inventive concepts relate to nutritional compositions which can be fed to infants, toddlers, children, adults, and the elderly and thereby prevent and/or treat infection by reducing the host cell infection and transmission of virus particles. Such administration is expected to achieve at least one of the following: prom ote/supplem ent the immune response in an individual; treat an infection in an individual; prevent an infection in an individual; and reduce or ameliorate symptoms of an infection in an individual. The general inventive concepts are based, at least in part on the discovery that Ig-enriched bovine milk protein fractions (e.g., the bovine immunoglobulin- containing protein component) in combination with HMOs greatly reduce, treat, or prevent respiratory virus infections. Such a benefit is not seen from common consumption of bovine milk products as typical consumption of bovine milk products does not generally achieve the amounts required to deliver the instant results as the concentrations of bovine Ig found in pasteurized cow milk are much lower than that of the general inventive concepts. Furthermore, the HMOs in the present invention are either absent from or at substantially lower concentrations in bovine milk than the current amounts according to the general inventive concepts.

[0056] Generally, IgA provides the major source of antibody protection conferred by human milk, however it is present at much lower concentrations in bovine milk. The general inventive concepts are based on the discovery of an unexpected synergy achieved through administration of a combination of a bovine immunoglobulin-containing protein component having an Ig-enriched bovine milk protein fraction and human milk oligosaccharides. This is distinguished from Ig isolated or otherwise obtained from serum. The nutritional compositions according to the general inventive concepts include a bovine immunoglobulin-enriched component comprising at least one bovine immunoglobulin, including, in certain aspects of the present disclosure, a combination including more than one bovine immunoglobulin (e.g., IgA, IgG, IgM).

[0057] The bovine immunoglobulin-containing protein component may be included in the nutritional compositions with HMOs alone, or in some aspects, in combination with other components to aid in ameliorating symptoms of the particular condition or disease (LCPUFAs, antioxidants, nucleotides, etc. as described herein. The bovine immunoglobulins are isolated, concentrated, or otherwise enriched from milk(s) sources secreted by bovids. In certain aspects, the bovine immunoglobulin-containing protein component has been enriched by a process (e.g., multiple filtration steps, including ultra-filtration, heating, and volume concentration) which achieves a higher concentration of immunoglobulins in the protein component than is seen in conventional milk protein components. In this way, in certain aspects, the nutritional composition may achieve the levels of immunoglobulin without additional direct supplementation of immunoglobulins. Bovine milk has been shown to have a relatively low ratio of IgATgG (e.g., < 0.2). In certain aspects, the bovine immunoglobulin-containing protein component according to the invention has been subjected to a process that enriches the ratio of IgATgG, increases the concentration of Ig relative to native materials, and/or a process that enriches the ratio of IgG to P-lactoglobulin. In certain aspects, the bovine immunoglobulin- containing protein component comprises an enriched ratio of IgA, including from about 10% to about 50%, including about 15% to about 45%, including certain aspects wherein a majority of the bovine immunoglobulin is IgA in the bovine immunoglobulin-containing protein component.

[0058] In certain exemplary aspects, the process includes at least one of ultrafiltration and heat treatment of the component. The bovine immunoglobulin-containing protein component according to the invention is more complex as it includes more native protein components than isolated or purified IgG/IgM ingredients and has not been merely supplemented to increase the amount of immunoglobulin or IgG. [0059] In certain aspects, a milk fraction or protein containing component which has been processed to enrich the IgG concentration may be employed in the methods and compositions described herein. Various enrichment techniques may be used. In certain aspects, the inventive concepts contemplate a method comprising (a) subjecting a milk fraction comprising skim milk and/or whey to microfiltration to defat and decaseinate the milk fraction and provide a sterile milk fraction permeate, (b) subjecting the sterile milk fraction permeate to ultrafiltration using a 700-800 kDa membrane and with a volume concentration factor (VCF) of at least about 5 to produce IgG-containing retenate, and (c) subjecting the IgG-containing retenate to additional ultrafiltration with a membrane sufficiently sized to remove lactose and mineral content.

[0060] In a more particular aspect, a milk fraction such as skim milk and/or whey (commonly referred to as cheese whey) is defatted and decaseinated via one or more microfiltration steps. In certain aspects, at least one microfiltration step employs at least one membrane of 2 pm or smaller in order to provide a sterile product and in certain aspects employs a membrane of 2 pm or smaller in conjunction with a membrane of 0.2 pm or smaller. In a more particular aspect, a first microfiltration step is conducted using a 1.4 pm microfiltration membrane and a subsequent microfiltration step is conducted using a 0.14 pm microfiltration membrane to remove fat and casein as well as bacteria and produce a sterile fraction. The microfiltration step(s) may be conducted at an elevated temperature, for example at a temperature of from about 30°C to about 60°C, including about 30°C to about 50°C, and/or with volume concentration, for example, with a volume concentration factor (VCF) of at least about 5, for example with a VCF of at least about 10, about 15, about 20, or about 25, including about 10 to about 20. In a specific aspect, a first microfiltration step is conducted using a 1.4 pm microfiltration membrane and a subsequent microfiltration step is conducted using a 0.14 pm microfiltration membrane with a VCF of at least about 10, at a temperature of from about 30°C to about 60°C, including about 30°C to about 50°C.

[0061] The permeate stream from the microfiltration step(s) is then subjected to first ultrafiltration using a 700-800 kDa membrane. In certain particular aspects, polymeric spiralwound membranes of 700 or 800 kDa are employed. The ultrafiltration may be conducted at a reduced temperature, for example at a temperature of from about 5 °C to about 20°C, including about 10 °C, and/or with volume concentration, for example, with a VCF of at least about 5, for example with a VCF of at least about 10, about 15, about 20, or about 5 to about 25, and including about 10 to about 20, to produce skim milk- and/or whey-derived fractions enriched for Ig (including for specific Ig’s such as IgG). In certain more specific aspects, the ultrafiltration step is conducted with a 700 or 800 kDa membrane and a VCF of at least about 10, including about 10 to about 20, at a temperature of from about 5°C to about 20°C, including about 10 °C. This ultrafiltration step removes a significant portion of P-lactoglobulin in the permeate and provides a desirable high IgG content and low P-lactoglobulin content in the retentate (i.e., a desirable ratio of IgG: P-lactoglobulin). In certain aspects, the weight ratio of IgG in the retenate from this ultrafiltration step to IgG in the permeate from the ultrafiltration step is greater than about 1.3, or greater than about 1.5, and the weight ratio of P-lactoglobulin in the retenate from the ultrafiltration step to P-lactoglobulin in the permeate from the ultrafiltration step is less than about 0.4, or less than about 0.3. The retentate may be subsequently processed through another (second) ultrafiltration step to remove lactose and mineral content and to increase protein content, for example using a 10 kDa membrane to provide a bovine immunoglobulin-containing protein component). The second ultrafiltration may also be conducted at a reduced temperature of from about 5°C to about 20°C, including about 10 °C. Depending on a desired format for use, the resulting bovine immunoglobulin-containing protein component liquid fractions may be subjected to a pasteurization heat treatment and/or spray-dried to produce an IgG-enriched powder.

[0062] The nutritional compositions as described herein will typically comprise total concentrations of bovine immunoglobulin proteins of from about 0.01 mg/mL to about 100 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.01 mg/mL to about 27 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.1 mg/mL to about 25 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.2 mg/mL to about 20 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.3 mg/mL to about 15 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.4 mg/mL to about 10 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 9 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 8 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 7 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 6 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 5 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 4 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 3 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 2.5 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 2 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 1.5 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.5 mg/mL to about 1 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.6 mg/mL to about 1 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.7 mg/mL to about 1 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.8 mg/mL to about 1 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 0.9 mg/mL to about 1 mg/mL. In certain exemplary aspects of the present disclosure, the bovine immunoglobulin is present in an amount of from about 1 mg/mL to about 1.5 mg/mL. In certain aspects, the bovine immunoglobulin comprises greater than 10% IgA, including 15% to about 50%, including concentrations of greater than 0.4 mg/mL.

[0063] It is important to distinguish between methods of bovine antibody (bAb) detection for purposes of the specified concentration ranges. Methods which detect protein fragments (peptides) or intact proteins which have been denatured by typical protein enrichment and processing techniques may not be able to distinguish between inherent bAbs in processed milk products and the bioactive versions that make up the general inventive concepts. A typical example of a method of this type is mass spectrometry. Appropriate analytical methods for determining bAb concentrations rely on recognition of intact protein structures. One example is the ELISA test.

[0064] In certain exemplary aspects of the present disclosure, the human milk oligosaccharide and bovine immunoglobulin are present in the nutritional composition in a particular weight ratio including from 1 :99 to 99: 1. In certain exemplary aspects of the present disclosure, the human milk oligosaccharide and bovine immunoglobulin are present in the nutritional composition in a weight ratio of 1 :20 to 20: 1. In certain exemplary aspects of the present disclosure, the human milk oligosaccharide and bovine immunoglobulin are present in the nutritional composition in a weight ratio of 1 : 10 to 10: 1. In certain exemplary aspects of the present disclosure, the human milk oligosaccharide and bovine immunoglobulin are present in the nutritional composition in a weight ratio of 1 : 5 to 5 : 1.

Long Chain Polyunsaturated Fatty Acids

[0065] In addition to the HMO and bovine immunoglobulin-containing protein component combination described above, the nutritional compositions according to the general inventive concepts may include Long Chain Polyunsaturated Fatty Acids (LCPUFAs). LCPUFAs are included in the nutritional compositions to provide nutritional support, as well as to modulate or treat the conditions or diseases described herein, including modulating one or more aspects of viral infection. In certain aspects of the present disclosure, the nutritional composition includes a combination of the HMOs and bovine immunoglobulin-containing protein component and one or more LCPUFAs such that the composition provides a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the conditions or diseases described herein.

[0066] Exemplary LCPUFAs for use in the nutritional compositions include, for example, co-3 LCPUFAs and co-6 LCPUFAs. Specific LCPUFAs include docosahexaenoic acid (DHA), eicosapentaenoic acid (EP A), docosapentaenoic acid (DPA), arachidonic acid (ARA), linoleic acid, linolenic acid (alpha linolenic acid) and gamma-linolenic acid derived from oil sources such as plant oils, marine plankton, fungal oils, krill oil and fish oils. In one particular aspect, the LCPUFAs are derived from fish oils such as menhaden, salmon, anchovy, cod, halibut, tuna, or herring oil. Particularly preferred LCPUFAs for use in the nutritional compositions include DHA, ARA, EP A, DPA, and combinations thereof.

[0067] In order to reduce potential side effects of high dosages of LCPUFAs including DHA, ARA, EP A, DPA, in the nutritional compositions, the content of DHA, ARA, EP A, DPA, preferably does not exceed 5% by weight of the total fat content, including below 2% by weight of the total fat content, and including below 1% by weight of the total fat content in the nutritional composition.

[0068] The LCPUFA may be provided as free fatty acids, in triglyceride form, in diglyceride form, in monoglyceride form, in phospholipid form, in esterified form or as a mixture of one or more of the above, preferably in triglyceride form.

[0069] The nutritional compositions as described herein will typically comprise total concentrations of ARA, DHA, EP A, and DPA of DHA, ARA, EP A, DPA, of from about 0.001 g/L to about 1 g/L or more, including from about 0.01 g/L to about 1 g/L, and about 0.1 g/L to about 1 g/L.

[0070] In an exemplary aspect, the nutritional compositions include a long chain polyunsaturated fatty acid component comprising DHA and ARA in a concentration of from about 0.17 mg/mL to about 0.33 mg/mL, including from about 0.17 mg/mL to about 0.26 mg/mL of ARA and DHA. In an exemplary aspect, the nutritional compositions include DHA in a concentration of from about 0.025 mg/mL to about 0.130 mg/mL. In another aspect, the nutritional compositions include ARA in a concentration of from about 0.080 mg/mL to about 0.350 mg/mL. In yet another aspect, the nutritional compositions include combinations of DHA and ARA such that the ratio of DHA to ARA ranges from about 1 :4 to about 1 :2.

Antioxidants

[0071] Additionally, the nutritional compositions may comprise one or more antioxidants in combination with the HMOs and bovine immunoglobulin-containing protein component to provide nutritional support, as well as to provide a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the conditions or diseases described herein. In some aspects of the present disclosure, the HMO and bovine immunoglobulin-containing protein component blend is used in combination with carotenoids (and specifically lutein, beta-carotene, zeaxanthin and/or lycopene) to provide the synergistic effect.

[0072] Any antioxidants suitable for oral administration may be included for use in the nutritional compositions according to the general inventive concepts, including, for example, vitamin A, vitamin E, vitamin C, retinol, tocopherol, and carotenoids, including lutein, betacarotene, zeaxanthin, and lycopene, and combinations thereof, for example.

[0073] As noted, the antioxidants for use in the nutritional compositions may be used with the HMO and bovine immunoglobulin-containing protein component alone or in combination with LCPUFAs, and/or nucleotides. In certain aspects of the present disclosure, the nutritional composition includes a combination of the HMOs and bovine immunoglobulin- containing protein component and one or more of LCPUFAs, antioxidants, and nucleotides as described herein such that the composition provides a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the aforementioned conditions or diseases.

[0074] It is generally preferable that the nutritional compositions comprise at least one carotenoid selected from lutein, lycopene, zeaxanthin, and beta-carotene to provide a total amount of carotenoid of from about 0.001 pg/mL to about 10 pg/mL. More particularly, the nutritional compositions comprise lutein in an amount of from about 0.001 pg/mL to about 10 pg/mL, including from about 0.001 pg/mL to about 5 pg/mL, including from about 0.001 pg/mL to about 1 pg/mL, including from about 0.001 pg/mL to about 0.5 pg/mL, including from about 0.001 pg/mL to about 0.190 pg/mL, including from about 0.001 pg/mL to about 0.140 pg/mL, and also including from about 0.044 pg/mL to about 5 pg/mL of lutein. It is also generally preferable that the nutritional compositions comprise from about 0.001 pg/mL to about 10 pg/mL, including from about 0.001 pg/mL to about 5 pg/mL, from about 0.001 pg/mL to about 0.0130 pg/mL, including from about 0.001 pg/mL to about 0.0075 pg/mL, and also including from about 0.0185 pg/mL to about 5 pg/mL of lycopene. It is also generally preferable that the nutritional compositions comprise from about 0.001 pg/mL to about 10 pg/mL, including from about 0.001 pg/mL to about 5 pg/mL, including from about 0.001 pg/mL to about 1 pg/mL, including from about 0.001 pg/mL to about 0.1 pg/mL, and also including from about 0.034 pg/mL to about 5 pg/mL of beta-carotene. It should be understood that any combination of these amounts of beta-carotene, lutein, zeaxanthin, and lycopene can be included in the nutritional compositions according to the general inventive concepts. In certain aspects, the nutritional composition may be free of any one or more of beta-carotene, lutein, zeaxanthin, and lycopene. The composition may further be substantially free of other carotenoids. Other carotenoids may optionally be included in the nutritional compositions as described herein. Any one or all of the carotenoids included in the nutritional compositions described herein may be from a natural source or artificially synthesized.

[0075] Each of the carotenoids in the selected combinations can be obtained from any known or otherwise suitable material source for use in nutritional compositions, and each can be provided individually, or all together, or in any combination and from any number of sources, including sources such as multivitamin premixes containing other vitamins or minerals in combination with one or more of the carotenoids as described herein. Non-limiting examples of some suitable sources of lutein, lycopene, beta-carotene, or combinations thereof include LycoVit® lycopene (available from BASF, Mount Olive, N. J.), Lyc-O-Mato® tomato extract in oil, powder, or bead form (available from LycoRed Corp., Orange, N.J.), beta-carotene, lutein, or lycopene (available from DSM Nutritional Products, Parsippany, N.J.), FloraGLO® lutein (available from Kemin Health, Des Moines, Iowa), Xangold® Natural Lutein Esters (available from Cognis, Cincinnati, Ohio), and Lucarotin® beta-carotene (available from BASF, Mount Olive, N.J.).

RRR- Alpha-Tocopherol

[0076] The nutritional compositions described herein may include RRR-alpha- tocopherol in combination with the HMOs and bovine immunoglobulin-containing protein component to provide nutritional support, as well as to provide a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the conditions or diseases described herein. [0077] In certain exemplary aspects of the present disclosure, the nutritional compositions contain RRR-alpha-tocopherol in concentrations of at least about 5 mg/L, including at least about 7 mg/L, including at least about 8 mg/L, including at least about 9 mg/L, including at least about 10 mg/L, including from at least about 5 mg/L to about 100 mg/L, including from at least about 7 mg/L to about 50 mg/L, including from about 7 mg/L to about 40 mg/L, and including from about 7 mg/L to about 20 mg/L. The total amounts of RRR-alpha- tocopherol include both exogenous and inherent sources of RRR-alpha-tocopherol, as noted below.

[0078] As used herein, the term “RRR-alpha-tocopherol” refers both to exogenous sources and inherent sources of RRR-alpha-tocopherol and RRR-alpha-tocopherol acetate that are present in a nutritional composition. Inherent sources include RRR-alpha-tocopherol that is inherently present in components that are present in a nutritional composition and may include for example, various oils and fats. Exogenous sources of RRR-alpha-tocopherol include RRR- alpha-tocopherol that is added to the nutritional composition not as part of another component.

[0079] Tocopherols, generically referred to as vitamin E, are available in four forms: alpha-, beta-, gamma-, and delta-, which differ in the number and position of the methyl groups on the chroman ring, as shown by the structure below and Table 1.

TABLE 1

Structure of Natural Tocopherols

Compound Ri R2 R3 alpha-tocopherol Me Me Me beta-tocopherol Me H Me gamma-tocopherol H Me Me delta-tocopherol H H Me

[0080] Tocopherols can exist in a number of stereoisomeric forms depending on the chirality of the phytyl tail. Of the alpha-tocopherols, RRR-alpha-tocopherol (also referred to as “natural vitamin E”) has the greatest biological activity and is reported to be the dominant form of alpha-tocopherol in the brain. RRR-alpha-tocopherol is a single stereoisomer whereas synthetic vitamin E (all-rac-alpha-tocopherol or tocopherol acetate) is an equimolar mixture of eight isomers, only one of which is RRR-alpha-tocopherol. The fact that the dominant form of alpha-tocopherol is RRR-alpha-tocopherol (based on animal studies) strongly suggests that the other seven chiral isomers are absorbed at a lower rate by the brain or oxidized at a faster rate.

Nucleotides

[0081] The nutritional compositions described herein may include nucleotides in combination with the HMOs and bovine immunoglobulin-containing protein component to provide nutritional support, as well as to provide a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the conditions or diseases described herein. In certain exemplary aspects of the present disclosure, the “nucleotides” comprise nucleotides and/or nucleotide precursors selected from nucleosides, purine bases, pyrimidine bases, ribose and deoxyribose. The nucleotide may be in monophosphate, diphosphate, or triphosphate form. The nucleotide may be a ribonucleotide or a deoxyribonucleotide. The nucleotides may be monomeric, dimeric, or polymeric (including RNA and DNA). The nucleotide may be present in the nutritional composition as a free acid or in the form of a salt, preferably a monosodium salt. In some aspects of the present disclosure, the nutritional composition includes a combination of HMOs, bovine immunoglobulin-containing protein component and nucleotides such that the composition provides a synergistic benefit to the end user, such as a synergistic benefit in modulating or treating one or more of the aforementioned conditions or diseases.

[0082] Suitable nucleotides and/or nucleosides for use in the nutritional compositions include one or more of cytidine 5 '-monophosphate, uridine 5 '-monophosphate, adenosine 5'- monophosphate, guanosine 5 '-1 -monophosphate, and/or inosine 5 '-monophosphate, more preferably cytidine 5 '-monophosphate, uridine 5 '-monophosphate, adenosine 5 '-monophosphate, guanosine 5 '-monophosphate, and inosine 5 '-monophosphate.

[0083] In an exemplary aspect, the nucleotides are present in the nutritional composition in a total amount of about 50 mg/L to about 300 mg/mL, including about 70 mg/L to about 300 mg/L of the nutritional composition. In an exemplary aspect, the nucleotides are present in the nutritional composition in a total amount of about 70 mg/L of the nutritional composition. In certain exemplary aspects of the present disclosure, the nucleotides comprise 40- 50% cytidine 5 ’monophosphate, about 15-20% uridine 5 '-monophosphate, about 12-18% adenosine 5 '-monophosphate and about 20-25% guanosine 5 '-monophosphate by total weight of nucleotides. In certain exemplary aspects of the present disclosure, the nucleotides comprise about 43% cytidine 5 '-monophosphate, about 18.5% uridine 5 '-monophosphate, about 16.5% adenosine 5 '-monophosphate and about 22% guanosine 5 '-monophosphate by total weight of nucleotides.

[0084] In an exemplary aspect, the nucleotides are present in the nutritional compositions in a total amount of about 50 mg/L to about 300 mg/mL of the nutritional composition and comprise about 25 to 39 mg/L of cytidine 5 '-monophosphate; 10 to 21 mg/L of uridine 5 '-monophosphate; 10 to 15 mg/L of adenosine 5 '-monophosphate; and 12 to 20 mg/L of guanosine 5 '-monophosphate.

[0085] In an exemplary aspect, the nucleotides are present in the weight ratio of cytidine 5 '-monophosphate: uridine 5 '-monophosphate is from about 1.5: 1 to about 2.6: 1; of cytidine 5 '-monophosphate: adenosine 5 '-monophosphate is from about 2: 1 to about 3.9: 1; and of cytidine 5 '-monophosphate: guanosine 5 '-monophosphate is from about 1.75: 1 to about 2.8:1.

[0086] In addition to the specific functional ingredients described herein, the nutritional composition includes one or more ingredients that help satisfy the individual’s nutritional requirements. The optional nutrients can provide up to about 1000 kcal of energy per serving or dose, including from about 25 to about 900 kcal, from about 75 to about 700 kcal, from about 150 to about 500 kcal, from about 350 to about 500 kcal, or from about 200 to about 300 kcal. [0087] The nutritional compositions may be formulated with sufficient kinds and amounts of nutrients to provide a sole, primary, or supplemental source of nutrition, or to provide a specialized nutritional composition for use in individuals afflicted with specific diseases or conditions or with a targeted nutritional benefit as described below.

[0088] The nutritional compositions including the HMO or HMOs may be formulated to include at least one of protein, fat, and carbohydrate. In many aspects, the nutritional compositions will include the HMO or HMOs with protein, carbohydrate and fat. Although total concentrations or amounts of the fat, protein, and carbohydrates may vary depending upon the product type (i.e., nutritional formula), product form (/.< ., nutritional solid, powder, ready-to- feed liquid, or concentrated liquid) and targeted dietary needs of the intended user, such concentrations or amounts most typically fall within one of the following embodied ranges, inclusive of any other essential fat, protein, and/or carbohydrate ingredients as described herein.

Macronutrients

[0089] In certain aspects, the nutritional composition will include at least one of protein, fat, and carbohydrate. In many aspects, the nutritional composition will include protein, fat, and carbohydrate.

[0090] Where present, carbohydrate concentrations most typically will range from about 5% to about 40%, including from about 7% to about 30%, including from about 10% to about 25%, by weight of the nutritional composition. Where present, fat concentrations most typically range from about 1% to about 30%, including from about 2% to about 15%, and also including from about 3% to about 10%, by weight of the nutritional composition. Where present, protein concentrations most typically range from about 0.5% to about 30%, including from about 1% to about 15%, and also including from about 2% to about 10%, by weight of the nutritional composition.

[0091] The amount of any or all of the carbohydrates, fats, and proteins in any of the nutritional compositions (e.g., infant formula) described herein may also be characterized as a percentage of total calories in the nutritional composition as set forth in the following table. These macronutrients for nutritional compositions according to the general inventive concepts are most typically formulated within any of the caloric ranges (embodiments A-F) described in the following table (each numerical value is preceded by the term “about”).

Table 2. Exemplary macronutrient profiles of nutritional compositions

Fat

[0092] The nutritional compositions according to the general inventive concepts may, in addition to the LCPUFA’s, comprise an additional source or sources of fat. Suitable additional sources of fat for use herein include any fat or fat source that is suitable for use in an oral nutritional composition and is compatible with the essential elements and features of such compositions. Most typically the fat may be an emulsified fat, concentrations of which may range from about 1% to about 30%, including from about 2% to about 15%, and also including from about 4% to about 10%, by weight. In an exemplary aspect of the present disclosure, the additional fat is derived from short chain fatty acids. [0093] Additional non-limiting examples of suitable fats or sources thereof for use in the nutritional compositions described herein include coconut oil, fractionated coconut oil, soybean oil, com oil, olive oil, safflower oil, high oleic safflower oil, oleic acids (EMERSOL 6313 OLEIC ACID, Cognis Oleochemicals, Malaysia), MCT oil (medium chain triglycerides), sunflower oil, high oleic sunflower oil, palm and palm kernel oils, palm olein, canola oil, marine oils, fish oils, fungal oils, algae oils, cottonseed oils, and combinations thereof. Lipid sources of arachidonic acid and docosahexaenoic acid include, but are not limited to, marine oil, egg yolk oil, and fungal or algal oil.

[0094] Numerous commercial sources for these fats are readily available and known to one practicing the art. For example, soy and canola oils are available from Archer Daniels Midland of Decatur, Ill. Corn, coconut, palm and palm kernel oils are available from Premier Edible Oils Corporation of Portland, Organ. Fractionated coconut oil is available from Henkel Corporation of LaGrange, Ill. High oleic safflower and high oleic sunflower oils are available from SVO Specialty Products of Eastlake, Ohio. Marine oil is available from Mochida International of Tokyo, Japan. Olive oil is available from Anglia Oils of North Humberside, United Kingdom. Sunflower and cottonseed oils are available from Cargil of Minneapolis, Minn. Safflower oil is available from California Oils Corporation of Richmond, Calif.

[0095] In addition to these food grade oils, structured lipids may be incorporated into the food product if desired. Structured lipids are known in the art. A concise description of structured lipids can be found in INFORM, Vol. 8, No. 10, page 1004; entitled Structured lipids allow fat tailoring (October 1997). Also see U.S. Pat. No. 4,871,768. Structured lipids are predominantly triacylglycerols containing mixtures of medium and long chain fatty acids on the same glycerol nucleus. Structured lipids and their use in enteral formula are also described in U.S. Pat. Nos. 6,194,379 and 6,160,007.

[0096] Optionally, co-3 fatty acids may comprise up to approximately 5% of the oil blend, preferably the co-3 fatty acids largely consist of the longer chain forms, eicosapentaenoic acid (EP A) and docosahexaenoic acid (DHA). Dietary oils used in the preparation of the nutritional composition generally contain co-3 fatty acids in the triglyceride form and include, but are not limited to canola, medium chain triglycerides, fish, soybean, soy lecithin, corn, safflower, sunflower, high-oleic sunflower, high-oleic safflower, olive, borage, black currant, evening primrose and flaxseed oil. Optionally, the weight ratio of co-6 fatty acids to co-3 fatty acids in the lipid blend according to the invention is about 0.1 to 3.0. The daily delivery of co-3 fatty acids should be at least 450 mg and may vary depending on body weight, sex, age and medical condition of the individual. As mentioned, higher levels are desired for adult human consumption: for example, from about 0.5 to 50 gm daily, more preferably from about 2.5 to 5 gm daily

Protein

[0097] The nutritional compositions according to the general inventive concepts include protein, both as part of the bovine immunoglobulin-containing protein component as well as additional optional protein and protein sources. In addition to the bovine immunoglobulin-containing protein component, any protein source that is suitable for use in oral nutritional composition and is compatible with the essential elements and features of such formulas is suitable for use in the nutritional compositions.

[0098] Non-limiting examples of suitable proteins or sources thereof for use in the nutritional compositions include hydrolyzed, partially hydrolyzed or non-hydrolyzed proteins or protein sources, which may be derived from any known or otherwise suitable source such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, com), vegetable (e.g., soy) or combinations thereof. Non-limiting examples of such proteins include milk protein isolates, milk protein concentrates as described herein, casein protein isolates, extensively hydrolyzed casein, whey protein, sodium or calcium caseinates, whole cow milk, partially or completely defatted milk, soy protein isolates, and soy protein concentrates. In an aspect, the nutritional compositions include a protein source derived from milk proteins of human and/or bovine origin.

[0099] Those of ordinary skill in the art will recognize that total protein content will vary depending on the target consumer and depending on nutritional needs. In certain exemplary aspects of the present disclosure, an infant formula is contemplated having up to about 21 grams of protein per liter. In certain exemplary aspects of the present disclosure, an adult nutritional composition is contemplated having up to about 90 grams of protein per liter. [00100] In an exemplary aspect of the present disclosure, the protein source is a hydrolyzed protein, /.< ., a protein hydrolysate. In this context, the terms “hydrolyzed protein” or “protein hydrolysates” are used interchangeably herein and include extensively hydrolyzed proteins, wherein the degree of hydrolysis is most often at least about 20%, including from about 20% to about 80%, and also including from about 30% to about 80%, even more preferably from about 40% to about 60%. The degree of hydrolysis is the extent to which peptide bonds are broken by a hydrolysis method. The degree of protein hydrolysis for purposes of characterizing the extensively hydrolyzed protein component of these embodiments is easily determined by one of ordinary skill in the formulation arts by quantifying the amino nitrogen to total nitrogen ratio (AN/TN) of the protein component of the selected liquid formulation. The amino nitrogen component is quantified by USP titration methods for determining amino nitrogen content, while the total nitrogen component is determined by the Tecator Kjeldahl method, all of which are well known methods to one of ordinary skill in the analytical chemistry art.

[00101] Suitable hydrolyzed proteins include soy protein hydrolysate, casein protein hydrolysate, whey protein hydrolysate, rice protein hydrolysate, potato protein hydrolysate, fish protein hydrolysate, egg albumen hydrolysate, gelatin protein hydrolysate, combinations of animal and vegetable protein hydrolysates, and combinations thereof. Particularly preferred protein hydrolysates include whey protein hydrolysate and hydrolyzed sodium caseinate.

[00102] When used in the nutritional compositions, the protein source may include at least about 1% (by weight total protein) protein hydrolysate, including from about 2% to 100% (by weight total protein) protein hydrolysate, including from about 2% to 10% (by weight total protein) protein hydrolysate, including from about 5% to 100% (by weight total protein) protein hydrolysate, including from about 10% to 100% (by weight total protein) protein hydrolysate, and including from about 20% to about 80% (by weight total protein) protein hydrolysate, and including about 50% (by weight total protein) protein hydrolysate. In an aspect, the nutritional composition includes 100% (by weight total protein) protein hydrolysate. Carbohydrate

[00103] The nutritional compositions according to the general inventive concepts may, in addition to the HMOs, include carbohydrates that are suitable for use in an oral nutritional composition and are compatible with the essential elements and features of such compositions.

[00104] Non-limiting examples of suitable carbohydrates or sources thereof for use in the nutritional compositions described herein include maltodextrin, hydrolyzed or modified starch or cornstarch, glucose polymers, com syrup, corn syrup solids, rice-derived carbohydrates, pea-derived carbohydrates, potato-derived carbohydrates, tapioca, sucrose, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), artificial sweeteners e.g., sucralose, acesulfame potassium, stevia) and combinations thereof. A particularly desirable carbohydrate is a low dextrose equivalent (DE) maltodextrin.

Optional Ingredients

[00105] The nutritional compositions according to the general inventive concepts may further comprise other optional components that may modify the physical, chemical, aesthetic or processing characteristics of the products or serve as pharmaceutical or additional nutritional components when used in the targeted population. Many such optional ingredients are known or otherwise suitable for use in medical food or other nutritional compositions or pharmaceutical dosage forms and may also be used in the compositions herein, provided that such optional ingredients are safe for oral administration and are compatible with the essential and other ingredients in the selected product form.

[00106] Non-limiting examples of such optional ingredients include preservatives, emulsifying agents, buffers, nucleotides, fructooligosaccharides, fiber, galactooligosaccharides, polydextrose, and other prebiotics, probiotics, pharmaceutical actives, anti-inflammatory agents, additional nutrients as described herein, colorants, flavors, thickening agents and stabilizers, emulsifying agents, lubricants, and so forth.

[00107] The nutritional compositions may further comprise a sweetening agent, preferably including at least one sugar alcohol such as maltitol, erythritol, sorbitol, xylitol, mannitol, isomalt, and lactitol, and also preferably including at least one artificial or high potency sweetener such as acesulfame K, aspartame, sucralose, saccharin, stevia, and tagatose. These sweetening agents, especially as a combination of a sugar alcohol and an artificial sweetener, are especially useful in formulating liquid beverage embodiments according to the general inventive concepts having a desirable favor profile. These sweetener combinations are especially effective in masking undesirable flavors sometimes associated with the addition of vegetable proteins to a liquid beverage. Optional sugar alcohol concentrations in the nutritional compositions may range from at least 0.01%, including from about 0.1% to about 10%, and also including from about 1% to about 6%, by weight of the nutritional compositions. Optional artificial sweetener concentrations may range from about 0.01%, including from about 0.05% to about 5%, also including from about 0.1% to about 1.0%, by weight of the nutritional compositions.

[00108] A flowing agent or anti-caking agent may be included in the nutritional compositions as described herein to retard clumping or caking of the powder over time and to make a powder composition flow easily from its container. Any known flowing or anti-caking agents that are known or otherwise suitable for use in a nutritional powder or product form are suitable for use herein, non-limiting examples of which include tricalcium phosphate, silicates, and combinations thereof. The concentration of the flowing agent or anti-caking agent in the nutritional composition varies depending upon the product form, the other selected ingredients, the desired flow properties, and so forth, but most typically range from about 0.1% to about 4%, including from about 0.5% to about 2%, by weight of the nutritional composition.

[00109] A stabilizer may also be included in the nutritional compositions. Any stabilizer that is known or otherwise suitable for use in a nutritional composition is also suitable for use herein, some non-limiting examples of which include gums such as xanthan gum. The stabilizer may represent from about 0.1% to about 5.0%, including from about 0.5% to about 3%, including from about 0.7% to about 1.5%, by weight of the nutritional composition.

[00110] The nutritional compositions may further comprise any of a variety of other vitamins or related nutrients, non-limiting examples of which include vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12, niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, salts and derivatives thereof, and combinations thereof. The food products preferably include, but are not limited to, the following vitamins and minerals: calcium, phosphorus, sodium, chloride, magnesium, manganese, iron, copper, zinc, selenium, iodine, chromium, molybdenum, conditionally essential nutrients m- inositol, carnitine and taurine, and Vitamins A, C, D, E, K and the B complex, and mixtures thereof.

[00111] The nutritional compositions may further comprise any of a variety of other additional minerals, non-limiting examples of which include calcium, phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, chloride, and combinations thereof.

[00112] The nutritional compositions also may contain fiber and stabilizers. Suitable sources of fiber and/or stabilizers include, but are not limited to, xanthan gum, guar gum, gum arabic, gum ghatti, gum karaya, gum tracacanth, agar, furcellaran, gellan gum, locust bean gum, pectin, low and high methoxy pectin, oat and barley glucans, carrageenans, psyllium, gelatin, microcrystalline cellulose, CMC (sodium carboxymethylcellulose), methylcellulose hydroxypropyl methyl cellulose, hydroxypropyl cellulose, DATEM (diacetyl tartaric acid esters of mono- and diglycerides), dextran, carrageenans, FOS (fructooligosaccharides), and mixtures thereof. Numerous commercial sources of soluble dietary fibers are available. For example, gum arabic, hydrolyzed carboxymethylcellulose, guar gum, pectin and the low and high methoxy pectins are available from TIC Gums, Inc. of Bel camp, Md. The oat and barley glucans are available from Mountain Lake Specialty Ingredients, Inc. of Omaha, Nebr. Psyllium is available from the Meer Corporation of North Bergen, N.J. while the carrageenan is available from FMC Corporation of Philadelphia, Pa.

[00113] In addition to fiber, the nutritional compositions may also contain oligosaccharides such as fructooligosaccharides (FOS) or galactooligosaccharides (GOS). Oligosaccharides are rapidly and extensively fermented to short chain fatty acids by anaerobic microorganisms that inhabit the large bowel. These oligosaccharides are preferential energy sources for most Bifidobacterium species, but are not utilized by potentially pathogenic organisms such as Clostridium perfmgens, C. difficile, or Eschericia coli. [00114] The nutritional compositions may additionally comprise one or more thickeners (i.e., thickening agents). The addition of thickeners reduces the incidences of paresthesia by inducing the feeling of satiety, which prolongs gastric transit time as discussed above.

Nutritional Composition Forms

[00115] The nutritional compositions according to the general inventive concepts may be formulated and administered in any known or otherwise suitable oral product form. Any solid, liquid, semi-solid, and semi-liquid, or powder product form, including combinations or variations thereof, are suitable for use herein, provided that such forms allow for safe and effective oral delivery to the individual of the essential ingredients as also defined herein.

[00116] The nutritional compositions according to the general inventive concepts include an HMO blend as described herein. The compositions may include an HMO blend with bovine immunoglobulin-containing protein component alone or in combination with other functional ingredients, such as LCPUFAs, nucleotides, and antioxidants, such as carotenoids, and vitamins, as discussed herein.

[00117] The nutritional compositions may be in any product form comprising the ingredients described herein, and which is safe and effective for oral administration. The nutritional compositions may be formulated to include only the ingredients described herein, or may be modified with optional ingredients to form a number of different product forms.

[00118] The nutritional compositions according to the general inventive concepts are desirably formulated as dietary product forms, which are defined herein as those embodiments comprising the ingredients according to the general inventive concepts in a product form that then contains at least one of fat, protein, and carbohydrate, and preferably also contains vitamins, minerals, or combinations thereof. The nutritional compositions will comprise at least HMOs and bovine immunoglobulin-containing protein component, desirably in combination with at least one of fat, vitamins, minerals, and additional protein to produce a nutritional composition.

Nutritional Liquids [00119] Liquid nutritional compositions include both concentrated and ready -to-feed nutritional liquids. These nutritional liquids are most typically formulated as suspensions or emulsions, although other liquid forms are within the scope of the general inventive concepts. Nutritional compositions in the form of emulsions suitable for use may be aqueous emulsions comprising proteins, fats, and carbohydrates. These emulsions are generally flowable or drinkable liquids at from about 1° C. to about 25° C. and are typically in the form of oil-in-water, water-in-oil, or complex aqueous emulsions, although such emulsions are most typically in the form of oil-in-water emulsions having a continuous aqueous phase and a discontinuous oil phase.

[00120] The nutritional liquids may be and typically are shelf stable. The nutritional emulsions typically contain up to about 95% by weight of water, including from about 50% to about 95%, also including from about 60% to about 90%, and also including from about 70% to about 85%, of water by weight of the nutritional emulsions. The nutritional emulsions may have a variety of product densities, but most typically have a density greater than about 1.03 g/mL, including greater than about 1.04 g/mL, including greater than about 1.055 g/mL, including from about 1.06 g/mL to about 1.12 g/mL, and also including from about 1.085 g/mL to about 1.10 g/mL.

[00121] The nutritional liquid compositions may have a caloric density tailored to the nutritional needs of the ultimate user, although in most instances the emulsions comprise generally at least 19 kcal/fl oz (660 kcal/liter), more typically from about 20 kcal/fl oz (675-680 kcal/liter) to about 25 kcal/fl oz (820 kcal/liter), even more typically from about 20 kcal/fl oz (675-680 kcal/liter) to about 24 kcal/fl oz (800-810 kcal/liter). Generally, the 22-24 kcal/fl oz formulas are more commonly used in preterm or low birth weight infants, and the 20-21 kcal/fl oz (675-680 to 700 kcal/liter) formulas are more often used in term infants. In some aspects of the present disclosure, the emulsion may have a caloric density of from about 50-100 kcal/liter to about 660 kcal/liter, including from about 150 kcal/liter to about 500 kcal/liter. In an exemplary aspect of the present disclosure, the emulsion may have a caloric density of 25, or 50, or 75, or 100 kcal/liter. [00122] The nutritional emulsion may have a pH ranging from about 2.5 to about 8, but are most advantageously in a range of from about 4.5 to about 7.5, including from about 5.5 to about 7.3, including from about 6.2 to about 7.2.

[00123] Although the serving size for the nutritional liquid can vary depending upon a number of variables, a typical serving size is generally at least about 1 mL, or even at least about 2 mL, or even at least about 5 mL, or even at least about 10 mL, or even at least about 25 mL, including ranges from about 1 mL to about 360 mL, including from about 30 mL to about 250 mL, and including from about 60 mL to about 240 mL.

Nutritional Solids

[00124] The nutritional compositions in the form of nutritional solids may be in any solid form but are typically in the form of flowable or substantially flowable particulate compositions, or at least particulate compositions. Particularly suitable nutritional solid product forms include spray dried, agglomerated and/or dry-blended powder compositions. The compositions can easily be scooped and measured with a spoon or similar other device, and can easily be reconstituted by the intended user with a suitable aqueous liquid, typically water, to form a nutritional composition for immediate oral or enteral use. In this context, “immediate” use generally means within about 48 hours, most typically within about 24 hours, preferably right after reconstitution.

[00125] The nutritional powders may be reconstituted with water prior to use to a caloric density tailored to the nutritional needs of the ultimate user, similar to that described above for liquid nutritional compositions.

Methods of Manufacture

[00126] The nutritional compositions according to the general inventive concepts may be prepared by any known or otherwise effective manufacturing technique for preparing the nutritional compositions. Many such techniques are known for any given product form such as nutritional liquids or powders and can easily be applied by one of ordinary skill in the art to the nutritional compositions described herein. [00127] The nutritional compositions (e.g., infant formulas) according to the general inventive concepts can therefore be prepared by any of a variety of known or otherwise effective formulation or manufacturing methods. In one suitable manufacturing process, for example, at least three separate slurries are prepared, including a protein-in-fat (PIF) slurry, a carbohydrate- mineral (CHO-MIN) slurry, and a protein-in-water (PIW) slurry. The PIF slurry is formed by heating and mixing the oil (e.g., canola oil, com oil, etc.) and then adding an emulsifier (e.g., lecithin), fat soluble vitamins, and a portion of the total protein (e.g., milk protein concentrate, immunoglobulin containing protein component, etc.) with continued heat and agitation. The CHO-MIN slurry is formed by adding with heated agitation to water: minerals (e.g., potassium citrate, dipotassium phosphate, sodium citrate, etc.), trace and ultra-trace minerals (TM/UTM premix), thickening or suspending agents (e.g., avicel, gellan, and carrageenan). The resulting CHO-MIN slurry is held for 10 minutes with continued heat and agitation before adding additional minerals (e.g., potassium chloride, magnesium carbonate, potassium iodide, etc.), and/or carbohydrates (e.g., HMOs, fructooligosaccharide, sucrose, com syrup, etc.). The PIW slurry is then formed by mixing with heat and agitation the remaining protein, if any.

[00128] The resulting slurries are then blended together with agitation and the pH adjusted to 6.6-7.0, after which the composition is subjected to a process such as vat pasteurization processing during which the composition is emulsified and homogenized. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range, if necessary, flavors are added, and water is added to achieve the desired total solid level. This emulsion can then be further diluted, heat-treated, and packaged to form a ready-to-feed or concentrated liquid, or it can be heat-treated and subsequently processed and packaged as a reconstitutable powder, e.g., spray dried, drymixed, agglomerated.

[00129] The nutritional solid, such as a spray dried nutritional powder or drymixed nutritional powder, may be prepared by any collection of known or otherwise effective techniques, suitable for making and formulating a nutritional powder.

[00130] For example, when the nutritional powder is a spray dried nutritional powder, the spray drying step may likewise include any spray drying technique that is known for or otherwise suitable for use in the production of nutritional powders. Many different spray drying methods and techniques are known for use in the nutrition field, all of which are suitable for use in the manufacture of the spray dried nutritional powders herein.

[00131] One method of preparing the spray dried nutritional powder comprises forming and homogenizing an aqueous slurry or liquid comprising predigested fat, and optionally protein, carbohydrate, and other sources of fat as described above, and then spray drying the slurry or liquid to produce a spray dried nutritional powder. The method may further comprise the step of spray drying, drymixing, or otherwise adding additional nutritional ingredients, including any one or more of the ingredients described herein, to the spray dried nutritional powder.

[00132] Other suitable methods for making nutritional compositions are described, for example, in U.S. Pat. No. 6,365,218 (Borschel, etal. U.S. Patent No. 6,589,576 (Borschel, et al.'), U.S. Pat. No. 6,306,908 (Carlson, etal. U.S. Patent Application No. 20030118703 Al (Nguyen, et al.), which descriptions are incorporated herein by reference to the extent that they are consistent herewith.

Methods of Use

[00133] The methods of use according to the general inventive concepts include the oral administration of the nutritional compositions (e.g., infant formulas) that include HMOs, bovine immunoglobulin-containing protein component and, in certain aspects of the present disclosure, combined with PUFAs, nucleotides, and carotenoids to improve or modulate at least one of the conditions or diseases discussed herein, including but not limited to treating or preventing viral infection or symptoms thereof. In certain aspects, symptoms of a viral infection include, but are not limited to, headache, chills, coughing, fussiness and irritability, runny nose, red eyes, otitis media, wheezing, brief apneas, fever, fatigue, decreased appetite, dysphagia, pharyngitis, bronchitis, and pneumonia. In certain aspects, the methods treat, prevent, or ameliorate at least one of the aforementioned symptoms.

[00134] The nutritional compositions as described herein can be administered to individuals including infants generally, or may, in certain aspects of the present disclosure, be administered to a specific subclass of infants that are “in need thereof;” that is, to specific infants that would particularly benefit by administration of the nutritional composition. For example, a specific infant may be “in need of’ the infant formulas as described herein if they are susceptible to (i.e., have one or more of a genetic predisposition, a family history of, and symptoms of the disease or condition) diseases and conditions that can impair/reduce function in one or more of the areas discussed herein, including but not limited to susceptibility to a pathogenic infection (e.g., they are immunocompromised), including a viral infection. Likewise, in another aspect, an adult may be in need of a nutritional composition according to the general inventive concepts for similar reasons.

[00135] Certain specific populations with need include those having substandard or abnormal: maturation and/or development and/or function of the immune response; digestion and/or food tolerance; or maturation and/or development of the immune system; relative to comparable populations.

[00136] Based on the foregoing, because some of the method embodiments according to the general inventive concepts are directed to specific subsets or subclasses of identified individuals (that is, the subset or subclass of individuals “in need” of assistance in addressing one or more specific diseases or specific conditions noted herein), not all individuals will fall within the subset or subclass of individuals as described herein for certain diseases or conditions.

[00137] In certain aspects according to the general inventive concepts, the individual is susceptible to, is infected, or shows signs/symptoms of an infection, including a viral infection. In certain aspects of the present disclosure, the viral infection is due to at least one of Influenza virus (e.g., IAV), Respiratory Syncytial Virus (RSV), Coronavirus (CoV), Rhinovirus (RV), and Adenovirus (AdV). In certain aspects of the present disclosure, the virus is one or more of enveloped and nonenveloped respiratory viruses with DNA or RNA genomes. The virus may be enveloped (e.g. RSV, CoV), nonenveloped (e.g. RV, AdV), and have a genome comprised of DNA (AdV), positive sense RNA (CoV, RV), or negative sense RNA (RSV). In certain aspects of the present disclosure, the virus may be enveloped respiratory virus with negative-sense RNA genome (RSV). In certain aspects of the present disclosure, the virus may be enveloped respiratory virus with positive-sense RNA genome (CoV). In certain aspects of the present disclosure, the virus may be nonenveloped respiratory virus with positive sense RNA genome (RV). In certain aspects of the present disclosure, the virus may be nonenveloped respiratory virus with DNA genome (AdV). In certain exemplary aspects of the present disclosure, administration of the nutritional compositions containing HMOs/ bovine immunoglobulin- containing protein component according to the general inventive concepts treats, prevents, or reduces at least one symptom of a viral infection, including but not limited to a reduction in viral replication.

[00138] When administered to the individual in need thereof, the individual desirably consumes at least one serving of the nutritional composition daily, and in some embodiments, may consume two, three, or even more servings per day. Each serving is desirably administered as a single, undivided dose, although the serving may also be divided into two or more partial or divided servings to be taken at two or more times during the day. The methods according to the general inventive concepts include continuous day after day administration, as well as periodic or limited administration, although continuous day after day administration is generally desirable. The methods according to the general inventive concepts are preferably applied on a daily basis, wherein the daily administration is maintained continuously for at least 3 days, including at least 5 days, including at least 1 month, including at least 6 weeks, including at least 8 weeks, including at least 2 months, including at least 6 months, desirably for at least about 18-24 months, desirably as a long term, continuous, daily, dietary source or supplement.

Examples

[00139] The following examples illustrate exemplary embodiments and/or features of the methods and nutritional compositions according to the general inventive concepts. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the general inventive concepts, as many variations thereof are possible without departing from the spirit and scope of the general inventive concepts. All exemplified amounts are weight percentages based upon the total weight of the composition, unless otherwise specified.

[00140] A nutritional composition comprising an HMO component (e.g., a neutral oligosaccharide and an acidic oligosaccharide) and a bovine immunoglobulin-containing protein component, alone or in combination with another HMO and/or one or more of LCPUFAs, antioxidants, and nucleotides is administered to an individual. The nutritional composition promotes an immune response in the individual (e.g., treating insufficient immune response to pathogens or treating insufficient immune factors). Such a response is demonstrated by a treatment of an infection (e.g., reduction in viral activity or replication), reduction or amelioration in symptoms, modulation of cytokine activity, modulation of inflammation, increase in antibody concentration or specificity, recruitment of immune factors, or one or more other markers of immune response.

[00141] Example 1 : The following is an exemplary powdered nutritional composition (infant formula) according to the general inventive concepts.

[00142] Example 2: The following is an exemplary liquid nutritional composition (infant formula) according to the general inventive concepts.

[00143] Example 3: The following is an exemplary liquid nutritional composition according to the general inventive concepts.

[00144] Example 4: MDCK cells were infected one day post plating, when the cells were sub -confluent. Prior to infection, cells were washed twice with MDCK Influenza Infection Media (Minimum Essential Medium Eagle, 25 mM HEPES, 1% L-glutamine, 1% sodium pyruvate, 0.15% sodium bicarbonate, IX penicillin/streptomycin and 3 pg/mL TPCK-treated Trypsin). The viral solution was prepared by adding 100 pL of IAV to 24 mL of infection media, and then 12 mL of the viral solution was added to each flask of cells. Cells were incubated at 37°C and 5% CO2, and 2 days post infection, the MDCK cell monolayer was completely disrupted by viral infection. Flasks were scraped to remove any remaining cells, and the resulting mixture of cells and supernatant was collected, pooled and clarified by centrifugation (1000 x g for 10 min). Supernatant was collected, aliquoted and stored at -80°C. IAV titer was determined by plaque assay. The influenza A/Hong Kong/2/1968 was used at an MOI of 0.001 for infection of lung epithelial cells. (Description modified from: Duska-McEwen, G., Senft, A.P., Ruetschilling, T.L., Barrett, E.G. and Buck, R.H. (2014) Human Milk Oligosaccharides Enhance Innate Immunity to Respiratory Syncytial Virus and Influenza in Vitro. Food and Nutrition Sciences, 5, 1383-1395. http://dx.doi.org/10.4236/fns.2014.514151).

[00145] Ingredient compositions: HMO blend: The HMO blend is reconstituted from a premade mixture of 2’FL, 3FL, LNT, 3’SL, and 6’SL present in a 52: 13:26:4:5 ratio. It is reconstituted and diluted in infection media to 100 mg/mL and syringe filtered through a 0.2 um filter. Bovine Immunoglobulin G (blgG) was obtained from MP Biomedicals (Catalog No. 641402). ANWP3 is a whey protein consisting of 33% protein by weight.

[00146] The ANWP3 was prepared according to the following process: Starting milk protein source (whey or skimmed milk) were defatted and decaseinated using a 0.14pm ceramic microfiltration (MF) membrane at 50°C to remove bacteria, casein, and fat. The resulting whey permeate stream was subjected to Ultrafiltration (UF) using an 800kDa polymeric membrane at 10°C to enrich Immunoglobulins in final retentate stream. The protein profile of the sample at each step was analyzed by reducing SDS-PAGE and HPLC methods to confirm IgG at higher transmission levels in the final fractions was achieved. The resulting IgG enriched whey streams received a pasteurization heat treatment (72°C x 15s) and subsequently spray dried using a Niro single stage dryer (inlet temp. 185°C, outlet temp. 85°C) to give an IgG enriched whey powder.

[00147] The working concentration of ANWP3 in this assay was 17.2g powder per liter. ANWP3 was measured to contain 5.8% IgG by weight by HPLC. At the reconstitution rate, the final concentration of IgG is Ig/L. Quantitation of IgG differed between HPLC and ELISA measurement. The Ig/L depicted in the figures is based on HPLC measurement. When measured by ELISA, the apparent concentration of IgG falls to 0.606 g/L (based on 3.5% IgG in starting material). While IgA was not measured by HPLC it was measured using ELISA. The concentration of IgA in the test article is 0.411 g/L (based on 2.39% IgA in starting material). Figure 1, which used the immunoglobulin-containing protein component resulting from the process to enrich the IgG concentration described above in combination with HMOs shows good ability to fight influenza A.

[00148] Example 5: For RSV infections, Hep 2 cells were plated in growth media at sufficient concentration to achieve 95% confluence after 24 hours preincubation. Growth media was removed and replaced with OptiMEM containing appropriate concentrations of bAbs and HMO. RSV was added to an MOI of 0.1 and incubated at 37°C for 1 hour. After adsorption, final volume was adjusted to 0.5 mL per well with OptiMEM containing appropriate concentrations of bAbs and HMO. Cells were incubated for two days after which media was harvested for virus titration.

[00149] Hep2 cells were grown for 24 hours in growth media to 95% confluence. Growth media was replaced with OptiMEM and inoculated with dilutions of media from the treatment wells. After 1 hour of adsorption, cells overlaid with growth media containing 0.75% methylcellulose and allowed to develop individual foci. After 5 days of foci development, cells were formalin fixed, immunostained, and counted (Figure 2). Surprisingly, treated whey appears to outperform pasteurized skim and both appear to outperform raw milk on a fixed IgG basis. Furthermore, heat treatments appear to enhance not diminish this activity [00150] Starting milk protein fractions were subjected to ultrafiltration (at a temperature of below about 10 °C) and varying volume concentration factor (VCF) to determine the ratio of milk fraction ingredients in the retentate versus the permeate. The following Table summarizes the results.

[00151] As can be seen from the table, the 800 kDa filtration accompanied with a VCF of greater than 5 provided the bovine-immunoglobulin containing protein component with the best ratio of IgG retention coupled with P-lactoglobulin reduction.

[00152] Unless otherwise indicated herein, all sub-embodiments and optional embodiments are respective sub-embodiments and optional embodiments to all embodiments described herein. While the present application has been illustrated by the description of embodiments thereof, and while the embodiments have been described in considerable detail, it is not the intention of the applicants to restrict or in any way limit the scope of the appended claims to such detail. Additional advantages and modifications will readily appear to those skilled in the art. Therefore, the application, in its broader aspects, is not limited to the specific details, the representative compositions or formulations, and illustrative examples shown and described. Accordingly, departures may be made from such details without departing from the spirit or scope of the applicant’s general disclosure herein.

Claims

WHAT IS CLAIMED IS:

1. A method of treating or preventing a viral infection comprising administering a nutritional composition comprising a bovine immunoglobulin-containing protein component and a human milk oligosaccharide component to the individual, wherein the human milk oligosaccharide (HMO) component comprises:

(a) at least one neutral HMO, and

(b) at least one acidic HMO.

2. The method of claim 1, wherein the bovine immunoglobulin component is selected from IgA, IgG, and IgM.

3. The method of claim 2, wherein the bovine immunoglobulin-containing protein component has been subjected to a process to enrich at least one immunoglobulin above a level in untreated bovine protein.

4. The method of any preceding claim wherein the bovine immunoglobulin- containing protein component comprises IgA.

5. The method of any preceding claim, wherein the bovine immunoglobulin- containing protein component comprises IgA, IgG, and IgM.

6. The method of any preceding claim, wherein the human milk oligosaccharide component is present in an amount of 0.01 mg/mL to 20 mg/mL.

7. The method of any preceding claim, wherein the HMO component comprises at least one N-acetylated HMO.

8. The method of any preceding claim, wherein the HMO component comprises at least one fucosylated HMO.

9. The method of any preceding claim, wherein the at least one neutral HMO comprises 2’-fucosyllactose, lacto-N-tetraose, or both.

10. The method of any preceding claim, wherein the at least one neutral HMO comprises 2’-fucosyllactose.

11. The method of any preceding claim, wherein the at least one neutral HMO comprises lacto-N-tetraose.

12. The method of any preceding claim, wherein the at least one acidic HMO is 6’- sialyllactose.

13. The method of any preceding claim, wherein the HMO component comprises at least one fucosylated HMO, at least one sialylated HMO, and at least one N-acetylated HMO.

14. The method of any preceding claim, wherein the HMO component comprises 2’- fucosyllactose, 6’-sialylactose, and lacto-N-tetraose.

15. The method of any preceding claim, wherein the HMO component comprises 2’FL, 3FL, LNT, 3’SL, and 6’SL.

16. The method of any preceding claim, wherein the HMO component comprises 2’FL, 3 FL, LNT, 3’SL, and 6’SL and is substantially free of other HMOs.

17. The method of any preceding claim, wherein the HMO component comprises 2’FL, 3 FL, LNT, 3’SL, and 6’SL in a weight ratio of approximately (50-55) : (10-15) : (20-30) : (2-7) : (2-7).

18. The method of any preceding claim, wherein the viral infection is a respiratory viral infection.

19. The method of any preceding claims wherein the bovine immunoglobulin- containing protein component has been subjected to ultrafiltration and/or heat treatment.

20. The method of claim 19 wherein the bovine immunoglobulin-containing protein component provides bovine immunoglobulins in an amount of from about 0.01 mg/mL to about 100 mg/mL.

21. The method of claim 19 wherein the bovine immunoglobulin-containing protein component provides bovine immunoglobulin in an amount of from about 0.5 mg/mL to about 10 mg/mL.

22. The method of any one of claims 19-21, wherein the bovine immunoglobulin comprises from 10% to 50% IgA.

23. The method of any preceding claim, wherein the human milk oligosaccharide and bovine immunoglobulin are present in the nutritional composition in a weight ratio of 1 :20 to 20: 1.

24. The method of any preceding claim wherein administering the nutritional composition reduces or ameliorates at least one of the following symptoms: headache, chills, coughing, fussiness and irritability, runny nose, red eyes, otitis media, wheezing, brief apneas, fever, fatigue, decreased appetite, dysphagia, pharyngitis, bronchitis, and pneumonia.

25. A method of preparing a bovine immunoglobulin-containing protein component, comprising: subjecting a milk fraction comprising skim milk and/or whey to microfiltration to defat and decaseinate the milk fraction and provide a sterile retenate, subjecting the defatted and decaseinated milk fraction to a first ultrafiltration step and a volume concentration factor (VCF) of at least about 5, and subjecting the concentrated milk fraction to a second ultrafiltration step to remove lactose and mineral content.

26. The method of claim 25, wherein the first ultrafiltration step comprises a 700-800 kDa membrane ultrafiltration.

27. The method of claim 25, wherein the second ultrafiltration step comprises 10 kDa membrane ultrafiltration.

28. The method of claim 25, wherein the microfiltration is performed with heating to a temperature of from about 30°C to about 50°C.

29. The method of claim 28, wherein the microfiltration step comprises a VCF of at least about 5.

30. The method of claim 25, wherein at least one of the first ultrafiltration step and the second ultrafiltration step is performed at a temperature of about 5°C to about 20°C.

31. The method of claim 25, wherein at least one of the first ultrafiltration step and the second ultrafiltration step comprises a VCF of at least about 5.