RU2809650C2 - Food butyrate for treatment or prevention of allergic disorder - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to the use of one or more compounds having formula (1), (2), (3) or (4) for the treatment or prevention of an allergic disorder. In formulas (1)-(4), R¹, R², R³, R⁴, R⁵ and R⁶ independently represent a long-chain fatty acid containing from 16 to 20 carbon atoms. The invention also relates to the use of a composition containing one or more compounds having formula (1), (2), (3) or (4) for the treatment or prevention of an allergic disorder.

EFFECT: these compounds and compositions containing them have improved organoleptic properties.

19 cl, 6 dwg, 3 tbl, 6 ex

Description

Scope of the invention

The present invention relates to a food source of butyrate having improved organoleptic properties for use in the treatment or prevention of an allergic disorder.

Prerequisites for creating the invention

Allergic diseases are now recognized as epidemic by the World Health Organization (WHO). In Europe and North America, food allergies are estimated to affect approximately 5% of adults and 8% of children (Sicherer, S.H. and Sampson, H.A., 2014. Journal of Allergy and Clinical Immunology, 133(2), pp.291-307). The prevalence of food allergies and anaphylaxis appears to be on the rise, with the greatest increases observed in infants with food allergies or atopic eczema (Koplin, J.J., et al., 2015. Current opinion in allergy and clinical immunology, 15(5), pp .409-416).

Changes in the composition of the intestinal microflora have been proposed as one explanation for the increased incidence of allergic diseases. The gut microbiota activates regulatory T cells (Tregs), which have previously been shown to play a key role in maintaining immune tolerance to allergens (Rivas, M.N. and Chatila, T.A., 2016. Journal of Allergy and Clinical Immunology, 138(3), pp.639-652). In particular, dietary fiber-derived metabolites contribute to intestinal homeostasis and Treg cell biology (Tan, J., et al., 2016. Cell reports, 15(12), pp.2809-2824). Butyric acid is one of the most abundant short-chain fatty acids (SCFAs) produced by the human gut microbiota in response to indigestible dietary fiber. Salts and esters of butyric acid are known as butyrates or butanoates.

Preclinical data showed that direct oral consumption of butyrate was sufficient to prevent the development of food allergies (Tan, J., et al., 2016. Cell reports, 15(12), pp.2809-2824) and alleviate the symptoms of allergic pneumonia (Thio , C. L. P., et al. 2018. Journal of Allergy and Clinical Immunology, 142(6), pp.1867-1883.e12). In addition, elevated levels of fecal butyrate have been associated with accelerated outgrowth of cow's milk allergy in infants (Canani, R.B., et al., 2016. ISME journal, 10(3), p.742), and in addition, higher The abundance of butyrate-producing gut microbiota was associated with poorer symptoms in infants with atopic eczema (Nylund, L., et al., 2015. Allergy, 70(2), pp.241-244). Therefore, butyrate is a strong candidate for modulating food allergic responses through stimulation of Treg cells.

In addition to its established effects on Treg responses, butyrate has also been shown to influence mast cell activity. Mast cells are believed to have important proinflammatory functions as well as potential immunoregulatory functions in various immune disorders through the release of mediators such as histamine, leukotrienes, cytokines, chemokines and neutral proteases (Amin, K., 2012. Respiratory medicine, 106(1) ), pp.9-14). The murine mast cell line MC/9 shows reduced proliferation when exposed to butyrate (Galli, S.J., et al., 1982. The Journal of cell biology, 95(2), pp.435-444), and further studies showed that butyrate inhibits activation of mast cells, inhibiting the release of mediators and the production of the pro-inflammatory cytokine TNFα in mast cells (Diakos, C., et al., 2006. Biochemical and biophysical research communications, 349(2), pp.863-868). Butyrate also effectively inhibited mast cell activation and the production of inflammatory mediators in vivo (Wang, C.C., et al., 2018. Innate immunity, 24(1), pp.40-46).

Antigen-specific stimulation of B cells in the presence of butyrate switched plasmacytic differentiation to the development of regulatory B cells (Breg) and IL-10 production, and Breg induction in vivo was associated with a concomitant reduction in the number of intestinal mast cells after experimental food allergy (Shi, Y ., et al., 2015. Scientific reports, 5, p.17651). Butyrate may also influence mast cell responses by affecting the activation and/or cytokine response of innate lymphoid cells type 2 (ILC2). Butyrate, but not acetate or propionate, was sufficient to alleviate ILC2 cell-induced allergic inflammation (including eosinophilic inflammation) and to reduce cytokine production in human ILC2 cells (Thio, C.L.P., et al. 2018. Journal of Allergy and Clinical Immunology, 142( 6), pp.1867-1883.e12). ILC2 cell responses contributed to the stimulation of IgE and mast cell-mediated experimental food allergy (Chen, C.Y., et al., 2015. Immunity, 43(4), pp.788-802).

Common sources of butyrate are butyric acid and tributyrin, a triglyceride consisting of three ester functional groups with three butyrate moieties and a glycerol backbone. Butyric acid and tributyrin are food additives considered generally generally recognized as safe (GRAS) (21CFR582.60 and 21CFR184.1903, respectively) and are natural components of many dairy products. However, butyric acid is associated with negative sensory properties, such as vomit, feces and cheese odors. Tributyrin also has negative organoleptic properties, in particular, high bitterness. These unpleasant taste and olfactory characteristics may make oral administration of compositions containing these compounds difficult, especially for children. Butyrate components from milk cannot be fortified and thus would require consuming significant amounts of milk fat, which is not practical for practical and nutritional reasons, not least because it would lead to the consumption of large amounts of unwanted calories from animal fat .

Accordingly, it would be beneficial to provide a food source of butyrate having improved sensory properties compared to available solutions for use in the treatment or prevention of an allergic disorder.

Summary of the invention

The present invention provides butyrate source compounds having improved organoleptic properties for use in the treatment or prevention of an allergic disorder. In particular, the compounds have an improved odor and/or taste compared to butyric acid, butyrate salts and/or tributyrin. These compounds can be used as a food source of butyric acid. These compounds can be used, for example, in nutritional compositions, dietary supplements, infant formulas, infant formulas, beverages and pet care products.

According to a first aspect of the present invention, there is provided a compound having the formula

or a combination for use in the treatment or prevention of an allergic disorder, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently a long chain fatty acid containing from 16 to 20 carbon atoms.

According to another aspect of the present invention, there is provided a composition containing a compound having formula (1), (2), (3) or (4), or combinations thereof, for use in the treatment or prevention of an allergic disorder, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently long chain fatty acids having from 16 to 20 carbon atoms.

In one embodiment, the composition contains a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

The composition may contain a compound of formula (1) and a compound of formula (2).

The composition may contain a compound of formula (1) and a compound of formula (3).

The composition may contain a compound of formula (1) and a compound of formula (4).

The composition may contain a compound of formula (2) and a compound of formula (3).

The composition may contain a compound of formula (2) and a compound of formula (4).

The composition may contain a compound of formula (3) and a compound of formula (4).

The composition may contain a compound having formula (1), a compound having formula (2), and a compound having formula (3).

The composition may contain a compound having formula (1), a compound having formula (2), and a compound having formula (4).

The composition may contain a compound having formula (1), a compound having formula (3), and a compound having formula (4).

The composition may contain a compound having formula (2), a compound having formula (3), and a compound having formula (4).

The composition may contain a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

In one embodiment, the compounds having formulas (1), (2), (3) and (4) constitute at least 50 wt.%, 60 wt.%, 70 wt.%, 80 wt.%, 90 wt. .%, 95 wt.% or 99 wt.% of the total triglyceride content in the composition.

In one embodiment, the compounds having formulas (1), (2), (3) and (4) constitute at least 50 wt.%, 60 wt.%, 70 wt.%, 80 wt.%, 90 wt. .%, 95 wt.% or 99 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the compound having formula (4) is the main triglyceride containing a butyrate moiety in the composition.

In one embodiment, the compound of formula (4) constitutes at least 20 wt%, at least 30 wt%, at least 40 wt%, at least 50 wt%, or at least 60 wt%, at least 70 wt.%, at least 80 wt.% or at least 90 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the composition contains a compound of formula (1) and a compound of formula (4), and the combination of a compound having formula (1) and a compound having formula (4) is present in an amount of at least 30%, 40%, 50 %, 60%, 70%, 80% or 90% by weight of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, tributyrin constitutes less than 10% by weight of the total triglyceride content of the composition, preferably less than 8% by weight, more preferably less than 5% by weight of the total triglyceride content of the composition.

In one embodiment, the composition further comprises vitamin A and/or dietary fiber and/or a probiotic.

In one embodiment, the composition is a nutritional composition, wherein preferably the nutritional composition is a dietary supplement, infant formula, infant formula, beverage, or pet care product.

In one embodiment, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and/or R ⁶ are an unsaturated fatty acid, preferably monounsaturated.

In one embodiment, ^R1 , ^R2 , ^R3 , ^R4 , ^R5 and/or ^R6 are selected from the group consisting of oleic acid, palmitic acid or linoleic acid, preferably each of ^R1 , ^R2 , ^R3 , R ⁴ , R ⁵ and R ⁶ represent oleic acid.

In one embodiment, the allergic disorder is selected from one or more of the group consisting of food allergy, food intolerance, respiratory allergy, and skin allergy.

In one embodiment, the allergic disorder is selected from one or more of the group consisting of rhinitis, asthma, urticaria or hay fever, allergic conjunctivitis, dermatitis, atopic dermatitis, contact dermatitis, eczema, atopic eczema, urticaria, psoriasis, eosinophilic esophagitis and related eosinophilia gastrointestinal disease, allergic diarrhea, vomiting, abdominal pain or bloating.

In one embodiment, the allergen for an allergic disorder is selected from one or more of a food allergen, dust mite, pollen, mold or mildew spores, weed pollen, tree pollen, grass pollen, fleas, pet hair or other animals, pet feathers or dander .

In one embodiment, the allergen in an allergic disorder is a food allergen, wherein preferably the food allergen is selected from nut, tree nut, peanut, fish, shellfish, crustacean, milk, egg, soy, gluten, grain, wheat, oat, barley, rye, celery, corn, lupine, sulfites, sesame, mustard, rice, poultry and meat.

In one embodiment, the compounds or combinations thereof have improved organoleptic properties compared to butyric acid, tributyrin and/or butyrate salts.

In one embodiment, the compounds are administered to a young or adult mammal, preferably an infant, toddler, juvenile or adult, pet or farm animal.

Brief description of graphic materials

In fig. 1 is a graphical representation of the BLG-specific immunoglobulin E (IgE) response in mice sensitized with the milk protein β-lactoglobulin (BLG) and in control mice.

In fig. 2 is a graphical representation of the BLG-specific immunoglobulin G1 (IgG1) response in mice sensitized with the milk protein β-lactoglobulin (BLG) and in control mice.

In fig. 3 is a graphical representation of the cumulative clinical score after oral challenge with OVA in mice of the “control group” (negative control), “allergic group 1” (BLG-sensitized positive control); "allergic group 2, receiving sodium butyrate" and "allergic group 3, receiving attractive-tasting sodium butyrate."

In fig. 4 is a graphical representation of the OVA-specific IgE response to oral OVA challenge in allergic and control mice: “control group” (negative control), “allergic group 1” (BLG-sensitized positive control); "allergic group 2, receiving sodium butyrate" and "allergic group 3, receiving attractive-tasting sodium butyrate."

In fig. 5 is a graphical representation of the OVA-specific IgG1 response to oral OVA challenge in allergic and control mice: “control group” (negative control), “allergic group 1” (BLG-sensitized positive control); "allergic group 2, receiving sodium butyrate" and "allergic group 3, receiving attractive-tasting sodium butyrate."

In fig. 6 is a graphical representation of the mast cell-specific protease 1 (MCPT-1) response to oral OVA challenge in allergic and control mice: “control group” (negative control), “allergic group 1” (BLG-sensitized positive control); "allergic group 2, receiving sodium butyrate" and "allergic group 3, receiving attractive-tasting sodium butyrate."

Detailed Description of the Invention

As used herein, the terms “comprising,” “comprising,” and “consisting of” are synonymous with the terms “including” or “includes”; or “comprising” or “comprising”, and they are inclusive, or non-limiting, and do not exclude additional non-added terms, elements or stages. The terms “comprising”, “comprising” and “consisting of” also include the term “comprising”.

Triglycerides

A triglyceride (also known as triacylglycerol) is a triester made from glycerol and three fatty acids. Under conditions of hydrolysis, such as during digestion, triglycerides can be a source of fatty acids. For example, tributyrin potentially provides three moles of butyric acid per mole of tributyrin.

Fatty acids are carboxylic acids with a long tail (chain). Fatty acids can be unsaturated or saturated. Fatty acids that are not attached to other molecules are called free fatty acids (FFA).

The term "fatty acid moiety" refers to the portion of a triglyceride that is produced from a fatty acid by esterification reaction with glycerol. The triglycerides used in the present invention contain at least one butyric acid residue and at least one long-chain fatty acid residue.

Preferred long chain fatty acids for use in the present invention are fatty acids having from 16 to 20 carbon atoms. Examples of long chain fatty acids include oleic acid, palmitic acid, stearic acid and linoleic acid. Preferably the long chain fatty acid is oleic acid. For example, the present invention provides a compound having the formula

or combinations thereof for use in the treatment or prevention of an allergic disorder.

Other examples of triglycerides that can be used in the present invention include: 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2 -oleoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, 1-oleoyl-2-butyryl-3-linoleoylglycerol, 1-linoleoyl-2-butyryl -3-oleoylglycerol, 1-butyryl-2-linoleoyl-3-oleoylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3 -butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, 1-stearoyl-2-oleoyl-3-butyrylglycerol, 1,2-dioleoyl-3-palmitoylglycerol, 1-palmitoyl-2,3-dioleoylglycerol, 1,2 -dioleoyl-3-linoleoylglycerol and 1-linoleoyl-2,3-dioleoylglycerol.

The triglycerides of the present invention can be synthesized, for example, by esterifying long-chain fatty acid(s) and butyric acid with glycerol.

The triglycerides of the present invention can be synthesized, for example, by transesterification between tributyrin and another triglyceride containing long chain fatty acids. In one embodiment, the source of long chain fatty acids is high oleic sunflower oil. This leads to the formation of triglycerides containing predominantly butyrate and oleate functional groups. The compounds are dairy-free, cholesterol-free, and vegan-friendly. Fatty acids are released from triglycerides by lipases that are naturally present in the gastrointestinal tract. Compared to butyrate salts, the compounds do not add additional mineral salts to the final composition.

Alternative methods for synthesizing triglycerides can be practiced by one skilled in the art.

As an example, a method for producing 1,3-dibutyryl-2-palmitoylglycerol (BPB) is shown below:

In another example, triglycerides can be synthesized by esterifying a long-chain fatty acid monoacylglycerol (MAG) with butyric acid (BA).

For example, by esterifying a long-chain fatty acid monoacylglycerol (MAG) with butyric acid (BA), removing water. Below, as an example, is shown a method for producing 1,2-dibutyryl-3-oleoylglycerol:

The esterification reaction is preferably carried out at a molar ratio of butyric acid (BA) : monoacylglycerol (MAG) ≥ 2; those. with a molar excess of butyric acid. Removal of water can be carried out by conventional methods commonly used in the art.

In the present invention, a triglyceride containing one butyrate residue can be used. In an alternative embodiment, a mixture of different triglycerides containing butyrate moieties can be used.

The triglycerides can be further subjected to decolorization and/or deodorization steps conventional in the art and well known to one skilled in the art. An example would be the steps traditionally used in the production of vegetable oils.

Compositions

The compounds of the present invention can be administered in the form of a composition. Thus, the present invention provides compositions comprising the butyrate moiety-containing triglycerides mentioned herein for use in the treatment or prevention of an allergic disorder.

In one embodiment, the composition as defined herein contains a combination of a compound having formula (1) and a compound having formula (2).

In one embodiment, the compound of formula (1) is present in an amount of at least 10% by weight of the total triglyceride content of the composition, and the compound of formula (2) is present in an amount of at least 10% by weight of the total triglyceride content of the composition.

In one embodiment, the compound having formula (1) is present in an amount of at least 15 wt.% of the total triglyceride content in the composition, and the compound having formula (2) is present in an amount of at least 15 wt.% of the total triglyceride content in the composition.

In one embodiment, the compound having formula (1) is present in an amount of at least 20 wt.% of the total triglyceride content in the composition, and the compound having formula (2) is present in an amount of at least 20 wt.% of the total triglyceride content in the composition.

In one embodiment, the compound having formula (1) is present in an amount of at least 20% by weight of the total triglyceride content of the composition, and the compound having formula (2) is present in an amount of at least 30% by weight of the total triglyceride content in the composition.

In one embodiment, the compound having formula (1) constitutes from about 20 wt.% to about 40 wt.% of the total triglyceride content of the composition, and/or the compound having formula (2) constitutes from about 30 wt.% up to about 40 wt.% of the total triglyceride content in the composition.

In one embodiment, the compound of formula (1) and the compound of formula (2) constitute at least 20 wt%, 30 wt%, 40 wt%, 50 wt%, 60 wt%, or 70 wt% of the total the triglyceride content of the composition, preferably from about 40 wt.% to about 80 wt.% or from about 50 wt.% to about 75 wt.% of the total triglyceride content in the composition.

In one embodiment, the composition further comprises a compound of formula (3), wherein preferably the compound of formula (3) constitutes at least 2 wt%, 3 wt%, 4 wt% or 5 wt% of the total triglyceride content of the composition; and/or the composition further comprises a compound having formula (4), wherein preferably the compound having formula (4) constitutes at least 1 wt%, 2 wt% or 3 wt% of the total triglyceride content of the composition.

In one embodiment, the compound having formula (1) is present in an amount of at least 20 wt.% of the total butyric acid-containing triglycerides in the composition, and the compound having formula (2) is present in an amount of at least 30 wt. % of the total content of butyric acid-containing triglycerides in the composition.

In one embodiment, the compound having formula (1) constitutes from about 30 wt.% to about 50 wt.% of the total butyric acid-containing triglycerides in the composition, and/or the compound having formula (2) represents from about 40 wt.% wt.% to about 60 wt.% of the total content of butyric acid-containing triglycerides in the composition.

In one embodiment, the compound having formula (1) and the compound having formula (2) constitute at least 20%, 30%, 40%, 50%, 60%, 70% or 80% by weight of the total content butyric acid-containing triglycerides in the composition, preferably from about 60% to about 90% by weight of the total butyric acid-containing triglycerides in the composition.

In one embodiment, the compound having formula (4) is the main triglyceride containing a butyrate moiety in the composition.

In one embodiment, the compound of formula (4) constitutes at least 20 wt%, at least 30 wt%, at least 40 wt%, at least 50 wt%, or at least 60 wt%, at least 70 wt.%, at least 80 wt.% or at least 90 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the composition contains a compound of formula (1) and a compound of formula (4), and the combination of a compound having formula (1) and a compound having formula (4) is present in an amount of at least 30%, 40%, 50 %, 60%, 70%, 80% or 90% by weight of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the compound of formula (5) constitutes at least 10% by weight of the total triglyceride content of the composition, and/or the compound of formula (6) constitutes at least 10% by weight of the total triglyceride content of the composition.

In one embodiment, the compound of formula (5) constitutes at least 15% by weight of the total triglyceride content of the composition, and/or the compound of formula (6) constitutes at least 15% by weight of the total triglyceride content of the composition.

In one embodiment, the compound of formula (5) constitutes at least 15% by weight of the total triglyceride content of the composition, and/or the compound of formula (6) constitutes at least 20% by weight of the total triglyceride content of the composition.

In one embodiment, the compound of formula (5) constitutes at least 20% by weight of the total triglyceride content of the composition, and/or the compound of formula (6) constitutes at least 20% by weight of the total triglyceride content of the composition.

In one embodiment, the compound having formula (5) constitutes from about 15 wt.% to about 30 wt.% of the total triglyceride content of the composition, and/or the compound having formula (6) constitutes from about 20 wt.% up to about 30 wt.% of the total triglyceride content in the composition.

In one embodiment, the compound of formula (5) and the compound of formula (6) comprise at least 20 wt%, 30 wt%, or 40 wt% of the total triglyceride content of the composition, preferably from about 30 wt% to about 60 wt%. wt.%, or from about 40 wt.% to about 50 wt.% of the total triglyceride content in the composition.

In one embodiment, the composition further comprises a compound of formula (7), preferably the compound of formula (7) constitutes at least 2% by weight or 3% by weight of the total triglyceride content of the composition; and/or the composition further contains a compound having formula (8), and preferably the compound having formula (8) constitutes at least 2 wt.% or 3 wt.% of the total triglyceride content in the composition.

In one embodiment, the compound having formula (5) constitutes at least 10% by weight of the total butyrate moiety-containing triglycerides in the composition, and the compound having formula (6) constitutes at least 10% by weight of the total content of triglycerides containing a butyrate residue in the composition.

In one embodiment, the compound having formula (5) constitutes at least 15 wt.% of the total butyrate moiety-containing triglycerides in the composition, and the compound having formula (6) constitutes at least 15 wt.% of the total content of triglycerides containing a butyrate residue in the composition.

In one embodiment, the compound having formula (5) constitutes at least 15 wt.% of the total butyrate moiety-containing triglycerides in the composition, and the compound having formula (6) constitutes at least 20 wt.% of the total content of triglycerides containing a butyrate residue in the composition.

In one embodiment, the compound having formula (5) constitutes at least 20% by weight of the total butyrate moiety-containing triglycerides in the composition, and the compound having formula (6) constitutes at least 20% by weight of the the total content of triglycerides containing a butyrate residue in the composition.

In one embodiment, the compound having formula (5) constitutes from about 15 wt.% to about 30 wt.% of the total triglyceride containing butyrate residue in the composition, and the compound having formula (6) represents from about 20 wt.% to about 30 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the composition further comprises a compound having formula (7), preferably the compound having formula (7) constitutes at least 2 wt.% or 3 wt.% of the total content of triglycerides containing butyrate residue in the composition; and/or the composition further contains a compound having formula (8), and preferably the compound having formula (8) constitutes at least 2 wt.% or 3 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In another embodiment, the compound having formula (8) is the main triglyceride containing a butyrate moiety in the composition.

In one embodiment, the compound of formula (8) constitutes at least 20 wt%, at least 30 wt%, at least 40 wt%, at least 50 wt%, or at least 60 wt%, at least 70 wt.%, at least 80 wt.% or at least 90 wt.% of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the compound having formula (8) constitutes from about 20% to about 95% by weight of the total butyrate moiety-containing triglycerides in the composition, for example, from about 30% to about 90%, or about 40% to about 80% by weight of the total butyrate-containing triglycerides in the composition, for example, from about 50% to about 70% by weight of the total butyrate-containing triglycerides in the composition.

In one embodiment, the compound having formula (8) constitutes from about 50% to about 90% by weight of the total butyrate moiety-containing triglycerides in the composition, for example, from about 60% to about 80% by weight of the total content triglycerides containing a butyrate residue in the composition.

In one embodiment, the composition contains a compound of formula (8) and a compound of formula (5), and the combination of a compound having formula (8) and a compound having formula (5) is present in an amount of at least 30%, 40%, 50 %, 60%, 70%, 80% or 90% by weight of the total content of triglycerides containing butyrate residue in the composition.

In one embodiment, the composition of the present invention may further comprise 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2-oleoyl- 3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, 1-oleoyl-2-butyryl-3-linoleoylglycerol, 1-linoleoyl-2-butyryl-3- oleoylglycerol, 1-butyryl-2-linoleoyl-3-oleoylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, 1-stearoyl-2-oleoyl-3-butyrylglycerol, 1,2-dioleoyl-3-palmitoylglycerol, 1-palmitoyl-2,3-dioleoylglycerol, 1,2-dioleoyl- 3-linoleoylglycerol and/or 1-linoleoyl-2,3-dioleoylglycerol.

In one embodiment, tributyrin constitutes less than 10% by weight of the total butyrate-containing triglycerides in the composition, preferably less than 8% by weight, more preferably less than 5% by weight of the total butyrate-containing triglycerides in the composition.

The composition of the present invention may be presented, for example, in dry (eg powder), liquid or jelly form.

The composition of the present invention may be presented, for example, in the form of a tablet, dragee, capsule, gelatin capsule, powder, granule, solution, emulsion, suspension, coated particle, spray-dried particle or tablet.

The composition may be in the form of a pharmaceutical composition and may contain one or more suitable pharmaceutically acceptable carriers, diluents and/or excipients. Examples of such suitable excipients for the compositions described herein can be found in Handbook of Pharmaceutical Excipients, 2nd Edition, (1994), ed. A Wade and PJ Weller. Suitable carriers or diluents for therapeutic use are also well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985).

As or in addition to a carrier, excipient or diluent, the pharmaceutical compositions may contain any suitable binder(s), lubricant(s), suspending agent(s). e) agent(s), coating agent(s) and/or solubilizing agent(s). Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, bulk lactose, beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, gum tragacanth or sodium alginate, carboxymethylcellulose and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.

Preservatives, stabilizers, coloring agents and even flavoring agents may be included in the composition. Examples of preservatives include sodium benzoate, sorbic acid and p-hydroxybenzoic acid esters. Antioxidants and suspending agents may also be used.

The composition may be a nutritional composition.

The expression “nutritional composition” means a composition that is fed to a subject. The nutritional composition is typically for oral administration and may include a lipid or fat source and a protein source. It may also contain a source of carbohydrates. In one embodiment, the nutritional composition contains only a lipid or fat source. In other specific embodiments, the nutritional composition contains a source of lipids (or fat) in combination with a source of protein and/or a source of carbohydrates.

In some specific embodiments, the nutritional composition of the invention is an “enteral nutritional composition,” meaning that the digestive tract is used to administer the food product. Gastric insertion may involve the use of a tube inserted through the oronasal passage or a tube in the abdomen leading directly into the stomach. In particular, this can be used in hospitals and clinics.

The composition in accordance with the present invention may be a dietary supplement.

The term "dietary supplement" can be used to mean an addition to a subject's diet (typically used alone, but can also be added to any type of composition intended for consumption). It may be presented, for example, in the form of tablets, capsules, lozenges or liquid. The additive may further contain protective hydrocolloids (such as gums, proteins, modified starches), binders, film-forming agents, encapsulating agents/materials, wall/shell materials, matrix compounds, coatings, emulsifiers, surfactants, solubilizing agents (oils, fats, waxes, lecithins, etc.), adsorbents, carriers, fillers, auxiliary compounds, dispersing agents, wetting agents, processing aids (solvents), anti-caking agents, taste-masking agents, weighting agents, gelling agents and gelling agents. The dietary supplement may also contain conventional pharmaceutical additives and excipients, excipients and diluents, including, without limitation, water, gelatin of any origin, vegetable gums, lignin sulfonate, talc, sugars, starch, gum arabic, vegetable oils, polyalkylene glycols, flavoring agents, preservatives, stabilizers, emulsifying agents, buffers, lubricants, dyes, wetting agents, fillers, etc.

When the composition is a supplement, it may be provided in unit dosage form.

The composition in accordance with the invention may be an infant formula (for example, an infant formula), a follow-up formula or a weaning formula, or an infant formula for children from 1 to 3 years of age, an infant food, a cereal-based infant composition, or a fortifier, such as human breast milk fortifier.

The expression “infant formula”, as used herein, refers to a food product specifically intended for use by infants during the first months of life, which in itself satisfies the nutritional needs of this category of persons (e.g. Article 2(c) of the Directive European Commission 91/321/EEC 2006/141/EC of 22 December 2006 on infant formula and formula for complementary feeding).

Typically, initial formula as a breast milk substitute is intended for infants from birth. Formula of the next level or for complementary children, as a rule, begins to be given from the 6th month. It constitutes the main liquid element in the gradually increasing variety of diets for this category of people. "Formula for infants 1 to 3 years" (or GUM) is given from one year of age onwards. Essentially, it is a milk-based drink tailored to the specific nutritional needs of young children.

The term "fortifier" refers to liquid or dry nutritional compositions suitable for mixing with breast milk (human milk) or infant formula. The term "breast milk" should be understood as human milk, or human milk colostrum, or donor milk, or donor milk colostrum.

The composition in accordance with the invention may be a dairy product, a liquid beverage, a powdered beverage, a dehydrated soup, a dietary supplement, a meal replacement, a nutrition bar, a cereal product, a confectionery product, or a dry pet food.

The composition may additionally contain dietary fiber. Dietary fiber has been shown to improve oral tolerance and protect against food allergies (Tan, J., et al., 2016. Cell reports, 15(12), pp.2809-2824). "Dietary fiber" may contain at least one non-digestible oligosaccharide (eg, prebiotics). Prebiotics may be present in an amount from 0.3 to 10% by weight of the composition. Dietary fiber and/or prebiotics may stimulate the production of endogenous butyrate by gut microflora and thus provide additional beneficial effects.

Prebiotics are generally indigestible in the sense that they are not broken down or absorbed in the stomach or small intestine and thus remain in their native form as they pass into the large intestine, where they are selectively fermented by beneficial bacteria. Examples of prebiotics include specific oligosaccharides such as fructooligosaccharides (FOS), inulin, xylooligosaccharides (XOS), polydextrose, or any mixture thereof. In a specific embodiment, the prebiotics may be fructooligosaccharides and/or inulin. In a specific embodiment, the prebiotics are a combination of FOS with inulin, such as the product sold by BENEO-Orafti under the trade name oligofructose Orafti® (formerly Raftilose®), or the product sold by BENEO-Orafti under the trade name inulin Orafti® (formerly Raftiline ®). Another example is the combination of 70% short-chain fructo-oligosaccharides and 30% inulin, trademarked by Nestle under the trademark Prebio 1. The nutritional composition of the invention may also contain at least one milk oligosaccharide, which may be a BMO (bovine milk oligosaccharide) and/or OGM (breast milk oligosaccharide). In a specific embodiment, the nutritional composition in accordance with the present invention contains a mixture of oligosaccharides containing from 0.1 to 4.0 wt.% N-acetylated oligosaccharide(s), from 92.0 to 98.5 wt. % galactooligosaccharide(s) and from 0.3 to 4.0 wt.% sialylated oligosaccharide(s).

The composition of the present invention may further contain at least one probiotic (or probiotic strain), such as a probiotic bacterial strain. Consumption of probiotic strains may also promote endogenous butyrate production by gut microflora and thus provide additional beneficial effects.

The most commonly used probiotic microorganisms are mainly bacteria and yeasts of the following genera: Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium and Saccharomyces.

In some specific embodiments, the probiotic is a probiotic bacterial strain. In some specific embodiments, this is Bifidobacterium and/or Lactobacillus.

The nutritional composition according to the invention may contain from 10e3 to 10e12 CFU of the probiotic strain, more preferably from 10e7 to 10e12 CFU, for example from 10e8 to 10e10 CFU of the probiotic strain per gram of composition based on dry matter weight.

In one embodiment, the probiotics are viable. In another embodiment, the probiotics are non-propagating or inactivated. These may also be parts of probiotics, such as cell wall components or metabolic products of probiotics. In some other embodiments, both viable probiotics and inactivated probiotics can be used. The nutritional composition of the invention may further comprise at least one phage (bacteriophage) or a mixture of phages, preferably directed against pathogenic microorganisms Streptococci, Haemophilus, Moraxella and Staphylococci.

The nutritional composition of the invention, and especially the infant formula, essentially contains a protein source, a carbohydrate source and a lipid source. However, in some embodiments, especially when the nutritional composition of the invention is a supplement or fortifier, only lipids (or a source of lipids) may be present.

The nutritional composition according to the invention may contain a protein source. Protein can be contained in amounts from 1.6 to 3 g per 100 kcal. In some embodiments, particularly when the composition is intended for premature infants/young children, the amount of protein may be from 2.4 to 4 g/100 kcal or greater than 3.6 g/100 kcal. In some other embodiments, the amount of protein may be less than 2.0 g per 100 kcal, such as 1.8 to 2 g/100 kcal, or the protein may be contained in an amount less than 1.8 g per 100 kcal.

Protein sources based on, for example, whey, casein and mixtures thereof, as well as soy-based protein sources can be used. With respect to whey proteins, the protein source may be based on acid whey or sweet whey or mixtures thereof and may include alpha-lactalbumin and beta-lactoglobulin in any desired proportions. In some embodiments, the protein source is predominantly whey (ie, more than 50% of the proteins are derived from whey proteins, e.g., 60%> or 70%>). The proteins may be native or hydrolyzed, or they may be a mixture of native and hydrolyzed proteins. In some embodiments, the protein source may also be provided partially or entirely in the form of added amino acids.

The term “native” means that the main part of the proteins is native, i.e. their molecular structure is unchanged, for example at least 80% of the proteins are unchanged, for example at least 85% of the proteins are unchanged, preferably at least 90% of the proteins are unchanged, even more preferably at least 95% of the proteins are unchanged, for example, at least 98% of proteins are unchanged. In a particular embodiment, 100% of the proteins are unchanged.

The term "hydrolyzed" in the context of the present invention means a protein that has been hydrolyzed or broken down into its constituent amino acids.

Proteins can be fully or partially hydrolyzed. If it is necessary to use hydrolyzed proteins, the hydrolysis process can be carried out as desired and in a manner known in the art. For example, whey protein hydrolysates can be produced by enzymatic hydrolysis of a whey fraction in one or more steps. If the whey fraction used as the starting material is substantially lactose-free, it has been found that protein lysine blocking occurs to a much lesser extent during the hydrolysis process. This allows the degree of lysine blocking to be reduced from about 15 wt% total lysine to less than about 10 wt% > lysine; for example, about 7 wt.% lysine, which significantly improves the nutritional quality of the protein source.

In one specific embodiment, the proteins of the composition are hydrolyzed, substantially hydrolyzed, or partially hydrolyzed. The degree of hydrolysis (DH) of a protein can be from 2 to 20, or from 8 to 40, or from 20 to 60, or from 20 to 80, or more than 10, 20, 40, 60, 80 or 90. For example, Nestle offers nutritional compositions are available for sale under the Peptamen® trademark, containing hydrolysates with a degree of hydrolysis of less than about 15%.

In some embodiments, the protein is substantially hydrolyzed. For example, infant formula products containing highly hydrolyzed protein are available from Nestlé under the brand name Althera®, Alfaré®.

At least 70%, 80%, 85%, 90%, 95% or 97% of the proteins can be hydrolyzed. In a specific embodiment, 100% of the proteins are hydrolyzed.

In one particular embodiment, the proteins are in the form of amino acids. For example, infant formula products based on amino acids as a source of protein are available from Nestlé under the brand name Alfamino®.

In one specific embodiment, the proteins of the composition are a plant protein.

The nutritional composition in accordance with the present invention may contain a source of carbohydrates. This is particularly preferred when the nutritional composition of the invention is an infant formula. In this case, any carbohydrate source commonly found in infant formulas can be used, such as lactose, sucrose, maltodextrin, starch, and mixtures thereof, although lactose is one of the preferred carbohydrate sources in infant formulas.

The nutritional composition of the invention may also contain all the vitamins and minerals considered essential in the daily diet in nutritionally significant amounts. Some vitamins and minerals have minimum requirements. Examples of minerals, vitamins and other nutrients that are optionally present in the composition of the invention include vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid , inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine and L-carnitine. Minerals are usually added in the form of salts. The presence and amounts of specific minerals and other vitamins will vary depending on the intended population.

In one specific embodiment, the composition may further comprise vitamin A and/or retinol. Butyrate can stimulate CD103-expressing dendritic cells (CD103+ DCs) to convert vitamin A into retinoic acid, which promotes the differentiation of naïve T cells into Treg cells (Tan, J., et al., 2016. Cell reports, 15(12), pp .2809-2824). In particular, Tan et al. showed that dietary fiber with vitamin A increased the activity of tolerogenic CD103+ DCs.

If necessary, the nutritional composition of the invention may contain emulsifiers and stabilizers such as soy, lecithin, mono- and diglyceryl esters of citric acid, and the like. The nutritional composition of the invention may also contain other substances that may provide beneficial effects, such as lactoferrin, osteopontin, TGFbeta, sIgA, glutamine, nucleotides, nucleosides and the like.

The nutritional composition according to the invention can be prepared by any suitable method. For example, the composition can be prepared by mixing the components in suitable proportions, optionally mixed with one or more carriers, and then mixing the dry mixed mixture with a diluent to form a liquid mixture. The liquid mixture can then be homogenized, pasteurized and optionally spray dried if the final product is to be a powder. The composition can be homogenized before pasteurization or after pasteurization.

For example, a formula, such as an infant formula, can be prepared by mixing a protein source, a carbohydrate source and a fat source in appropriate proportions. At this stage, emulsifiers (if used) can be included. Vitamins and minerals can be added at this stage, but are usually added later to prevent thermal decomposition. Any lipophilic vitamins, emulsifiers, etc. may be dissolved in the fat source before mixing. Water, preferably purified by reverse osmosis, can then be added to form a liquid mixture. The water temperature conveniently ranges from about 50°C to about 80°C to facilitate dispersion of the ingredients. Commercially available thinners can be used to form a liquid mixture. Any oligosaccharides can be added at this stage, especially if the final product is to be in liquid form. If the final product is to be a powder, these components can also be added at this stage if desired. The liquid mixture is then homogenized, for example, in two stages.

Allergic disorders

The compounds defined herein are a source of butyrate/butyric acid and, therefore, can be used for the prevention or treatment of allergic disorders. The compounds can be used in infants, children or adults.

Of critical importance and greatest interest is allergic sensitivity in childhood, especially in early childhood and especially to food allergens, since the development of an “allergic phenotype” or “atopy” has been demonstrated to contribute to subsequent sensitivity to other allergens. Eczema in infants is also a predisposition factor for later developing food allergies or other types of allergies. Thus, childhood allergies may be the first stage of an allergic cascade leading to multiple allergies later in life. This process is usually called the "atopic march." For example, children with persistent food hypersensitivity early in life have a significantly increased risk of developing allergic rhinitis (hay fever) or asthma later in childhood (Ostblom, E. et al. (2008). Phenotypes of food hypersensitivity and development of allergic diseases during the first 8 years of life, Clinical and Experimental Allergy, 38 (8): 1325-1332). Children with milder forms of food hypersensitivity also have an increased risk of developing respiratory allergies, but to a lesser extent than children with persistent food hypersensitivity. Thus, reducing the severity of food hypersensitivity may be a decisive factor in slowing down the “atopic march.” In this context, treatment of allergy cases and prevention of allergies during childhood and infancy are of utmost importance.

Infants' immune systems actively develop during the first few years of life. Targeting, preventing, preventing, treating, reducing, or modulating allergic reactions in these younger patients may influence their allergic profile in the short term, as well as have a long-term impact later in life. In one embodiment, the prevention or treatment of an allergic disorder is accomplished primarily through prophylaxis. “Primary prevention” is the effect of preventing or reducing the risk of patient sensitivity to allergens, which is characterized by the absence or decrease in concentrations of allergen-specific IgE antibodies. Prevention or reduction of sensitization will result in the absence or reduction of allergic symptoms when exposed to the same allergen. By modulating the mechanism of a patient's sensitivity to one allergen or one group of allergens (primary prevention), it is also possible to modulate the subsequent allergic response. Thus, in one embodiment, preventing or treating an allergic disorder includes reducing or preventing an allergic reaction to subsequent exposure to an allergen or allergens in an allergic individual (an individual with a history of allergies).

Food allergens are among the first allergens that infants encounter early in life: non-exclusively breastfed infants are typically exposed to cow's milk proteins. Dairy proteins are indeed one of the most common causes of food allergies in infants, followed by egg, soy and wheat proteins. In general, in infants and young children, food allergies can manifest as skin (rash, eczema, etc.) and gastrointestinal (abdominal cramps; pain, especially in the abdomen; vomiting) symptoms. Food allergies are the most common trigger for severe allergic reactions, which can lead to life-threatening anaphylaxis.

Additional sensitivities and allergies may also occur when an infant/young child is exposed to a new food such as grains, vegetables, fruits, nuts or fish and is exposed to airborne allergens such as pollen, house dust mites and animal dander. Adults mainly suffer from contact and respiratory allergies. Recent WHO data (Clark, M. J. and. Million, R. P (2009) Allergic rhinitis: market evolution, Nature Reviews, Drug Discovery, 8, p. 271–272) indicate that up to 30–40% of the world population suffers any form of respiratory allergy.

Animals, especially small animals such as pets, and especially companion animals such as dogs and cats, can also suffer from food allergies and food intolerances, as well as reactions to environmental allergens. They typically have symptoms similar to humans, such as gastrointestinal disorders such as diarrhea, vomiting and abdominal discomfort, as well as dermatitis or itching. In small animals, particularly dogs, the most common cause of chronic diarrhea is foodborne enteropathy (food enteropathy or foodborne diarrhea).

By preventing or reducing the risk of sensitization of subjects to allergens, the compounds and compositions of the present invention can be used to prevent or treat food allergies, food intolerances, respiratory allergies and skin allergies.

In some embodiments, the allergic response is a specific IgE-associated immune response and/or a T cell-mediated hypersensitivity reaction. Thus, in some embodiments, reducing or preventing allergies includes reducing or preventing specific IgE-associated immune responses and/or T cell-mediated hypersensitivity reactions. In some embodiments, allergic inflammation is reduced and/or oral tolerance is increased.

As used herein, the term “food allergy” refers to an abnormal immune response to one or more food allergens, typically an IgE response caused by histamine release, but also including immune responses other than IgE. Symptoms of food allergies may include itching, eczema, urticaria, tongue swelling, vomiting, diarrhea, hives, difficulty breathing, or low blood pressure. If symptoms are severe and affect more than one body system, the reaction is called anaphylaxis.

As used herein, the term “food allergen” refers to proteins or derivatives thereof that cause abnormal immune responses. Purified food allergens can be named using the systematic nomenclature of the Subcommittee on Allergen Classification of the World Health Organization and the International Union of Immunological Societies. Allergen names consist of the abbreviated scientific name of the source (genus: 3–4 letters; species: 1–2 letters) and a number in Arabic numerals, for example, Der p 1 for the first described allergen from the house dust mite Dermatophagoides pteronyssinus. Food allergens are derived proteins with various biological functions, including proteases, ligand-binding proteins, structural proteins, pathogen-associated proteins, lipid transfer proteins, profilins, and calcium-binding proteins. A list of food allergens is provided on the official website of the WHO/IUIS Allergen Nomenclature Database, http://www.allergen.org/index.php. (Radauer, C., et al., 2014. Allergy, 69(4), pp.413-419 and Pomés, A., et al., 2018. Molecular immunology).

In one embodiment, the food allergen is selected from one or more of the list consisting of nut, tree nut, peanut, fish, shellfish, crustacean, milk, egg, soy, gluten, grain, wheat, oats, barley, rye, celery, corn , lupine, sulfites, sesame, mustard, rice, poultry and meat.

As used herein, the term "food intolerance" refers to an adverse reaction, often delayed, to a food, drink, food additive or substance found in food that causes symptoms in one or more organs and systems of the body, but generally refers to reactions different from food allergies. The term “food hypersensitivity” can broadly refer to both food intolerances and food allergies.

As used herein, the term “respiratory allergy” refers to an abnormal immune response to one or more airborne allergens. Airborne allergens may include pollen, mold or mold spores, weed pollen, tree pollen, grass pollen and dander. Respiratory allergies may include, for example, allergic rhinitis and allergic asthma. Symptoms of allergic rhinitis (hay fever) include a runny or stuffy nose, sneezing, red eyes, itching, watery eyes and swelling around the eyes. Symptoms of allergic asthma include cases of respiratory obstruction, coughing, chest tightness and respiratory failure.

As used herein, the term “skin allergy” refers to an abnormal immune response caused by contact with one or more environmental allergens or consumption of an allergenic food. Environmental allergens may include food allergen, dust mites, pollen, mold or mold spores, weed pollen, tree pollen, grass pollen, fleas, pet dander, feathers or pet dander. Allergic skin reactions may include, for example, dermatitis, atopic dermatitis, contact dermatitis, eczema, atopic eczema, urticaria and psoriasis. Typically, it is a group of conditions that result in skin inflammation and symptoms including itching, redness of the skin and rash.

Allergic disorders may also include other allergic inflammatory conditions, such as eosinophilic esophagitis and other eosinophil-related gastrointestinal diseases. Eosinophilic esophagitis is an allergic inflammatory condition of the esophagus that involves eosinophils, a type of white blood cell. Symptoms include difficulty swallowing, food retention, vomiting and heartburn.

Butyrate, and therefore the compounds and compositions of the present invention, may prevent or reduce allergic reactions through one or more mechanisms.

The compounds and compositions of the present invention can modulate and/or stimulate Treg cell differentiation. Regulatory T cells (Treg) are very important for the induction of tolerance and thus can modulate and/or stimulate the induction of oral tolerance, leading to the prevention of food allergies or faster outgrowth of food allergies by regulating the type 2 allergic response and reducing production IgE. Many chronic inflammatory diseases, such as psoriasis, allergies and inflammatory bowel disease, are thought to develop as a result of a breakdown in tolerance. Preclinical data showed that direct oral administration of butyrate increased Treg cell differentiation (Tan, J., et al., 2016. Cell reports, 15(12), pp.2809-2824). Failure to induce Treg cell activity has been shown to result in impaired Th2 cell responses and the development of allergic disease. Abnormal T helper type 2 (Th2) cell responses can cause allergic inflammation. Type 2 innate lymphoid cells (ILC2s) are a critical source of Th2 cytokines, IL-5 and IL-13, which contribute to the exacerbation of acute asthma. Butyrate has been shown to be a very important regulator of ILC2 proliferation and function through its histone deacetylase (HDAC) inhibitory activity (Thio, C.L.P., et al. 2018. Journal of Allergy and Clinical Immunology, 142(6), pp.1867-1883.e12 ). In addition, antigen-specific stimulation of B cells in the presence of butyrate switched plasma cell differentiation towards the development of regulatory B cells (Breg) and IL-10 production (Shi, Y., et al., 2015. Scientific reports, 5, p. 17651).

The compounds and compositions of the present invention can increase the production and/or expression of one or more anti-inflammatory cytokines, and/or decrease the production and/or expression of one or more proinflammatory cytokines, and thereby reduce inflammation in tissues. In addition, butyrate has been shown to alter the production of anti-inflammatory cytokines, such as IL-10 (Shi, Y., et al., 2015. Scientific reports, 5, p.17651). Butyrate has also been shown to alter the production of a number of pro-inflammatory cytokines, for example, TNFalpha, IL-5, IL-13, IL-17 (Diakos, C., et al., 2006. Biochemical and biophysical research communications, 349(2), pp.863-868; Thio, C.L.P., et al. 2018. Journal of Allergy and Clinical Immunology, 142(6), pp.1867-1883.e12; and Singh, N., et al., 2014. Immunity, 40 (1), pp.128-139.).

The compounds and compositions of the present invention can modulate and/or reduce mast cell activity and all mast cell-mediated allergic symptoms such as diarrhea or urticaria. Butyrate has been shown to effectively inhibit mast cell activation and the production of inflammatory mediators in vivo (Wang, C.C., et al., 2018. Innate immunity, 24(1), pp.40-46).

The compounds and compositions of the present invention can attenuate the expression of GATA-3. GATA-3 is a transcription factor that is specifically expressed in T helper type 2 (Th2) cells and plays a critical role in the differentiation of Th2 cells from uncommitted CD4+ lymphocytes. Thus, the compound may reduce the formation of an allergic response. In addition, GATA-3 is required for the expression of the cytokine genes IL-4, IL-5 and IL-13, which mediate allergic inflammation (Barnes, P.J., 2008. Current molecular medicine, 8(5), pp.330-334). A high dose of sodium butyrate in vitro (1 mM) was sufficient to reduce Gata3 expression in Th2-polarized CD4+ T cells, followed by a switch to IFNγ (Kespohl, M., et al., 2017. Frontiers in immunology, 8, p. 1036). In other studies, 0.1 mM sodium butyrate was not sufficient to influence GATA-3 or FoxP3 expression in CD4+ T cells under Th2 polarization conditions (Furusawa, Y., et al., 2013. Nature, 504(7480), p.446).

Introduction

Preferably, the compounds and compositions described herein are administered enterally.

Enteral administration may be oral, gastric and/or rectal.

In one embodiment, administration is orally or via the stomach. In a preferred embodiment, administration is orally.

In general, administration of the combination or composition described herein to the gastrointestinal tract may, for example, be by the oral route or by other routes, for example, by tube.

The subject may be a mammal such as human, dog, cat, horse, goat, bull, sheep, pig, deer and primates. Preferably the subject is a human.

Organoleptic properties

The present invention provides compounds that are a source of butyrate having improved organoleptic properties. In particular, the compounds have an improved odor and/or taste compared to butyric acid, butyrate salts and/or tributyrin. In one embodiment, the compounds have an improved taste compared to tributyrin. In one embodiment, the compounds have an improved odor compared to butyrate salts (eg, sodium butyrate).

In one embodiment, the improved sensory properties are improved odor. In one embodiment, the improved sensory properties are improved taste. In one embodiment, the improved organoleptic properties are improved odor and improved taste. In one embodiment, the improved taste is a reduction in bitterness.

Examples

Example 1: Preparation of Butyrated Triglycerides (TAG)

Compositions containing butyrated TAGs were prepared by chemical transesterification between tributyrin and high oleic acid sunflower oil in the presence of a catalyst such as sodium methoxide. A molar excess of tributyrin relative to high oleic acid sunflower oil was used.

The three reactants, tributyrin, high oleic sunflower oil and catalyst, were mixed in a reactor under a nitrogen atmosphere and then heated with stirring at 80°C for 3 hours. After completion of the reaction, the product was washed several times with water, then dried under vacuum (25 mbar at 60°C for 2 hours). The resulting oil product was then subjected to a bleaching earth decolorization step and purified either by short-path distillation (130°C, 0.001–0.003 mbar) or deodorization (160°C, 2 mbar, 2 h) with steam injection.

The components of the resulting oil compositions, mainly triglycerides, are shown below in Table 1. These triglycerides are characterized by the content of three fatty acids. These fatty acids are characterized by lipid numbers: 4:0 for butyrate, 16:0 for palmitate, 18:0 for stearate, 18:1 for oleate and 18:2 for linoleate. The fatty acid listed in the middle is at the sn-2 position of the triglyceride. For example, 16:0 – 4:0 – 18:1 means two different triglycerides, each having a butyrate at the sn-2 position and either a palmitate at the sn-1 position and an oleate at the sn-3 position or an oleate at the sn- position 1 and palmitate at sn-3.

The TAG profile and regioisomers were analyzed using liquid chromatography coupled to a high-resolution mass spectrometer. The proportion of lipid classes was assessed using liquid chromatography coupled to an evaporative light scattering detector (ELSD).

Table 1. TAG regioisomeric profile [g/100 g]

Regioisomer TAG [g/100 g]

Composition

4 : 0 – 4 : 0 – 4 : 0 <0.4–4.7 4 : 0 – 16 : 0 – 4 : 0 0.8–1.0 4 : 0 – 18 : 2 – 4 : 0 4.0–6.3 4 : 0 – 4 : 0 – 18 : 1 3.0–6.1 4 : 0 – 18 : 1 – 4 : 0 16.2–27.0 4 : 0 – 18 : 0 – 4 : 0 0.8–1.3 4 : 0 – 22 : 0 – 4 : 0 ≤ 0.4 4 : 0 – 16 : 0 – 18 : 1 1.1–1.5 16 : 0 – 4 : 0 – 18 : 1 0.5–0.7 4 : 0 – 18 : 1 – 16 : 0 1.2–1.6 4 : 0 – 18 : 1 – 18 : 2 2.6–3.1 18 : 1 – 4 : 0 – 18 : 2 1.1–1.6 4 : 0 – 18 : 2 – 18 : 1 2.9–3.6 18:1 – 18:1 – 4:0 23.3–25.8 18 : 1 – 4 : 0 – 18 : 1 3.3–4.8 4 : 0 – 18 : 0 – 18 : 1 0.9–1.3 4 : 0 – 18 : 1 – 18 : 0 0.8–1.1 4 : 0 – 22 : 0 – 18 : 1 <0.4–0.5 18:1 – 18:1 – 16:0 0.6–1.4 18:1 – 18:1 – 18:2 1.3–1.5 18:1 – 18:2 – 18:1 0.5–0.7 18:1 – 18:1 – 18:1 6.1–10.7 18:1 – 18:1 – 18:0 0.5–0.8 In total 93.1–94.1

In composition samples, the two most common TAGs are 4:0 – 18:1 – 4:0 and 18:1 – 18:1 – 4:0, and together they amount to approximately 40 to 50 g/100 g.

Example 2. Aromatic properties of triglycerides containing a butyrate residue

The odor of a TAG solution containing a butyrate residue (consisting primarily of oleic and butyric fatty acids) was compared with a solution containing sodium butyrate.

Receiving a sample

Solutions of TAG containing a butyrate residue (see Example 1) or sodium butyrate were prepared and stored at 4°C before presentation to a panel of tasters. Each 250 mL solution contained 600 mg butyric acid (equivalent to one capsule of commercially available sodium butyrate supplement; concentration 2.4 mg/mL) and 1 w/v% BEBA Optipro 1 infant formula in acidified deionized water.

Samples were prepared the day before testing by placing 4 mL of each solution (TAG-butyrate solution; sodium butyrate solution) into Agilent vials.

Methodology

A test was carried out according to the “two out of five” scheme. In this test, the taster was provided with five samples. The taster was asked to identify two samples that differed from the other three. The order of sample presentation was randomized to avoid presentation order bias.

In addition to the two-out-of-five design, tasters were provided with a comment box to allow them to describe the nature of the perceived difference (e.g., odor intensity, odor quality).

results

Five samples were provided to tasters at a time. They were asked to open the cap, smell, and then close each bottle in a given order. The results are shown in Table 2.

table 2

Number of responses Number of correct answers Significance eleven 9 P < 0.0001***

P -value was calculated using the binomial test performed with Fizz software (Biosystemes, France).

Tasters who found the correct answers (the TAG containing the butyrate residue was different from sodium butyrate) noted that the sodium butyrate smelled like "cheese", while the TAG samples containing the butyrate residue had significantly less of this "cheese smell" and the odor was pretty neutral.

Example 3. Taste properties of triglycerides containing butyrate residue

An organoleptic comparison was made between a TAG solution containing a butyrate residue (see Example 1), consisting mainly of oleic and butyric acids, and a solution containing tributyrin.

Sample preparation

One scoop (4.6 g) of BEBA Optipro 1 infant formula was added to warm water (chilled boiled tap water as per instructions) to obtain a final volume of 150 ml (approximately 3 w/v solution). Each TAG form of butyrate was weighed separately to obtain 600 mg of butyrate, and infant formula was added to each solution to a final volume of 50 mL.

Solution A included TAG containing a butyrate moiety (see Example 1); and solution B contained tributyrin.

Methodology

The panel of tasters conducted a repeat tasting without coding information.

Samples were prepared immediately before preliminary bitterness assessment and each solution was shaken vigorously. Tasting cups labeled A and B were filled simultaneously with a small volume of the corresponding solution.

Two samples were presented to tasters simultaneously. They were asked to taste the solution by putting it in their mouth and spitting it out, and rate their perceived bitterness on a scale of 0–10; where 0 means no bitterness is felt, and 10 means it feels like the strongest imaginable bitterness.

results

The bitterness of solution A was rated by the panel of tasters as 4.33 ± 1.52, mean ± SD.

The bitterness of solution B was rated by the panel of tasters as 8.33 ± 1.52, mean ± SD.

These data indicate that the TAG formulation containing the butyrate moiety in the infant formula had a significantly less bitter taste compared to tributyrin in the infant formula.

Example 4. Taste properties of 1,3-dibutyryl-2-palmitoylglycerol

1,3-Dibutyryl-2-palmitoylglycerol (BPB) was synthesized as a single compound using the following synthesis scheme:

BPB was evaluated through a descriptive evaluation by a panel of tasters and found to have a neutral taste and odor.

Example 5: Preparation of Butyrated Triglycerides (TAG)

Compositions comprising triglycerides containing a butyrate moiety were prepared by the esterification reaction of monoolein (derived from sunflower oil) with butyric acid, which was added in molar excess (total 5 equivalents). These two reagents were mixed together in a flask and heated to reflux (the boiling point of butyric acid was 163.5°C). A condenser (colonne de Vigreux) was used to remove water. The reaction was monitored by thin layer chromatography (TLC) and stopped after all monoacylglycerol had been converted to triacylglycerol.

The components of the resulting oil compositions, mainly triglycerides, are shown below in Table 3. As in Example 1, triglycerides are characterized by the content of three fatty acids. These fatty acids are characterized by lipid numbers: 4:0 for butyrate, 16:0 for palmitate, 18:0 for stearate, 18:1 for oleate and 18:2 for linoleate. The fatty acid listed in the middle is at the sn-2 position of the triglyceride.

Table 3. Triglyceride profile [wt.%]

4: 0 – 4: 0 – 18: 1 and 4: 0 – 18: 1 – 4: 0 65.64 18:1 – 18:1 – 4:0 and 18:1 – 4:0 – 18:1 12.53 4: 0 – 4: 0 – 18: 2 and 4: 0 – 18: 2 – 4: 0 5.43 4: 0 – 4: 0 – 18: 0 and 4: 0 – 18: 0 – 4: 0 3.03 4 : 0 – 18 : 1 – 18 : 2 and isomers 2.98 4 : 0 – 16 : 0 – 18 : 1 and isomers 1.69 4 : 0 – 4 : 0 – 16 : 0 and 4 : 0 – 16 : 0 – 4 : 0 1.40 4 : 0 – 4 : 0 – 4 : 0 1.36 4 : 0 – 18 : 0 – 18 : 1 and isomers 0.99 4: 0 – 4: 0 – 22: 0 and 4: 0 – 22: 0 – 4: 0 0.82 18:1 – 18:1 – 18:1 0.63 4 : 0 – 22 : 0 – 18 : 1 and isomers 0.33 4 : 0 – 4 : 0 – 24 : 0 and 4 : 0 – 24 : 0 – 4 : 0 0.31 4: 0 – 4: 0 – 20: 0 and 4: 0 – 20: 0 – 4: 0 0.28 4 : 0 – 16 : 0 – 18 : 0 and isomers 0.25 18:0 – 18:0 – 16:0 0.22 4 : 0 – 16 : 0 – 18 : 2 and isomers 0.21 4: 0 – 4: 0 – 20: 1 and 4: 0 – 20: 1 – 4: 0 0.20 18:1 – 18:1 – 18:2 0.17 18:2 – 18:2 – 4:0 and 18:2 – 4:0 – 18:2 0.17 18:0 – 18:0 – 4:0 and 18:0 – 4:0 – 18:0 0.16 16:0 – 16:0 – 4:0 and 16:0 – 4:0 – 16:0 0.14 16:0 – 18:0 – 16:0 0.12 4: 0 – 4: 0 – 18: 3 and 4: 0 – 18: 3 – 4: 0 0.11 4: 0 – 4: 0 – 16: 1 and 4: 0 – 16: 1 – 4: 0 0.11

The most abundant triglyceride in the composition was determined to be 4:0 – 4:0 – 18:1.

The resulting oil product was then subjected to a bleaching earth decolorization step and purified either by short-path distillation (130°C, 0.001–0.003 mbar) or deodorization (160°C, 2 mbar, 2 h) with steam injection to remove residual reactants and intermediates such as butyric acid, MAG, and by-products such as DAG and tributyrin.

The resulting oil product was given a descriptive organoleptic evaluation and found to have better odor and taste than tributyrin and butyric acid.

Example 6: Addition of Butyrated Triglycerides (TAG) to Infant Formula and Reduction of Allergic Response in Pre-Allergic Mammals

Milk allergy was induced in female BALB/c mice at eight weeks of age by sensitization via subcutaneous application of the milk protein β-lactoglobulin (BLG) (“allergic group”). At this stage, all three “allergic groups” (allergic group 2, allergic group 2 and allergic group 3) were equally sensitized by cutaneous application of the milk protein β-lactoglobulin (BLG). The non-allergic negative control group received phosphate-buffered saline (PBS) only (“control group”). Three patches were applied, one week apart with no patch in between. Six days after the last skin patch application, mice were orally administered BLG following standard procedures known to those skilled in the art. The serum was processed for ELISA and BLG-specific immunoglobulin E (IgE) and BLG-specific immunoglobulin G1 (IgG1) were measured (Figs. 1 and 2, respectively). As can be seen in FIG. 1 and 2, mice in all three allergy groups 1, 2 and 3 showed an allergic response to oral challenge with BLG, indicating that the mice were sensitized to the milk protein β-lactoglobulin (BLG).

After induction of milk allergy, the “control group” and “allergy group 1” received a cow's milk allergy infant formula (in this example, a highly hydrolyzed infant formula commercially available under the brand name Althéra ®) diluted in drinking water, in over a three week period. “Allergy group 2” and “allergy group 3” received butyrate ad libitum in the presence of cow's milk allergy formula diluted in drinking water over a three-week period. Allergic group 2 received butyrate in the form of sodium butyrate, and allergic group 3 received butyrate in the form of a butyrated TAG composition according to Example 1 (referred to herein as “palatable butyrate”). Sodium butyrate or palatable butyrate was prepared at a final concentration of 600 μg of butyrate per milliliter of infant formula. Egg allergy was then induced in “allergic group 1,” “allergic group 2 sodium butyrate,” and “allergic group 3 palatable butyrate” by intraperitoneal administration of egg white ovalbumin (OVA) adjuvanted with aluminum hydroxide. The “control group” received only the adjuvant intraperitoneally. Two weeks after the last intraperitoneal administration, mice were orally challenged with OVA three times per week for a total of 12 challenges. All groups continued to receive infant formula +/- butyrate throughout the egg allergy development period. Mice were examined after each oral OVA challenge and assessed clinically. Allergy symptoms were defined as follows: 0 = normal stool; 1 = soft/sticky stool; 2 = unformed stool; 3 = loose stools/diarrhea; 4 = at least 2 episodes of liquid diarrhea; 5 = score 4 at the end of the study. These data were combined to produce a cumulative clinical score for each mouse in the study (Fig. 3). In fig. Figure 3 shows that “allergic group 2 treated with sodium butyrate” and “allergic group 3 treated with palatable butyrate” showed a reduction in allergic symptoms compared to “allergic group 1” not treated with butyrate. The serum was processed for ELISA, and OVA-specific IgE, OVA-specific IgG1, and mast cell protease 1 (MCPT-1) were measured (Figs. 4, 5, and 6, respectively). The similar concentrations of OVA-specific IgE and OVA-specific IgG1 observed in the three allergic groups indicate that the reduced allergic response after oral administration of OVA may not be due to decreased allergic sensitization to the allergen. In fig. Figure 6 shows a decrease in mast cell protease 1 levels in “allergic group 2 sodium butyrate” and “allergic group 3 palatable butyrate,” indicating modulation of the mast cell response in the butyrate-treated groups.

This example demonstrates that oral administration of butyrate to mammals with a pre-existing milk protein allergy reduces the allergic response to egg allergen.

All publications identified in the description above are incorporated herein by reference. Various modifications and variations of the described methods, cells, compositions and uses will be apparent to one skilled in the art without departing from the spirit and scope of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the claimed invention should not be unduly limited to such specific embodiments. In fact, it is intended that the following claims include various modifications to the described embodiments of the invention that will be apparent to those skilled in the art.

Claims (23)

1. The use of one or more compounds having the formula for the treatment or prevention of an allergic disorder, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently a long chain fatty acid containing from 16 to 20 carbon atoms. 2. Use of a composition containing one or more compounds having the formula for the treatment or prevention of an allergic disorder, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently a long chain fatty acid containing from 16 to 20 carbon atoms. 3. Use of the composition according to claim 2, wherein the composition contains a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4). 4. Use of a composition according to claim 2 or 3, wherein the compounds of formulas (1), (2), (3) and (4) constitute at least 50 wt.%, 60 wt.%, 70 wt.%, 80 wt. .%, 90 wt.%, 95 wt.% or 99 wt.% of the total triglyceride content in the composition. 5. Use of a composition according to claim 2, containing a compound of formula (4) as the main triglyceride containing a butyrate residue in the composition, wherein the compound of formula (4) constitutes at least 20 wt.%, at least 30 wt.%, at least 40 wt.%, at least 50 wt.%, at least 60 wt.%, at least 70 wt.%, at least 80 wt.% or at least 90 wt.% of the total content triglycerides containing a butyrate residue in the composition. 6. Use of the composition according to any one of paragraphs. 2-5, and compounds having formulas (1), (2), (3) and (4) constitute at least 50 wt.%, 60 wt.%, 70 wt.%, 80 wt.%, 90 wt.%, 95 wt.% or 99 wt.% of the total content of triglycerides containing butyrate residue in the composition. 7. Use of the composition according to any one of paragraphs. 2-6, wherein tributyrin constitutes less than 10 wt.% of the total triglyceride content in the composition, preferably less than 8 wt.%, more preferably less than 5 wt.% of the total triglyceride content in the composition. 8. Use of the composition according to any one of paragraphs. 2-7, and the composition additionally contains vitamin A and/or dietary fiber. 9. Use of the composition according to any one of paragraphs. 2-8, wherein the composition is a nutritional composition, wherein preferably the nutritional composition is a dietary supplement, infant formula, infant formula, beverage or pet care product. 10. Use of a compound or combination of compounds according to claim 1 or use of a composition according to any one of claims. 2-9, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and/or R ⁶ is an unsaturated fatty acid, preferably monounsaturated. 11. Use of a compound or combination of compounds according to claim 1 or 10 or use of a composition according to any one of claims. 2-10, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and/or R ⁶ are selected from the group consisting of oleic acid, palmitic acid or linoleic acid, preferably each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ represent oleic acid. 12. Use of a compound or combination of compounds according to any one of paragraphs. 1, 10 or 11 or use of a composition according to any one of paragraphs. 2-11, wherein the allergic disorder is selected from one or more of the group consisting of food allergy, food intolerance, respiratory allergy and skin allergy. 13. Use of a compound or combination of compounds according to any one of paragraphs. 1, 10-12 or use of a composition according to any one of paragraphs. 2-12, wherein the allergic disorder is selected from one or more of the group consisting of rhinitis, asthma, urticaria, hay fever, allergic conjunctivitis, dermatitis, atopic dermatitis, contact dermatitis, eczema, atopic eczema, urticaria, psoriasis, eosinophilic esophagitis and other eosinophilia-related gastrointestinal disease, allergic diarrhea, vomiting, abdominal pain or bloating. 14. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 10-13 or the use of a composition according to any one of paragraphs. 2-13, wherein the allergen in the allergic disorder is selected from one or more of food allergen, dust mite, pollen, mold or mildew spores, weed pollen, tree pollen, grass pollen, fleas, pet hair, feathers or pet dander. 15. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 10-14 or the use of a composition according to any one of paragraphs. 2-14, wherein the allergen in the allergic disorder is a food allergen, wherein preferably the food allergen is selected from nut, tree nut, peanut, fish, shellfish, crustacean, milk, egg, soy, gluten, grain, wheat, oat, barley, rye, celery, corn, lupine, sulfites, sesame, mustard, rice, poultry and meat. 16. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 10-15 or use of a composition according to any one of paragraphs. 2-15, wherein treatment or prevention of an allergic disorder includes reducing or preventing allergic symptoms in response to an allergen. 17. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 10-16 or the use of a composition according to any one of paragraphs. 2-16, wherein treatment or prevention of the allergic disorder includes increasing oral tolerance. 18. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 10-16 or the use of a composition according to any one of paragraphs. 2-16, wherein treatment or prevention of an allergic disorder includes reducing or preventing an allergic response to a subsequent allergen in an individual with an allergy. 19. Use of a compound or combination of compounds according to any one of paragraphs. 1 or 9-18 or the use of a composition according to any one of paragraphs. 2-18, and the compounds or combinations thereof have improved organoleptic properties compared to butyric acid, tributyrin and/or butyrate salts.

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