Dietary butyrate

Abstract

A compound of formula (1), (2), (3), (4) or a combination thereof for use in treating or preventing a cardiovascular disorder in a subject, wherein R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Independently a long chain fatty acid having 16 to 20 carbons.

Description

Technical Field

[0001] The present invention relates to a dietary source of butyrate esters with improved organoleptic properties for use in the treatment and / or prevention of cardiovascular disorders. Background Art

[0002] Cardiovascular disorders account for approximately 20% of deaths worldwide each year and remain the leading cause of death in both developed and developing countries (SK Wattanapitayakul et al., Pharmacology and Therapeutics, 89, 187-206 (2001)). Cardiovascular disorders are also the leading cause of disability later in life. Cardiovascular disorders affect the heart or blood vessels. The main cardiovascular disorder is atherosclerosis (also known as arteriosclerotic vascular disease), a condition in which plaque deposits of fatty substances (atheromas or atherosclerotic plaques) develop on the walls of medium-sized and large arteries, resulting in reduced or blocked blood flow. Atherosclerosis is a form of arteriosclerosis, which means hardening of the arteries. Hardening of the arteries interferes with the body's control of blood pressure, thereby increasing the risk of high blood pressure. The stiffness of the arteries prevents dilation that would otherwise restore blood pressure to normal. People with high blood pressure are at greater risk of stroke, heart attack, and kidney failure.

[0003] The main cause of atherosclerosis is unknown, but atherosclerosis is a syndrome that affects the arteries and is a chronic inflammatory response in the artery walls, largely caused by the accumulation of macrophages and inadequate removal of fats and cholesterol from macrophages by functional high-density lipoproteins (HDL), and is exacerbated by low-density lipoproteins (LDL, plasma lipoproteins that carry cholesterol and triglycerides). Atherosclerosis is often referred to as hardening or scaling of the arteries. This is caused by the formation of multiple plaques within the arteries.

[0004] In cases where atherosclerosis affects the arteries that supply blood to the heart (coronary artery disease), this can lead to chest pain (angina) or a heart attack in which areas of the heart muscle are destroyed (myocardial infarction). The reduction in the flow of oxygen-rich blood to the heart muscle can lead to heart failure, a disorder in which the heart cannot pump blood adequately, resulting in reduced blood flow, a backup (congestion) of blood in the veins and lungs, and other changes that can further weaken the heart. The inability of the coronary circulation to provide adequate circulation to the heart muscle and surrounding tissues is called coronary artery disease.

[0005] Atherosclerosis that affects the arteries of the brain can cause a stroke. A stroke occurs when a cerebral artery becomes blocked or ruptures, causing an area of ​​brain tissue to die (cerebral infarction).

[0006] Although the major cardiovascular disorders in terms of mortality are stroke and heart attack, cardiovascular disorders also encompass conditions such as aortic aneurysms and peripheral vascular disease, and lead to clinical conditions including renovascular disease, vascular dementia, and retinal disease. Preventing cardiovascular disorders involves not only reducing mortality, but also preventing disability and improving quality of life.

[0007] Initial treatment of cardiovascular disorders focuses on diet and lifestyle interventions, such as increasing physical activity, consuming a low-fat diet, and stopping smoking.

[0008] A large number of compounds for the treatment or prevention of cardiovascular disorders have been proposed. These include: salicylates (WO2009 / 006583), statins (JK Liao, International Journal of Cardiology, 86, 5-18 (2002)), angiotensin converting enzyme (ACE) inhibitors (US5684016), angiotensin II receptor blockers (EP1604664) and calcium channel blockers (EP0089167).

[0009] Unfortunately, currently available treatments are not always fully satisfactory, particularly in terms of efficacy and / or in terms of the side effects that may be associated with them.

[0010] It is known that many patients under statins are still in a state of increased CVD risk. Despite a significant reduction in LDL-C levels, hsCRP in these patients continues to rise. This defines the so-called "residual inflammatory risk" and has become a pharmacological target of interest. Evidence of the relevance of this target comes from the Canakinumab Anti-Inflammatory Thrombosis Outcome Study (CANTOS) trial. It is shown here that direct reduction of inflammation with IL-1β antagonists reduces cardiovascular event rates independently of LDL-C values ​​(PM Ridker, N Engl J Med. 2017 Sep 21; 377 (12): 1119-1131). However, IL-1β antagonist treatment has excessively high costs and important side effects.

[0011] It would therefore be highly desirable to have additional compositions useful for treating or preventing cardiovascular disorders without those disadvantages described in the prior art.

[0012] In the human large intestine, acetate, propionate and butyrate are the main short-chain fatty acids produced by the microbiota after fermentation of resistant starch or dietary fiber. Once produced, SCFAs are absorbed by intestinal enterocytes as an energy source, but a portion of these absorbed fatty acids reaches the systemic circulation and regulates the biological activity of multiple biological systems and / or organs. There is evidence that SCFAs can have anti-inflammatory effects and reduce the development of a variety of inflammatory diseases, including inflammatory bowel disease, colon cancer and diabetes (PAGill, Aliment Pharmacol Ther. 2018 Jul; 48 (1): 15-34).

[0013] However, the effects of SCFA on atherosclerosis and subsequent cardiovascular disease are still poorly understood. Arthrosis is a general term describing any hardening (and loss of elasticity) of medium or large arteries caused by atherosclerotic plaque formation in the arteries. Vascular researchers have previously documented the role of inflammation in atherosclerosis. During the initiation of atherosclerotic plaques, macrophages release proinflammatory cytokines that activate endothelial cells and recruit additional leukocytes. This results in leukocyte adhesion to endothelial cells, leukocyte transfer to the subendothelial cavity, and plaque progression and maturation. Internal plaques, inflammatory signals from activated leukocytes (especially IL1 and IL6) will eventually trigger plaque instability and rupture, as seen in myocardial infarction (PM Ridker, Circ Res. 2016 Jan 8; 118 (1): 145-56, PM Ridker, Circ Res. 2019 Feb; 124 (3): 437-450).

[0014] Recent clinical trial data have now confirmed that inflammation is a key driver of atherosclerosis, as previously proposed with epidemiological data. Circulating biomarkers of inflammation, including high-sensitivity C-reactive protein (hsCRP) and interleukin-6 (IL-6), are associated with an increased risk of cardiovascular events. This is independent of cholesterol and other traditional risk factors. In fact, randomized trials have shown that, in addition to its blood lipid-lowering effect (low-density lipoprotein cholesterol, also known as LDL-C), statin therapy also reduces hsCRP. Interestingly, the magnitude of the hsCRP reduction is proportional to the reduction in cardiovascular risk (PM Ridker, J Am Coll Cardiol. 2016 Feb 16; 67 (6): 712-723).

[0015] In vitro studies have demonstrated that endothelial inflammatory responses are also reduced by SCFA pretreatment after challenge. Endothelial cells challenged with LPS or TNF pretreated with SCFA secrete less interleukin-6, express less vascular cell adhesion protein 1 (VCAM1), and have reduced leukocyte adhesion. (M. Li, Front Pharmacol. 2018 May 23; 9: 533).

[0016] In other studies, butyrate has been found to significantly reduce the formation and activation of the Nrlp3 inflammasome induced by cholesterol crystals in cultured endothelial cells, while acetate or propionate did not significantly reduce it. Further in vivo studies have shown that butyrate has a protective effect on the arterial wall in a rodent model of carotid artery injury associated with poor Nrlp3 inflammasome complex formation and activation. (X. Yuan, Redox Biol. 2018 Jun; 16: 21-31).

[0017] These in vitro and in vivo studies support the anti-inflammatory effects of SCFAs and make butyrate a candidate for the prevention and / or treatment of atherosclerosis.

[0018] Common sources of butyrates are butyric acid and tributyrin, the latter being a triglyceride consisting of three ester functional groups with three butyrate moieties and a glycerol backbone. Butyric acid and tributyrin are generally regarded as safe (GRAS) (21 CFR582.60 and 21 CFR184.1903, respectively) food additives and are natural components of many dairy products. However, butyric acid is associated with negative organoleptic qualities, such as vomit-like, fecal-like and cheese-like aroma attributes. Tributyrin also has negative organoleptic qualities, particularly high bitterness. These unpleasant taste and odor attributes can make oral administration of compositions containing these compounds particularly difficult. The butyrate component from dairy products cannot be enriched, and therefore large amounts of dairy fat would need to be consumed, but this is not feasible for practical and nutritional reasons, especially because this would result in a large amount of undesirable calories derived from animal fat.

[0019] While butyrate may currently be beneficial in preventing and / or treating atherosclerosis, there are no methods to deliver butyrate in a form with acceptable organoleptic properties.

[0020] It would therefore be beneficial to provide a food-grade source of butyrate for the treatment or prevention of cardiovascular disorders having improved organoleptic properties compared to available solutions. Summary of the invention

[0021] The present invention provides compounds as a source of butyrates, the compounds having improved organoleptic properties for preventing or treating cardiovascular disease, the compounds having improved smell and / or taste relative to butyric acid, butyrate and / or tributyrin. The compounds can be used as a dietary source of butyric acid. The compounds can be used, for example, in nutritional compositions, dietary supplements, beverages and pet care products.

[0022] The compound and composition can be used to prevent or treat cardiovascular disorders, particularly atherosclerosis, by reducing the residual inflammatory risk. Cardiovascular disorders can be selected from atherosclerosis, coronary heart disease, myocardial infarction, angina pectoris, stroke and / or heart failure. As mentioned above, stroke and myocardial infarction (heart attack) are the main cardiovascular disorders in terms of mortality. Coronary heart disease and heart failure are also important causes of death and health failure. Atherosclerosis is the root cause of many other cardiovascular disorders. Many people endure regular outbreaks of high blood pressure and angina pectoris, but quality of life is often affected, and people have always paid attention to these diseases may be a prerequisite for stroke, heart failure or heart attack. Therefore, it will be advantageous to treat or prevent atherosclerosis, coronary heart disease, myocardial infarction, angina pectoris, stroke and / or heart failure.

[0023] According to a first aspect of the present invention, there is provided a compound having the formula

[0024]

[0025] or a combination thereof, for use in preventing or treating a cardiovascular disorder in an individual, wherein R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Independently a long chain fatty acid having 16 to 20 carbons.

[0026] According to another aspect of the present invention, a composition is provided, comprising a compound of formula (1), formula (2), formula (3) or formula (4), or a combination thereof, for preventing or treating cardiovascular disorders in an individual, wherein R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Independently a long chain fatty acid having 16 to 20 carbons.

[0027] In one embodiment, the composition comprises a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

[0028] The composition may comprise a compound having formula (1) and a compound having formula (2).

[0029] The composition may comprise a compound having formula (1) and a compound having formula (3).

[0030] The composition may comprise a compound having formula (1) and a compound having formula (4).

[0031] The composition may comprise a compound having formula (2) and a compound having formula (3).

[0032] The composition may comprise a compound having formula (2) and a compound having formula (4).

[0033] The composition may comprise a compound having formula (3) and a compound having formula (4).

[0034] The composition may comprise a compound having formula (1), a compound having formula (2), and a compound having formula (3).

[0035] The composition may comprise a compound having formula (1), a compound having formula (2), and a compound having formula (4).

[0036] The composition may comprise a compound having formula (1), a compound having formula (3), and a compound having formula (4).

[0037] The composition may comprise a compound having formula (2), a compound having formula (3), and a compound having formula (4).

[0038] The composition may include a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

[0039] In one embodiment, the compound of formula (1), the compound of formula (2), the compound of formula (3), and the compound of formula (4) comprise at least 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the total triglycerides in the composition.

[0040] In one embodiment, the compound of formula (1), the compound of formula (2), the compound of formula (3), and the compound of formula (4) comprise at least 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the total butyrate-containing triglycerides in the composition.

[0041] In one embodiment, tributyrin comprises less than 10 wt %, preferably less than 8 wt %, more preferably less than 5 wt % of the total triglycerides in the composition.

[0042] In one embodiment, the composition further comprises dietary fiber and / or probiotics.

[0043] The composition of the present invention may be in the form of a nutritional composition, such as a food, a beverage, a pet care product.

[0044] The composition of the present invention may be in the form of a dietary supplement.

[0045] In one embodiment, R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 It is an unsaturated fatty acid, preferably a monounsaturated one.

[0046] In one embodiment, R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 Selected from oleic acid, palmitic acid or linoleic acid, preferably R 1 , R 2 , R 3 , R 4 , R 5 and R 6 All are oleic acid.

[0047] The compounds and compositions of the present invention are useful for preventing or treating cardiovascular disorders.

[0048] The compounds and compositions of the present invention are useful for preventing the initiation and / or development of cardiovascular disorders, in particular atherosclerosis and atherosclerosis-related disorders, by reducing residual inflammatory risk.

[0049] In one embodiment, the compound or combination thereof has improved organoleptic properties relative to butyric acid, tributyrin and / or butyrate.

[0050] In one embodiment, the compound is administered to a mammal, preferably a human, a pet or a farm animal. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1Shown is the release of fatty acids from an emulsion containing 200 mg of a mixture of (A) tributyrin, (B) high oleic sunflower oil and (C) triacylglycerols (TAGs) containing a butyrate fraction according to the present invention, digested with (i) simulated intestinal fluid (SIF) or (ii) sequentially with gastric fluid (SGF) and simulated intestinal fluid (SIF).

[0052] Figure 2 The overall extent of lipid digestion of a mixture of tributyrin, high oleic sunflower oil and TAGs containing a butyrate fraction according to the invention after both SIF and SGF-SIF is shown. DETAILED DESCRIPTION

[0053] As used herein, the terms "comprising" and "consisting of" are synonymous with "including" or "containing" and are inclusive or open-ended and do not exclude additional unrecited members, elements or steps. The terms "comprising" and "consisting of" also include the term "consisting of".

[0054] Triglycerides

[0055] Triglycerides (also known as triacylglycerols) are triesters derived from glycerol and three fatty acids. Under hydrolysis conditions such as during digestion, triglycerides may be a source of fatty acids. For example, tributyrin may be a source of three moles of butyric acid per mole of tributyrin.

[0056] 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).

[0057] The term "fatty acid moiety" refers to the portion of a triglyceride produced from a fatty acid in an esterification reaction with glycerol. The triglyceride used in the present invention comprises at least one butyric acid moiety and at least one long chain fatty acid moiety.

[0058] Long chain fatty acids preferably used in the present invention are fatty acids having 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 compounds having the formula

[0059]

[0060] Or a combination thereof, for use in treating or preventing a cardiovascular disorder in a subject.

[0061] Other examples of triglycerides that can be used in the present invention include 1,3-dibutyryl-2-linoleoyl glycerol, 1,3-dibutyryl-2-stearoyl glycerol, 1-butyryl-2-oleoyl-3-palmitoyl glycerol, 1-palmitoyl-2-oleoyl-3-butyryl glycerol, 1-butyryl-2-oleoyl-3-linoleoyl glycerol, 1-linoleoyl-2-oleoyl-3-butyryl glycerol, 1-oleoyl-2-butyryl-3-linoleoyl glycerol, 1-linoleoyl-2-oleoyl-3-butyryl glycerol, 1-linoleoyl-2-butyryl-3-linoleoyl glycerol, 1-butyryl-2-oleoyl-3-oleoyl glycerol, 1-butyryl-2-oleoyl-3- Linoleoyl-3-oleoyl glycerol, 1-oleoyl-2-linoleoyl-3-butyryl glycerol, 1-butyryl-2-stearoyl-3-oleoyl glycerol, 1-oleoyl-2-stearoyl-3-butyryl glycerol, 1-butyryl-2-oleoyl-3-stearoyl glycerol, 1-stearoyl-2-oleoyl-3-butyryl glycerol, 1,2-dioleoyl-3-palmitoyl glycerol, 1-palmitoyl-2,3-dioleoyl glycerol, 1,2-dioleoyl-3-linoleoyl glycerol and 1-linoleoyl-2,3-dioleoyl glycerol.

[0062] The triglyceride of the present invention can be synthesized by, for example, esterification of long-chain fatty acids and butyric acid with glycerol.

[0063] The triglycerides of the present invention can be synthesized by, for example, transesterification between tributyrin and another triglyceride containing long-chain fatty acids. In one embodiment, high oleic sunflower oil is the source of long-chain fatty acids. This produces triglycerides containing primarily butyrate and oleate moieties. These compounds are dairy-free, cholesterol-free, and vegan. The fatty acids are released from the triglycerides by lipases naturally present in the gastrointestinal tract. Relative to butyrate, these compounds do not add additional mineral salts to the final formulation.

[0064] Alternative triglyceride synthesis methods can be routinely determined by those skilled in the art. By way of example, a method for obtaining 1,3-dibutyryl-2-palmitoylglycerol (BPB) is as follows:

[0065]

[0066] Triglycerides containing a single butyrate moiety may be used herein. Alternatively, mixtures of triglycerides containing different butyrate moieties may be used.

[0067] Composition

[0068] The compounds of the invention may be administered in the form of a composition. Accordingly, the invention provides a composition for treating or preventing a cardiovascular disorder in a subject, the composition comprising a triglyceride containing a butyrate moiety as described herein.

[0069] In one embodiment, the combination of a compound of formula (1) and a compound of formula (2) is present in a composition as defined herein.

[0070] In one embodiment, the compound of formula (1) is present in an amount of at least 10% by weight of the total triglycerides in the composition, and the compound of formula (2) is present in an amount of at least 10% by weight of the total triglycerides in the composition.

[0071] In one embodiment, the compound of formula (1) is present in an amount of at least 15% by weight of the total triglycerides in the composition, and the compound of formula (2) is present in an amount of at least 15% by weight of the total triglycerides in the composition.

[0072] In one embodiment, the compound of formula (1) is present in an amount of at least 20% by weight of the total triglycerides in the composition, and the compound of formula (2) is present in an amount of at least 20% by weight of the total triglycerides in the composition.

[0073] In one embodiment, the compound of formula (1) is present in an amount of at least 20% by weight of the total triglycerides in the composition, and the compound of formula (2) is present in an amount of at least 30% by weight of the total triglycerides in the composition.

[0074] In one embodiment, the compound of formula (1) comprises about 20% to about 40% by weight of the total triglycerides in the composition, and / or the compound of formula (2) comprises about 30% to about 40% by weight of the total triglycerides in the composition.

[0075] In one embodiment, the compound of formula (1) and the compound of formula (2) comprise at least 20%, 30%, 40%, 50%, 60% or 70% by weight of the total triglycerides in the composition, preferably from about 40% to about 80%, or from about 50% to about 75% by weight of the total triglycerides in the composition.

[0076] In one embodiment, the composition further comprises a compound of formula (3), preferably wherein the compound of formula (3) accounts for at least 2 weight %, 3 weight %, 4 weight % or 5 weight % of the total triglycerides in the composition, and / or the composition further comprises a compound of formula (4), preferably wherein the compound of formula (4) accounts for at least 1 weight %, 2 weight % or 3 weight % of the total triglycerides in the composition.

[0077] In one embodiment, the compound of formula (1) is present in an amount of at least 20 weight % of the total butyric acid-containing triglycerides in the composition, and the compound of formula (2) is present in an amount of at least 30 weight % of the total butyric acid-containing triglycerides in the composition.

[0078] In one embodiment, the compound of formula (1) comprises about 30% to about 50% by weight of the total butyric acid-containing triglycerides in the composition, and / or the compound of formula (2) comprises about 40% to about 60% by weight of the total butyric acid-containing triglycerides in the composition.

[0079] In one embodiment, the compound of formula (1) and the compound of formula (2) comprise at least 20%, 30%, 40%, 50%, 60%, 70% or 80% by weight of the total 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.

[0080] In one embodiment, the compound of formula (5) comprises at least 10% by weight of the total triglycerides in the composition and / or the compound of formula (6) comprises at least 10% by weight of the total triglycerides in the composition.

[0081] In one embodiment, the compound of formula (5) comprises at least 15% by weight of the total triglycerides in the composition and / or the compound of formula (6) comprises at least 15% by weight of the total triglycerides in the composition.

[0082] In one embodiment, the compound of formula (5) comprises at least 15% by weight of the total triglycerides in the composition and / or the compound of formula (6) comprises at least 20% by weight of the total triglycerides in the composition.

[0083] In one embodiment, the compound of formula (5) comprises at least 20% by weight of the total triglycerides in the composition and / or the compound of formula (6) comprises at least 20% by weight of the total triglycerides in the composition.

[0084] In one embodiment, the compound of formula (5) comprises from about 15% to about 30% by weight of the total triglycerides in the composition, and / or the compound of formula (6) comprises from about 20% to about 30% by weight of the total triglycerides in the composition.

[0085] In one embodiment, the compound of formula (5) and the compound of formula (6) comprise at least 20%, 30% or 40% by weight of the total triglycerides in the composition, preferably from about 30% to about 60% or from about 40% to about 50% by weight of the total triglycerides in the composition.

[0086] In one embodiment, the composition further comprises a compound having formula (7), preferably wherein the compound having formula (7) accounts for at least 2 weight % or 3 weight % of the total triglycerides in the composition, and / or the composition further comprises a compound having formula (8), preferably wherein the compound having formula (8) accounts for at least 2 weight % or 3 weight % of the total triglycerides in the composition.

[0087] In one embodiment, the compound of formula (5) comprises at least 10 weight percent of the total butyrate-containing triglycerides in the composition, and the compound of formula (6) comprises at least 10 weight percent of the total butyrate-containing triglycerides in the composition.

[0088] In one embodiment, the compound of formula (5) comprises at least 15% by weight of the total butyrate-containing triglycerides in the composition, and the compound of formula (6) comprises at least 15% by weight of the total butyrate-containing triglycerides in the composition.

[0089] In one embodiment, the compound of formula (5) comprises at least 15% by weight of the total butyrate-containing triglycerides in the composition, and the compound of formula (6) comprises at least 20% by weight of the total butyrate-containing triglycerides in the composition.

[0090] In one embodiment, the compound of formula (5) comprises at least 20 weight percent of the total butyrate-containing triglycerides in the composition, and the compound of formula (6) comprises at least 20 weight percent of the total butyrate-containing triglycerides in the composition.

[0091] In one embodiment, the compound of formula (5) comprises from about 15% to about 30% by weight of the total butyrate-containing triglycerides in the composition, and the compound of formula (6) comprises from about 20% to about 30% by weight of the total butyrate-containing triglycerides in the composition.

[0092] In one embodiment, the composition further comprises a compound of formula (7), preferably wherein the compound of formula (7) accounts for at least 2% or 3% by weight of the total butyrate-containing triglycerides in the composition, and / or the composition further comprises a compound of formula (8), preferably wherein the compound of formula (8) accounts for at least 2% or 3% by weight of the total butyrate-containing triglycerides in the composition.

[0093] In one embodiment, the composition of the present invention may also include 1,3-dibutyryl-2-linoleoyl glycerol, 1,3-dibutyryl-2-stearoyl glycerol, 1-butyryl-2-oleoyl-3-palmitoyl glycerol, 1-palmitoyl-2-oleoyl-3-butyryl glycerol, 1-butyryl-2-oleoyl-3-linoleoyl glycerol, 1-linoleoyl-2-oleoyl-3-butyryl glycerol, 1-oleoyl-2-butyryl-3-linoleoyl glycerol, 1-linoleoyl-2-oleoyl-3-butyryl glycerol, 1-oleoyl-2-butyryl-3-linoleoyl glycerol, 1-linoleoyl-2-butyryl-3-oleoyl glycerol, 1-butyryl-2- Oleyl-3-oleoyl glycerol, 1-oleoyl-2-linoleoyl-3-butyryl glycerol, 1-butyryl-2-stearoyl-3-oleoyl glycerol, 1-oleoyl-2-stearoyl-3-butyryl glycerol, 1-butyryl-2-oleoyl-3-stearoyl glycerol, 1-stearoyl-2-oleoyl-3-butyryl glycerol, 1,2-dioleoyl-3-palmitoyl glycerol, 1-palmitoyl-2,3-dioleoyl glycerol, 1,2-dioleoyl-3-linoleoyl glycerol and / or 1-linoleoyl-2,3-dioleoyl glycerol.

[0094] In one embodiment, tributyrin comprises less than 10 wt %, preferably less than 8 wt %, more preferably less than 5 wt % of the total butyrate-containing triglycerides in the composition.

[0095] The composition of the invention may be in the form of, for example, a solid (eg, powder), a liquid or a gel.

[0096] The composition of the present invention may be in the form of, for example, tablets, dragees, capsules, pellets, powders, granules, solutions, emulsions, suspensions, coated granules, spray-dried granules or pills.

[0097] The composition can be in the form of a pharmaceutical composition and can include 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 the Handbook of Pharmaceutical Excipients, 2nd Edition, (1994), Edited by A Wade and PJ Weller, edited by A Wade and PJ Weller. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical field and are described in, for example, Remington's Pharmaceutical Sciences, Mack Publishing Co., edited by A R Gennaro, 1985.

[0098] The pharmaceutical composition may include the following materials as carriers, excipients or diluents or in addition to carriers, excipients or diluents: any suitable binder, lubricant, suspending agent, coating agent and / or solubilizing agent. Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flowing lactose, beta-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, etc.

[0099] Preservatives, stabilizers, dyes and even flavoring agents may be provided in the composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents may also be used.

[0100] The composition may be a nutritional composition.

[0101] The expression "nutritional composition" refers to a composition that supplies nutrients to an individual. Such a nutritional composition is preferably orally administered and may comprise a lipid or fat source and a protein source. It may also contain a carbohydrate source. In one embodiment, the nutritional composition contains only a lipid or fat source. In other specific embodiments, the nutritional composition may comprise a lipid (or fat) source and a protein source, a carbohydrate source, or both.

[0102] In some embodiments, the nutritional composition according to the invention is an "enteral nutritional composition", i.e., it involves a food material to be administered in the gastrointestinal tract. Gastric introduction may involve the use of a tube passed through the oral / nasal passages or a tube in the abdomen directly to the stomach. This may be used in particular in a hospital or clinic.

[0103] In some embodiments, the composition is an oral nutritional supplement (ONS), a complete nutritional formula, a medicine, a medical product or a food product. In some embodiments, the composition is administered to an individual in the form of a beverage. The composition can be stored in a pouch in powder form and then suspended in a liquid such as water for use.

[0104] The composition according to the invention may be a dietary supplement.

[0105] The term "dietary supplement" can be used to supplement an individual's nutrition (it is usually used as such, but it can also be added to any kind of composition intended for ingestion). The supplement can be in tablet, capsule, lozenge or liquid form, for example. The supplement can also contain protective hydrophilic colloids (such as gums, proteins, modified starches), binders, film formers, encapsulating agents / materials, wall / shell materials, matrix compounds, coatings, emulsifiers, surfactants, solubilizers (oils, fats, waxes, lecithins, etc.), adsorbents, carriers, fillers, co-compounds, dispersants, wetting agents, processing aids (solvents), flow agents, taste masking agents, weighting agents, gelling agents and gel forming agents. Dietary supplements may also contain conventional pharmaceutical additives and adjuvants, excipients and diluents, including but not limited to: water, gelatin from any source, vegetable gums, lignin sulfonates, talc, sugars, starches, gum arabic, vegetable oils, polyalkylene glycols, flavors, preservatives, stabilizers, emulsifiers, buffers, lubricants, colorants, wetting agents, fillers, etc.

[0106] When the composition is a supplement, it may be provided in unit dosage form.

[0107] The composition according to the present invention may be a dairy product, a liquid beverage, a beverage powder, a dehydrated soup powder, a dietary supplement, a meal replacement, a nutritional bar, a cereal, a confectionery product or a dry pet food.

[0108] The composition may also include dietary fiber. "Dietary fiber" may include at least one non-digestible oligosaccharide (e.g., prebiotic). Prebiotics may be present in an amount of 0.3% to 10% by weight of the composition. Dietary fiber and / or prebiotics may promote the production of endogenous butyrate by intestinal microflora, thereby providing additional beneficial effects.

[0109] Prebiotics are generally non-digestible in the sense that they are not broken down and absorbed in the stomach or small intestine, and thus they can remain intact as they pass through the stomach and small intestine to the colon, where they are selectively fermented by beneficial bacteria. Examples of prebiotics include certain oligosaccharides, such as fructooligosaccharides (FOS), inulin, xylooligosaccharides (XOS), polydextrose, or any mixture thereof. In a specific embodiment, the prebiotic may be fructooligosaccharides and / or inulin. In a specific embodiment, the prebiotic is a combination of FOS and inulin, such as in the product sold by BENEO-Orafti under the trademark Fructo-oligosaccharides (formerly ) or sold by BENEO-Orafti under the trademark Inulin (formerly ). Another example is the combination of 70% short-chain oligofructose and 30% inulin, which is registered by Nestle under the trademark "Prebio 1". The nutritional composition of the present invention may also contain at least one milk oligosaccharide, which may be a BMO (bovine milk oligosaccharide) and / or an HMO (human milk oligosaccharide).

[0110] The composition of the present invention may also comprise at least one probiotic (or probiotic strain), such as a probiotic bacterial strain. Consumption of the probiotic strain may also promote the production of endogenous butyrate by the intestinal microflora, thereby providing additional beneficial effects.

[0111] The most commonly used probiotic microorganisms are mainly most bacteria and yeasts of the following genera: Lactobacillus spp., Streptococcus spp., Enterococcus spp., Bifidobacterium spp. and Saccharomyces pp.

[0112] In some embodiments, the probiotic is a probiotic bacterial strain. In some embodiments, it is a bifidobacterium and / or a lactobacillus.

[0113] Based on dry weight, the nutritional composition according to the invention may comprise 10e3 to 10e12 cfu of the probiotic strain, more preferably 10e7 to 10e12 cfu (such as 10e8 to 10e10 cfu) of the probiotic strain per g of composition.

[0114] In one embodiment, the probiotic is live. In another embodiment, the probiotic is non-replicating or inactivated. It can also be a probiotic part, such as a cell wall component or a product of probiotic metabolism. In some other embodiments, live probiotics and inactivated probiotics can be present simultaneously. The nutritional composition of the present invention may also include at least one bacteriophage (bacteriophage) or a mixture of bacteriophages, which are preferably directed against pathogenic Streptococci, Haemophilus, Moraxella and Staphylococcus.

[0115] The nutritional composition of the present invention typically comprises a protein source, a carbohydrate source and a lipid source. However, in some embodiments, particularly if the nutritional composition of the present invention is a supplement or fortifier, only lipid (or lipid source) may be present.

[0116] The nutritional composition according to the invention may contain a protein source. Protein sources based on, for example, whey, casein and mixtures thereof may be used, as well as soy-based protein sources. As far as whey protein is concerned, the protein source may be based on acid whey or sweet whey or a mixture thereof, and may contain α-lactalbumin and β-lactoglobulin in any desired proportion. In some embodiments, the protein source is mainly whey (i.e. more than 50% of the protein comes from whey protein, such as 60%> or 70%>). The protein may be intact or hydrolyzed, or a mixture of intact and hydrolyzed proteins. In some embodiments, the protein source may also be provided partially or completely in the form of added amino acids.

[0117] The term "intact" means that the main part of the protein is intact, i.e. the molecular structure has not changed, for example at least 80% of the protein has not changed, such as at least 85% of the protein has not changed, preferably at least 90% of the protein has not changed, even more preferably at least 95% of the protein has not changed, such as at least 98% of the protein has not changed. In a specific embodiment, 100% of the protein has not changed.

[0118] In the context of the present invention, the term "hydrolyzed" refers to a protein that has been hydrolyzed or broken down into its constituent amino acids.

[0119] The protein may be fully hydrolyzed or partially hydrolyzed. If hydrolyzed protein is desired, a hydrolysis process may be performed as desired and as known in the art. For example, a whey protein hydrolyzate may be prepared by enzymatic hydrolysis of the whey fraction in one or more steps. If the whey fraction used as a raw material is substantially lactose-free, it is found that the protein undergoes much less lysine blockage during the hydrolysis process. This makes it possible to reduce the degree of lysine blockage from about 15% by weight of total lysine to less than about 10% by weight of lysine; for example, about 7% by weight of lysine, which greatly improves the nutritional quality of the protein source.

[0120] In a specific embodiment, the protein of the composition is hydrolyzed, fully hydrolyzed or partially hydrolyzed. The degree of hydrolysis (DH) of the protein may be 2 to 20, 8 to 40, or 20 to 60, or 20 to 80, or greater than 10, 20, 40, 60, 80 or 90. For example, a nutritional composition containing a hydrolyzate with a degree of hydrolysis of less than about 15% is available from Nestle Company under the trademark Commercially available.

[0121] In some embodiments, the protein is substantially hydrolyzed.

[0122] At least 70%, 80%, 85%, 90%, 95% or 97% of the protein may be hydrolyzed. In a particular embodiment, 100% of the protein is hydrolyzed.

[0123] In a specific embodiment, protein is provided as amino acids.

[0124] In a specific embodiment, the protein of the composition is a plant-based protein.

[0125] The nutritional composition according to the invention may comprise a carbohydrate source. Any carbohydrate source commonly present in nutritional compositions may be used, such as lactose, sucrose, sucrose, maltodextrin, starch and mixtures thereof.

[0126] The nutritional composition of the present invention may also contain all vitamins and minerals that are considered to be necessary for daily diet and in a significant amount of nutrition. The minimum requirements of some vitamins and minerals have been determined. The examples of minerals, vitamins and other nutrients optionally present in the composition of the present 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 salt form. The presence and amount of specific minerals and other vitamins will vary according to the target population.

[0127] If necessary, the nutritional composition of the present invention may contain emulsifiers and stabilizers, such as soy, lecithin, citric acid monoglyceride and citric acid diglyceride, etc. The nutritional composition of the present invention may also contain other substances that may have beneficial effects, such as lactoferrin, osteopontin, TGFbeta, slgA, glutamine, nucleotides, nucleosides, etc.

[0128] The nutritional composition according to the present invention can be prepared in any suitable manner. For example, the composition can be prepared by blending the components together in appropriate portions, optionally with one or more carriers, and then mixing the dry blended mixture using a liquefier to form a liquid mixture. If the final product is to be a powder, the liquid mixture can then be homogenized, pasteurized, and optionally spray-dried. The composition can be homogenized before or after pasteurization.

[0129] The nutritional composition of the present invention can be administered to an individual such as a human, e.g., an elderly person, at a therapeutically effective dose. The therapeutically effective dose can be determined by a person skilled in the art and will depend on many factors known to those skilled in the art, such as the severity of the condition and the weight and general condition of the individual.

[0130] In one embodiment of the invention, the nutritional composition is administered to an individual in combination with an exercise or physical activity regimen.

[0131] The nutritional composition of the present invention can be formulated to be administered to an animal in the form of an animal treat (e.g., biscuits) or a dietary supplement. The composition can be a dry composition (e.g., kibble), a semi-wet composition, a wet composition, or any mixture thereof. In another embodiment, the nutritional composition is a dietary supplement such as gravy, drinking water, beverages, yogurt, powders, granules, pastes, suspensions, chews, small pieces, treats, snacks, pellets, pills, capsules, tablets, or any other suitable delivery form.

[0132] The nutritional composition may be administered to an individual in an amount sufficient to prevent or at least partially reduce the risk of developing a cardiovascular disorder, if the individual has not already developed the cardiovascular disorder. Such an amount is defined as a "prophylactically effective dose".

[0133] The nutritional composition is preferably administered daily or at least twice a week as a supplement to an individual's diet. In one embodiment, the composition is continuously administered to an individual for several days. For example, the composition can be administered daily to an individual for at least 30 consecutive days, 60 days, or 90 days. For another example, the composition can be administered to an individual over a longer period of time, such as 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years.

[0134] In a preferred embodiment, the nutritional composition is administered to the individual for a period of at least 3 months, such as 3 months to 1 year, and preferably at least 6 months.

[0135] The above application examples do not require continuous daily application without interruption. Instead, there may be some brief interruptions in the application, such as two to four days of interruption during the application. The ideal duration of application of the composition can be determined by those skilled in the art.

[0136] Cardiovascular disorders

[0137] The present invention provides compounds, compositions and methods for preventing and / or treating cardiovascular disorders by reducing the risk of residual inflammation.

[0138] Cardiovascular disorders affect the heart or blood vessels. The main cardiovascular disorder is atherosclerosis (also called arteriosclerotic vascular disease), a condition in which plaque deposits of fatty substances (atheromas or atherosclerotic plaques) develop on the walls of medium-sized and large arteries, resulting in reduced or blocked blood flow. Atherosclerosis is a form of arteriosclerosis, which means hardening of the arteries. Hardening of the arteries interferes with the body's control of blood pressure, increasing the risk of high blood pressure. The stiffness of the arteries prevents the dilation that would otherwise normalize blood pressure. People with high blood pressure are at greater risk of stroke, heart attack, and kidney failure.

[0139] Atherosclerosis is a syndrome that affects the arteries and is a chronic inflammatory response of the artery walls, caused largely by the accumulation of macrophages and inadequate removal of fats and cholesterol from macrophages by functional high-density lipoproteins (HDL), and exacerbated by low-density lipoproteins (LDL, a plasma lipoprotein that carries cholesterol and triglycerides). Atherosclerosis is often referred to as hardening or scaling of the arteries. This is caused by the formation of multiple plaques within the arteries.

[0140] In cases where atherosclerosis affects the arteries that supply blood to the heart (coronary artery disease), this can lead to chest pain (angina) or a heart attack in which areas of the heart muscle are destroyed (myocardial infarction). The reduction in the flow of oxygen-rich blood to the heart muscle can lead to heart failure, a disorder in which the heart cannot pump blood adequately, resulting in reduced blood flow, a backup (congestion) of blood in the veins and lungs, and other changes that can further weaken the heart. The inability of the coronary circulation to provide adequate circulation to the heart muscle and surrounding tissues is called coronary artery disease.

[0141] Atherosclerosis that affects the arteries of the brain can cause a stroke. A stroke occurs when a cerebral artery becomes blocked or ruptures, causing an area of ​​brain tissue to die (cerebral infarction).

[0142] Although the major cardiovascular disorders in terms of mortality are stroke and heart attack, cardiovascular disorders also encompass conditions such as aortic aneurysms and peripheral vascular disease, and lead to clinical conditions including renovascular disease, vascular dementia, and retinal disease. Preventing cardiovascular disorders involves not only reducing mortality, but also preventing disability and improving quality of life.

[0143] It will be appreciated that the compounds, compositions and methods of the invention may be beneficial in preventing and / or treating atherosclerosis and / or related disorders, in particular preventing the initiation or development of atherosclerosis.

[0144] Treatment

[0145] It will be appreciated that all references to treatment herein include curative, palliative and prophylactic treatment; however, in the context of the present invention, references to prevention are more often associated with prophylactic treatment. Treatment may also include arresting the progression of disease severity.

[0146] In one embodiment, the term "treatment" of any disease or disorder refers to improving the disease or disorder (i.e., slowing down or preventing or reducing the development of at least one clinical symptom in the disease or its clinical symptoms). In another embodiment, "treatment" refers to alleviating or improving at least one physical parameter, including those that the patient may not be able to recognize. In another embodiment, "treatment" refers to regulating the disease or disorder, whether physically (e.g., the stability of recognizable symptoms), physiologically (e.g., the stability of physical parameters) or both. In another embodiment, "treatment" refers to preventing or delaying the onset or development or progress of a disease or disorder. As used herein, if such an individual will benefit from such treatment biologically, medically or in quality of life, then the individual "needs treatment".

[0147] individual

[0148] The term "subject" refers to any animal, including humans and companion animals. Generally, the subject is a human or an avian, bovine, canine, equine, feline, caprine, murine, ovine, and porcine animal. The subject can be a horse or a companion animal, such as a cat or dog. Preferably, the subject is a human.

[0149] Treatment of mammals, particularly humans, is preferred. However, both human and veterinary treatments are within the scope of the present invention.

[0150] For veterinary subjects, canine, feline and equine subjects are preferred.

[0151] Application

[0152] Preferably, the compounds and compositions described herein are administered enterally.

[0153] Enteral administration may be oral, gastric and / or rectal administration.

[0154] In one embodiment, administration is oral or gastric administration. In a preferred embodiment, administration is oral administration.

[0155] Generally, administration of the combinations or compositions described herein may be, for example, by the oral route or another route into the gastrointestinal tract, such as by tube feeding.

[0156] The subject may be a mammal, such as a human, canine, feline, equine, caprine, bovine, ovine, porcine, cervidae, and primate. Preferably, the subject is a human.

[0157] Sensory properties

[0158] The present invention provides compounds as a source of butyrates having improved organoleptic properties. Specifically, the compounds have improved odor and / or taste relative to butyric acid, butyrate and / or tributyrin. In one embodiment, the compounds have improved taste relative to tributyrin. In one embodiment, the compounds have improved odor relative to butyrate (e.g., sodium butyrate).

[0159] In one embodiment, the improved organoleptic property is improved smell. In one embodiment, the improved organoleptic property is improved taste. In one embodiment, the improved organoleptic property is improved smell and improved taste. In one embodiment, the improved taste is reduced bitterness.

[0160] Example

[0161] Example 1 - Preparation of butyrated triglycerides (TAG)

[0162] The composition comprising butylated TAGs is produced by chemical transesterification between tributyrin and high oleic sunflower oil in the presence of a catalyst such as sodium methoxide. A molar excess of tributyrin is used compared to high oleic sunflower oil.

[0163] The three reagents, tributyrin, high oleic sunflower oil and catalyst, are mixed together in a reactor under a nitrogen atmosphere and then heated at 80°C for 3 hours with stirring. Once the reaction is complete, the product is washed several times with water and then dried under vacuum (25 mbar, 60°C, 2 hours). The resulting oil product is then subjected to a decolorization step under the action of bleaching earth and purified by short-path distillation (130°C, 0.001-0.003 mbar) or by deodorization by injection of steam water (160°C, 2 mbar, 2 hours).

[0164] The components of the resulting oil composition (mainly triglycerides) are shown in Table 1 below. These triglycerides are represented by the three fatty acids they contain. These fatty acids are represented by their lipid numbers: butyrate is 4:0, palmitate is 16:0, stearate is 18:0, oleate is 18:1, and linoleate is 18:2. The middle fatty acid is located at the sn-2 position in the triglyceride. For example, 16:0-4:0-18:1 represents two different triglycerides that have both sn-2 butyrate and sn-1 palmitate and sn-3 oleate or sn-1 oleate and sn-3 palmitate.

[0165] TAG distribution and regioisomers were analyzed by liquid chromatography coupled to a high-resolution mass spectrometer. The proportions of lipid classes were assessed by liquid chromatography coupled to an evaporative light scattering detector (ELSD).

[0166] Table 1: TAG regioisomer distribution [g / 100g]

[0167] TAG regioisomers [g / 100g]

[0168] Composition

[0169]

[0170]

[0171] In the composition samples, the two most abundant TAGs were 4:0-18:1-4:0 and 18:1-18:1-4:0, which together were about 40 g / 100 g to 50 g / 100 g.

[0172] Example 2 - Odor Properties of Triglycerides Containing Butyrate Moieties

[0173] The odor of a solution containing TAG with a butyrate moiety (composed primarily of oleic and butyric fatty acids) was compared to a solution containing sodium butyrate.

[0174] Sample preparation

[0175] Solutions containing TAGs with butyrate moieties (see Example 1) or containing sodium butyrate were prepared and stored at 4° C. before delivery to the sensory panel. Each 250 mL solution contained 600 mg butyric acid (equivalent to one commercially available sodium butyrate capsule as a supplement; 2.4 mg / mL concentration) and 1% w / v BEBA Optipro 1 infant formula in acidified deionized water.

[0176] The day before the assay, samples were prepared by placing 4 mL of each solution (butyric triglyceride solution; sodium butyrate solution) into Agilent vials.

[0177] method

[0178] A "two out of five test" was performed. In this test, five samples were presented to the panelists. The panelists were instructed to identify the two samples that were different from the other three samples. The order of presentation of the samples was randomized to avoid presentation order bias.

[0179] In addition to the 2 out of 5 test, panelists were provided with a comment box to allow them to comment on the nature of the perceived difference (eg, odor intensity, odor quality).

[0180] result

[0181] The five samples were presented to the panelists simultaneously. They were asked to uncap, sniff, and then cap each vial in the given order. The results are shown in Table 2.

[0182] Table 2

[0183] Number of responses Number of correct responses Significance 11 9 p<0.0001***

[0184] P values ​​were calculated using the binomial test performed by Fizz software (Biosystemes, France).

[0185] It was found that panelists who responded correctly (TAG containing a butyrate moiety was different from sodium butyrate) mentioned that sodium butyrate smelled like "cheese", while for the TAG samples containing a butyrate moiety, this "cheese" odor was significantly reduced and the smell was quite neutral.

[0186] Example 3 - Taste properties of triglycerides containing butyrate moieties

[0187] Solutions containing triglycerides with a butyrate moiety consisting mainly of oleic and butyric fatty acids (see Example 1) were subjected to a sensory benchmark test against solutions containing tributyrin.

[0188] Sample preparation :

[0189] One scoop (4.6 g) of BEBA Optipro 1 infant formula was added to warm water (cooled, boiled tap water as directed) to a final volume of 150 mL (approximately 3% w / v solution). Each butyrate in the form of a triglyceride was weighed separately to deliver 600 mg of butyrate, and infant formula was added to each solution to a final volume of 50 mL.

[0190] Solution A contained triglycerides having a butyrate moiety (see Example 1); and solution B contained tributyrin.

[0191] method

[0192] A group of panelists performed repeated blind tastings.

[0193] The samples were prepared immediately prior to the initial bitterness assessment and each solution was shaken vigorously. Tasting cups labeled A and B were simultaneously filled with a small amount of the respective solution.

[0194] The two samples were presented to the panelists simultaneously. They were asked to taste the solutions in small sips and rate the perceived bitterness on a scale of 0-10; where 0 is no perceived bitterness and 10 is similar to the maximum imaginable bitterness.

[0195] result

[0196] The panelists rated the bitterness of Solution A as 4.33 ± 1.52, mean ± SD.

[0197] The panelists rated the bitterness of Solution B as 8.33 ± 1.52, mean ± SD.

[0198] These data indicate that TAG compositions containing a butyrate moiety in infant formula are significantly less bitter than tributyrin in infant formula.

[0199] Example 4 - Taste Properties of 1,3-Dibutyryl-2-Palmitoyl Glycerol

[0200] 1,3-Dibutyryl-2-palmitoylglycerol (BPB) was synthesized as a single compound using the following synthesis:

[0201]

[0202] BPB was evaluated in a descriptive sensory panel evaluation and found to have a neutral taste and odor.

[0203] Example 5 - Digestion of triglycerides containing butyrate moieties

[0204] 5.1 Emulsion preparation

[0205] Prepare a mixture containing 0.3 wt% polyoxyethylene sorbitan monooleate ( 80) A stable 10 wt% oil-in-water emulsion. An ultrasonic probe homogenizer was then used to produce the emulsion.

[0206] 5.2 Particle size determination

[0207] The droplet size of each lipid emulsion was measured by laser light scattering using a Mastersizer 3000 equipped with a Hydro SM from Malvern Instruments (Malvern, Worcestershire, United Kingdom). The emulsion particle size is quoted as two values, the volume surface mean diameter D3,2 (D3,2 1 / 4Pnidi 3 / nidi 2) or volume length average diameter D4,3 (D4,3 1 / 4Pnidi 4 / nidi 3). The emulsion particle size results are the average of three measurements on two freshly prepared emulsions.

[0208] 5.3 In vitro digestion

[0209] The lipid emulsion (2mL) containing 200mg fat is subjected to gastrointestinal in vitro lipolysis. Digestion is carried out in a constant temperature glass container (37°C) in a pH-STAT setting controlled by TIM 856 double burette pH-STAT (Radiometer Analytical, France). For gastric digestion, the sample is incubated 90 minutes at 37°C and a pH of 5.5 with 8.5mL simulated gastric fluid (SGF), which is composed of 150mM NaCl, 450U / mL pepsin, 18U / mL rabbit gastric lipase. Digestion is initiated by adding the rabbit gastric lipase of 18 tributyrin U / ml (TBU) activity (measured when pH 5.4).

[0210] The intestinal digestion step is carried out in a pH stat, wherein the pH is kept constant at 6.8 by adding NaOH. A mixture of bile salts (bile salts prepared with tris buffer) and a calcium solution (20mM Ca, 176.5mM tris, 150mM NaCl) are added to the SGF sample mixture. The mixture is transferred to a pH-stat, wherein the pH is adjusted to about 6.78. The intestinal digestion step begins when the temperature reaches 37 ± 0.5 ° C. The pH is adjusted to pH 6.8, and after incubation for two minutes at this pH and temperature, a pancreatic enzyme solution is added. The final composition of the intestinal juice is 10mM CaCl2, 12mM mixed bile salts, 0.75mM phospholipids, 150mM NaCl and 4mM tris (hydroxymethyl) aminomethane buffer. The intestinal digestion step is carried out for 3 hours in a titration manager derived from Radiometer. During the intestinal phase of digestion, digestion kinetics were followed using pH-stat (TIM856, Radiometer) technology and expressed as titratable acids (rather than fatty acids) which can be calculated using the following formula:

[0211] TA=V NaOH ×0:05×1000

[0212] TA: total titratable acid released, mmol; V NaOH : Volume of NaOH used to titrate the released acid within 3 h, mL.

[0213] 5.6 Results

[0214] Since the digestion of dietary lipids involves lipases from both gastric and intestinal sources, lipid digestibility was assessed using two digestion models: i) simulated intestinal fluid (SIF) containing porcine pancreatic lipase (PPL), and ii) sequential digestion in simulated gastric fluid (SGF) containing rabbit gastric lipase (RGL) followed by simulated intestinal fluid (SIF) containing porcine pancreatic lipase (PPL). All lipids were digested using polyoxyethylene sorbitan monooleate ( 80) for emulsification and have similar particle size distribution and specific surface area ( Figure 2 ), which means that the digestion differences mainly come from the triglyceride molecular structure.

[0215] Figure 1 i AC shows the digestion of triglycerides of tributyrin (C4), high oleic sunflower oil (HOSFO, mainly C18:1) and the butyrate-containing fraction according to the invention, generated by chemical transesterification between tributyrin and high oleic sunflower oil (see Example 1) "C4-C18:1" and by porcine pancreatic lipase (from pancreatin) (SIF model) in the presence of mixed bile and calcium. The lipids generally exhibit the same lipolysis behavior, undergoing an initial rapid lipolysis phase within the first 15 minutes, which gradually slows down during the last 2.5 hours of simulated intestinal digestion. The C4 triglycerides exhibited 223±59 μmol.min -1 The initial maximum lipolysis rate of high oleic sunflower oil was 34.5±2.3μmol.min -1 Significantly lower (p<0.0001) than short-chain triglycerides. C4-C18:1 showed 153±47μmol.min -1 The initial hydrolysis rate of C4:1 was between that of C18:1. Overall, it can be seen that all triglycerides were rapidly and extensively digested in the presence of porcine pancreatic lipase.

[0216] Next, the triglyceride digestion was performed using the sequential SGF(RGL)SIF(PPL) model. The digestion in the SIF compartment is shown in Figure 1 ii AC. Measurements were not made in the gastric compartment due to limited ionization of the target fatty acids. C4 and C18:1 triglycerides generally released lower amounts of titratable acid during the 3 h digestion period compared to when digested with SIF alone. The effect was greatest with tributyrin, with an initial lipolysis rate of 223 ± 59 μmol.min when digested with SIF alone. -1 In comparison, the initial lipolysis rate during SGF-SIF digestion was 44.1 ± 8.8 μmol.min -1Significantly reduced (p<0.0001). The total amount of acid released after SGF-SIF digestion of tributyrin, 381±20 μmol, was almost 1 / 3 of the amount released after SIF digestion alone, 958±12.5 μmol. These results clearly indicate that there is extensive digestion of tributyrin in the gastric compartment of the model.

[0217] When exposed to SGF and SIF sequentially, the SIF lipolysis rate of triglycerides C4-C18:1 containing butyrate moieties was 124 ± 20 μmol.min -1 , compared with SIF alone (124 ± 20 μmol.min -1 ) showed a slight but insignificant decrease compared to the control group. The most interesting observation was the effect of secondary fatty acid chain length on the reduction in SIF lipolysis caused by RGL pre-exposure. Initially, tributyrin showed a 60.2% (147 ± 7.6 μmol) reduction in total fatty acid release during SIF lipolysis after pre-exposure to RGL in SGF. In contrast, C4-C18:1 interesterified triglycerides showed a 6.1% (45 ± 7.6 μmol) reduction.

[0218] Figure 2 The overall extent of lipid digestion after both SIF and SGF-SIF for three triglycerides using direct titration and reverse titration is shown in Figure 3. Because many fatty acids are only partially ionized at pH 6.8, direct titration only gives a partial picture of the extent of lipid digestion, and reverse titration to pH 11.5 or GC-FAME analysis is required to estimate the extent of complete digestion. The reverse titration results of the three triglycerides show that tributyrin and the triglyceride C4-C18:1 containing the butyrate moiety experienced 101.5 ± 0.9% and 101 ± 1.6% digestion, respectively, indicating that each molecule releases three fatty acids for complete digestion, while high oleic sunflower oil experienced 72.3 ± 2% digestion, indicating that each molecule releases two fatty acids for complete digestion.

[0219] In general, it can be seen that tributyrin undergoes extensive hydrolysis in the stomach, while high oleic sunflower oil triglycerides undergo very limited hydrolysis in the stomach. Surprisingly, it can be seen that triglycerides containing a butyrate moiety generated by transesterification of C4 with long-chain fatty acids (C4-C18:1) reduce the extent of gastric lipolysis of C4 fatty acids. Tributyrin undergoes approximately 60% lipolysis by gastric lipase, as indicated by a reduction in total fatty acid release during SIF lipolysis after pre-exposure to RGL in SGF. In contrast, triglycerides containing a butyrate moiety in C4-C18:1 showed only a 6.1% reduction in total fatty acid release in SGF-SIF. These results indicate that transesterification of C4 with long-chain fatty acids (C4-C18:1) regulates the release of butyric acid in the stomach, followed by release in the intestine after digestion, and that the design of structural lipids changes the timing (but not the extent) of delivery of short-chain fatty acids in the gastrointestinal tract.

[0220] All publications mentioned in the above description are incorporated herein by reference. Various modifications and variations of the methods, cells, compositions and uses disclosed in the present invention will be apparent to the skilled person without departing from the scope and spirit of the present invention. Although the present invention has been disclosed in conjunction with specific preferred embodiments, it should be understood that the present invention protected by the claims should not be unduly limited to such specific embodiments. In fact, various modifications to the mode disclosed for practicing the present invention that are apparent to the skilled person are intended to fall within the scope of the following claims.

Claims

1. Use of a composition comprising a compound of formula (1), a compound of formula (2), a compound of formula (3) and a compound of formula (4) in the preparation of a medicament for treating atherosclerosis in an individual, Where R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Independently a long chain fatty acid having 16 to 20 carbons.

2. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 50% by weight of the total triglycerides in the composition.

3. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 60% by weight of the total triglycerides in the composition.

4. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 70% by weight of the total triglycerides in the composition.

5. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 80% by weight of the total triglycerides in the composition.

6. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 90% by weight of the total triglycerides in the composition.

7. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 95% by weight of the total triglycerides in the composition.

8. Use of the composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 99% by weight of the total triglycerides in the composition.

9. The use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 50% by weight of the total butyrate-containing triglycerides in the composition.

10. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 60% by weight of the total butyrate-containing triglycerides in the composition.

11. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 70% by weight of the total butyrate-containing triglycerides in the composition.

12. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 80% by weight of the total butyrate-containing triglycerides in the composition.

13. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 90% by weight of the total butyrate-containing triglycerides in the composition.

14. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 95% by weight of the total butyrate-containing triglycerides in the composition.

15. Use of a composition according to claim 1, wherein the compound of formula (1), the compound of formula (2), the compound of formula (3) and the compound of formula (4) account for at least 99% by weight of the total butyrate-containing triglycerides in the composition.

16. Use of a composition according to any one of claims 1 to 15, wherein tributyrin represents less than 10% by weight of the total triglycerides in the composition.

17. Use of a composition according to any one of claims 1 to 15, wherein tributyrin represents less than 8% by weight of total triglycerides in the composition.

18. Use of a composition according to any one of claims 1 to 15, wherein tributyrin represents less than 5% by weight of the total triglycerides in the composition.

19. Use of a composition according to any one of claims 1 to 15, wherein the composition further comprises dietary fiber.

20. Use of a composition according to any one of claims 1 to 15, wherein the composition is a nutritional composition, a dietary supplement or a pet care product.

21. The use of a composition according to any one of claims 1 to 15, wherein R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 It is an unsaturated fatty acid.

22. The use of a composition according to any one of claims 1 to 15, wherein R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 It is a monounsaturated fatty acid.

23. The use of a composition according to any one of claims 1 to 15, wherein R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 Selected from oleic acid, palmitic acid or linoleic acid.

24. The use of a composition according to any one of claims 1 to 15, wherein R 1 , R 2 , R 3 , R 4 , R 5 and R 6 All are oleic acid.

25. Use of a composition according to any one of claims 1 to 15, wherein the composition has improved organoleptic properties relative to butyric acid, tributyrin and / or butyrate.

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