Dietary butyrates

Abstract

The present invention relates to dietary butyrates, in particular compounds having the formula (1), (2), (3), (4) wherein R1, R2, R3, R4, R5 and R6 are independently long chain fatty acids having 16 to 20 carbons, or combinations thereof, for use in the treatment or prevention of cardiovascular disorders in a subject. # imgabs0 #

Description

[0001] This application is a divisional application of the international application with application number PCT / EP2020 / 064068, titled "Dietary Butyrates", filed on May 20, 2020, which entered the Chinese national phase on November 18, 2021, with application number 202080036929.3. Technical Field

[0002] The present invention relates to dietary sources of butyrates with improved sensory properties for the treatment and / or prevention of cardiovascular disorders. Background Art

[0003] Cardiovascular disorders account for approximately 20% of the global annual death toll and remain the leading cause of death in both developed and developing countries (S.K. Wattanapitayakul et al., Pharmacology and Therapeutics, 89, 187 - 206 (2001)). Cardiovascular disorders are also the leading cause of disability in later life. Cardiovascular disorders affect the heart or blood vessels. The main cardiovascular disorder is atherosclerosis (also known as arteriosclerotic vascular disease), which is a condition in which deposits of fatty substances (atheromas or atherosclerotic plaques) develop on the walls of medium - sized and large arteries, leading to reduced or blocked blood flow. Atherosclerosis is a form of arteriosclerosis, which means hardening of the arteries. Arteriosclerosis interferes with the body's control of blood pressure, thus increasing the risk of hypertension. The stiffness of the arteries prevents the dilation that would otherwise normalize blood pressure. People with hypertension are at greater risk of stroke, heart attack, and kidney failure.

[0004] The main cause of atherosclerosis is still unknown, but atherosclerosis is a syndrome that affects arterial blood vessels and is also a chronic inflammatory response of the arterial wall, largely caused by the accumulation of macrophages and the insufficient removal of fat and cholesterol from macrophages by functional high - density lipoprotein (HDL), and is exacerbated by low - density lipoprotein (LDL, a plasma lipoprotein that carries cholesterol and triglycerides). Atherosclerosis is commonly referred to as the hardening or furring of arteries. This is due to the formation of multiple plaques within the arteries.

[0005] In the case 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 (myocardial infarction) in which areas of the heart muscle are damaged. The reduced flow of oxygen - rich blood to the myocardium can lead to heart failure, a disorder in which the heart pumps insufficiently, resulting in reduced blood flow, 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 sufficient circulation to the myocardium and surrounding tissues is called coronary heart disease.

[0006] Atherosclerosis affecting the cerebral arteries can lead to stroke. A stroke occurs when a cerebral artery is blocked or ruptured, resulting in the death of an area of brain tissue (cerebral infarction).

[0007] While the major cardiovascular disorders in terms of mortality are stroke and heart attack, cardiovascular disorders also encompass conditions such as aortic aneurysm and peripheral vascular disease, and result in clinical conditions including renal vascular disease, vascular dementia, and retinal disease. Prevention of cardiovascular disorders involves not only reducing mortality but also preventing disability and improving quality of life.

[0008] Initial treatment of cardiovascular disorders focuses on dietary and lifestyle interventions such as increasing exercise, consuming a low-fat diet, and quitting smoking.

[0009] Numerous compounds have been proposed for the treatment or prevention of cardiovascular disorders. These include: salicylates (WO2009 / 006583), statins (J.K. 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).

[0010] Regrettably, currently available treatments are not always entirely satisfactory, especially in terms of efficacy and / or in terms of the side effects that may be associated with them.

[0011] Many patients known to be on statins remain at increased risk of CVD. Despite a significant reduction in LDL-C levels, the 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 Outcomes Study (CANTOS) trial. It is shown here that direct reduction of inflammation with an IL-1β antagonist reduces the rate of cardiovascular events independently of LDL-C values (P.M. Ridker, N Engl J Med. 2017 Sep 21; 377(12):1119 - 1131). However, IL-1β antagonist treatment has excessive costs and significant side effects.

[0012] Accordingly, there is a great need for additional compositions that can be used for the treatment or prevention of cardiovascular disorders without the drawbacks described in the prior art.

[0013] In the human large intestine, acetate, propionate, and butyrate are the major short-chain fatty acids produced by the microbiota following fermentation of resistant starch or dietary fiber. Once produced, SCFAs are absorbed by enterocytes of the intestine as an energy source, but a portion of these absorbed fatty acids reach the systemic circulation and modulate the biological activity of multiple biological systems and / or organs. There is evidence that SCFAs can have anti-inflammatory effects and attenuate the development of multiple inflammatory diseases, including inflammatory bowel disease, colon cancer, and diabetes (P.A. Gill, Aliment Pharmacol Ther. 2018 Jul;48(1):15-34).

[0014] However, the effects of SCFAs on atherosclerosis and subsequent cardiovascular diseases remain poorly understood. Arteriosclerosis is a general term that describes any hardening (and loss of elasticity) of medium or large arteries due to the formation of atherosclerotic plaques in the arteries. Vascular researchers have previously documented the role of inflammation in atherosclerosis. During the initiation of atherosclerotic plaques, macrophages release pro-inflammatory cytokines that activate endothelial cells and recruit additional white blood cells. This leads to leukocyte adhesion to endothelial cells, leukocyte migration to the subendothelial space, and plaque progression and maturation. Inside the plaque, inflammatory signals from activated leukocytes (especially IL1 and IL6) will ultimately trigger plaque instability and rupture, as seen in myocardial infarction (P.M. Ridker, Circ Res. 2016 Jan 8;118(1):145-56, P.M. Ridker, Circ Res. 2019 Feb;124(3):437-450).

[0015] Recent clinical trial data have now confirmed that inflammation is a key driver of atherosclerosis, as previously proposed together 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, aka LDL-C), statin therapy also reduces hsCRP. Interestingly, the magnitude of hsCRP reduction is proportional to the reduction in cardiovascular risk (P.M. Ridker, J Am Coll Cardiol. 2016 Feb 16;67(6):712-723).

[0016] In vitro studies have demonstrated that, after an attack, the endothelial inflammatory response is also reduced by SCFA pretreatment. Endothelial cells challenged with LPS or TNF pretreated with SCFA secrete less interleukin-6, express less vascular cell adhesion protein 1 (VCAM1), and show reduced leukocyte adhesion. (M. Li, Front Pharmacol. 2018 May 23; 9:533).

[0017] In other studies, it has been found that in cultured endothelial cells, butyrate significantly reduces the formation and activation of the Nrlp3 inflammasome induced by cholesterol crystals, while acetate or propionate does 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 injury associated with the formation and activation of the adverse Nrlp3 inflammasome complex. (X. Yuan, Redox Biol. 2018 Jun; 16:21 - 31).

[0018] These in vitro and in vivo studies support the anti - inflammatory effects of SCFA and make butyrate a candidate for the prevention and / or treatment of atherosclerosis.

[0019] Common sources of butyrate are butyric acid and tributyrin, the latter being a triglyceride composed of three ester functional groups with three butyrate moieties and a glycerol backbone. Butyric acid and tributyrin are generally regarded as safe (GRAS) (21 CFR 582.60 and 21 CFR 184.1903, respectively) food additives and are natural components of many dairy products. However, butyric acid is associated with negative sensory qualities such as vomit - like, fecal - like, and cheesy aroma attributes. Tributyrin also has negative sensory qualities, especially high bitterness. These unpleasant taste and odor attributes can make oral administration of compositions containing these compounds particularly difficult. The butyrate component from dairy cannot be enriched and would therefore require consumption of large amounts of dairy fat, which is not practical and nutritionally unfeasible, especially as it would lead to a large amount of unwanted calories from animal fat.

[0020] Although currently butyrate may be beneficial for the prevention and / or treatment of atherosclerosis, there is no method to deliver butyrate in a form with acceptable sensory properties.

[0021] Therefore, it would be beneficial to provide a food - grade source of butyrate for the treatment or prevention of cardiovascular disorders with improved sensory properties compared to available solutions. SUMMARY OF THE INVENTION

[0022] The present invention provides compounds as a source of butyrate, which compounds have improved sensory properties for use in the prevention or treatment of cardiovascular diseases, and which compounds have improved odor 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.

[0023] The compounds and compositions 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, stroke, and / or heart failure. As described above, stroke and myocardial infarction (heart attack) are the leading cardiovascular disorders in terms of mortality. Coronary heart disease and heart failure are also important causes of death and health failure. Atherosclerosis is the underlying cause of many other cardiovascular disorders. Many people suffer from regular episodes of high blood pressure and angina, but the quality of life is often affected, and there is ongoing concern that these conditions can be a prerequisite for stroke, heart failure, or heart attack. Therefore, it would be advantageous to treat or prevent atherosclerosis, coronary heart disease, myocardial infarction, angina, stroke, and / or heart failure.

[0024] According to a first aspect of the present invention, there is provided a compound having the following formula

[0025]

[0026] or a combination thereof, which is used for preventing or treating cardiovascular disorders in an individual, wherein R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently long-chain fatty acids having 16 to 20 carbons.

[0027] According to another aspect of the present invention, there is provided a composition comprising a compound having formula (1), formula (2), formula (3), or formula (4), or a combination thereof, which is used for preventing or treating cardiovascular disorders in an individual, wherein R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently long-chain fatty acids having 16 to 20 carbons.

[0028] 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).

[0029] The composition may comprise a compound having formula (1) and a compound having formula (2).

[0030] The composition may comprise a compound having formula (1) and a compound having formula (3).

[0031] The composition may comprise a compound having formula (1) and a compound having formula (4).

[0032] The composition may comprise a compound having formula (2) and a compound having formula (3).

[0033] The composition may comprise a compound having formula (2) and a compound having formula (4).

[0034] The composition may comprise a compound having formula (3) and a compound having formula (4).

[0035] The composition may comprise a compound having formula (1), a compound having formula (2), and a compound having formula (3).

[0036] The composition may comprise a compound having formula (1), a compound having formula (2), and a compound having formula (4).

[0037] The composition may comprise a compound having formula (1), a compound having formula (3), and a compound having formula (4).

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

[0039] The composition may comprise a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

[0040] In one embodiment, the compound having formula (1), the compound having formula (2), the compound having formula (3), and the compound having formula (4) account for at least 50 wt%, 60 wt%, 70 wt%, 80 wt%, 90 wt%, 95 wt% or 99 wt% of the total triglycerides in the composition.

[0041] In one embodiment, the compound having formula (1), the compound having formula (2), the compound having formula (3), and the compound having formula (4) account for at least 50 wt%, 60 wt%, 70 wt%, 80 wt%, 90 wt%, 95 wt% or 99 wt% of the triglycerides of the total butyrate-containing moiety in the composition.

[0042] In one embodiment, tributyrin accounts for less than 10% by weight, preferably less than 8% by weight, more preferably less than 5% by weight of the total triglycerides in the composition.

[0043] In one embodiment, the composition further comprises dietary fiber and / or probiotics.

[0044] The composition of the present invention can be in the form of a nutritional composition, such as a food, a beverage, a pet care product.

[0045] The composition of the present invention can be in the form of a dietary supplement.

[0046] In one embodiment, R 1 、R 2 、R 3 、R 4 、R 5 and / or R 6 is an unsaturated fatty acid, preferably monounsaturated.

[0047] In one embodiment, R 1 、R 2 、R 3 、R 4 、R 5 and / or R 6 is selected from oleic acid, palmitic acid or linoleic acid, preferably R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are all oleic acid.

[0048] The compounds and compositions of the present invention can be used for preventing or treating cardiovascular disorders.

[0049] The compounds and compositions of the present invention can be used for preventing the initiation and / or development of cardiovascular disorders by reducing the residual inflammatory risk, specifically atherosclerosis and atherosclerosis-related disorders.

[0050] In one embodiment, the compounds or their combinations have improved sensory properties relative to butyric acid, tributyrin and / or butyrate.

[0051] In one embodiment, the compounds are provided to a mammal, preferably a human, a pet or a farm animal. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1Shows the release of fatty acids from an emulsion of a mixture containing 200 mg of (A) tributyrin, (B) high-oleic sunflower oil, and (C) a triacylglycerol (TAG) containing a butyrate moiety according to the present invention, digested with (i) simulated intestinal fluid (SIF) or (ii) sequentially with gastric fluid (SGF) and simulated intestinal fluid (SIF).

[0053] Figure 2 Shows the overall extent of lipid digestion of a mixture of tributyrin, high-oleic sunflower oil, and a TAG containing a butyrate moiety according to the present invention after both SIF and SGF - SIF. Detailed Description

[0054] As used herein, the terms "comprising" and "consisting of" are synonymous with "including" or "containing", and are inclusive of end values or open-ended, and do not exclude additional unlisted members, elements or steps. The terms "comprising" and "consisting of" also include the term "consisting of".

[0055] Triglyceride

[0056] Triglycerides (also known as triacylglycerols) are triesters derived from glycerol and three fatty acids. Under hydrolysis conditions such as during digestion, triglycerides can be a source of fatty acids. For example, tributyrin can be a source of three moles of butyric acid per mole of tributyrin.

[0057] Fatty acids are carboxylic acids with long tails (chains). Fatty acids can be unsaturated or saturated. Fatty acids not attached to other molecules are called free fatty acids (FFA).

[0058] The term "fatty acid moiety" refers to the part of a triglyceride produced by a fatty acid in an esterification reaction with glycerol. The triglycerides used in the present invention contain at least one butyric acid moiety and at least one long-chain fatty acid moiety.

[0059] The 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 following formula

[0060]

[0061] or combinations thereof, which are used for treating or preventing cardiovascular disorders in an individual.

[0062] 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-butyryl-3-oleoyl glycerol, 1-butyryl-2-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.

[0063] The triglycerides of the present invention can be synthesized, for example, by the esterification of long-chain fatty acids and butyric acid with glycerol.

[0064] The triglycerides of the present invention can be synthesized, for example, by the 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 mainly containing butyrate moieties and oleate moieties. These compounds are milk-free, cholesterol-free, and vegan. Fatty acids are released from the triglycerides due to lipases naturally present in the gastrointestinal tract. These compounds do not add additional mineral salts in the final formulation compared to butyrate.

[0065] Alternative methods for synthesizing triglycerides can be routinely determined by those skilled in the art. By way of example, the method for obtaining 1,3-dibutyryl-2-palmitoyl glycerol (BPB) is shown below:

[0066]

[0067] Triglycerides containing a single butyrate moiety can be used herein. Alternatively, a mixture of triglycerides containing different butyrate moieties can be used.

[0068] Composition

[0069] The compounds of the present invention can be administered in the form of a composition. Accordingly, the present invention provides a composition for treating or preventing cardiovascular disorders in an individual, the composition comprising a butyrate moiety-containing triglyceride as described herein.

[0070] In one embodiment, a combination of a compound having formula (1) and a compound having formula (2) is present in a composition as defined herein.

[0071] In one embodiment, the compound having formula (1) is present in an amount of at least 10% by weight of the total triglycerides in the composition, and the compound having formula (2) is present in an amount of at least 10% by weight of the total triglycerides in the composition.

[0072] In one embodiment, the compound having formula (1) is present in an amount of at least 15% by weight of the total triglycerides in the composition, and the compound having formula (2) is present in an amount of at least 15% by weight of the total triglycerides in the composition.

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

[0074] In one embodiment, the compound having formula (1) is present in an amount of at least 20% by weight of the total triglycerides in the composition, and the compound having formula (2) is present in an amount of at least 30% by weight of the total triglycerides in the composition.

[0075] In one embodiment, the compound having formula (1) accounts for about 20% to about 40% by weight of the total triglycerides in the composition, and / or the compound having formula (2) accounts for about 30% to about 40% by weight of the total triglycerides in the composition.

[0076] In one embodiment, the compound having formula (1) and the compound having formula (2) account for at least 20%, 30%, 40%, 50%, 60% or 70% by weight of the total triglycerides in the composition, preferably about 40% to about 80% by weight, or about 50% to about 75% by weight of the total triglycerides in the composition.

[0077] In one embodiment, the composition further comprises a compound having formula (3), preferably wherein the compound having formula (3) accounts for at least 2%, 3%, 4% or 5% by weight of the total triglycerides in the composition, and / or the composition further comprises a compound having formula (4), preferably wherein the compound having formula (4) accounts for at least 1%, 2% or 3% by weight of the total triglycerides in the composition.

[0078] In one embodiment, the compound having formula (1) is present in an amount of at least 20% by weight 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% by weight of the total butyric acid-containing triglycerides in the composition.

[0079] In one embodiment, the compound having the formula (1) accounts for about 30% to about 50% by weight of the total butyric acid-containing triglycerides in the composition, and / or the compound having the formula (2) accounts for about 40% to about 60% by weight of the total butyric acid-containing triglycerides in the composition.

[0080] In one embodiment, the compound having the formula (1) and the compound having the formula (2) account for at least 20%, 30%, 40%, 50%, 60%, 70% or 80% by weight of the total butyric acid-containing triglycerides in the composition, preferably about 60% to about 90% by weight of the total butyric acid-containing triglycerides in the composition.

[0081] In one embodiment, the compound having the formula (5) accounts for at least 10% by weight of the total triglycerides in the composition, and / or the compound having the formula (6) accounts for at least 10% by weight of the total triglycerides in the composition.

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

[0083] In one embodiment, the compound having the formula (5) accounts for at least 15% by weight of the total triglycerides in the composition, and / or the compound having the formula (6) accounts for at least 20% by weight of the total triglycerides in the composition.

[0084] In one embodiment, the compound having the formula (5) accounts for at least 20% by weight of the total triglycerides in the composition, and / or the compound having the formula (6) accounts for at least 20% by weight of the total triglycerides in the composition.

[0085] In one embodiment, the compound having the formula (5) accounts for about 15% to about 30% by weight of the total triglycerides in the composition, and / or the compound having the formula (6) accounts for about 20% to about 30% by weight of the total triglycerides in the composition.

[0086] In one embodiment, the compound having the formula (5) and the compound having the formula (6) account for at least 20%, 30% or 40% by weight of the total triglycerides in the composition, preferably about 30% to about 60% or about 40% to about 50% by weight of the total triglycerides in the composition.

[0087] In one embodiment, the composition further comprises a compound of formula (7), preferably wherein the compound of formula (7) comprises at least 2% or 3% by weight of the total triglycerides in the composition, and / or the composition further comprises a compound of formula (8), preferably wherein the compound of formula (8) comprises at least 2% or 3% by weight of the total triglycerides in the composition.

[0088] In one embodiment, the compound of formula (5) comprises at least 10% by weight of the triglycerides of the total butyrate-containing moiety in the composition, and the compound of formula (6) comprises at least 10% by weight of the triglycerides of the total butyrate-containing moiety in the composition.

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

[0090] In one embodiment, the compound of formula (5) comprises at least 15% by weight of the triglycerides of the total butyrate-containing moiety in the composition, and the compound of formula (6) comprises at least 20% by weight of the triglycerides of the total butyrate-containing moiety in the composition.

[0091] In one embodiment, the compound of formula (5) comprises at least 20% by weight of the triglycerides of the total butyrate-containing moiety in the composition, and the compound of formula (6) comprises at least 20% by weight of the triglycerides of the total butyrate-containing moiety in the composition.

[0092] In one embodiment, the compound of formula (5) comprises from about 15% to about 30% by weight of the triglycerides of the total butyrate-containing moiety in the composition, and the compound of formula (6) comprises from about 20% to about 30% by weight of the triglycerides of the total butyrate-containing moiety in the composition.

[0093] In one embodiment, the composition further comprises a compound of formula (7), preferably wherein the compound of formula (7) comprises at least 2% or 3% by weight of the triglycerides of the total butyrate-containing moiety in the composition, and / or the composition further comprises a compound of formula (8), preferably wherein the compound of formula (8) comprises at least 2% or 3% by weight of the triglycerides of the total butyrate-containing moiety in the composition.

[0094] In one embodiment, the composition of the present invention may further comprise 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-butyryl-3-oleoyl glycerol, 1-butyryl-2-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 / or 1-linoleoyl-2,3-dioleoyl glycerol.

[0095] In one embodiment, tributyrin accounts for less than 10% by weight, preferably less than 8% by weight, more preferably less than 5% by weight of the triglycerides of the total butyrate-containing portion in the composition.

[0096] The composition of the present invention can be, for example, in solid (such as powder), liquid or gel form.

[0097] The composition of the present invention can be, for example, in the form of tablets, dragees, capsules, soft gelatin capsules, powders, granules, solutions, emulsions, suspensions, coated granules, spray-dried granules or pellets.

[0098] The composition can be in the form of a pharmaceutical composition and may comprise one or more suitable pharmaceutically acceptable carriers, diluents and / or excipients. Examples of such suitable excipients of the compositions described herein can be found in "Handbook of Pharmaceutical Excipients", 2nd Edition, (1994), Edited by A Wade and PJ Weller. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in "Remington's Pharmaceutical Sciences", Mack Publishing Co., 1985, Edited by A.R. Gennaro.

[0099] The pharmaceutical composition may contain the following substances 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, β-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 and the like.

[0100] 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.

[0101] The composition may be a nutritional composition.

[0102] The expression "nutritional composition" refers to a composition that supplies nutrients to an individual. Such a nutritional composition is preferably oral and may contain 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 contain a lipid (or fat) source and a protein source, a carbohydrate source, or both.

[0103] In some specific embodiments, the nutritional composition according to the present invention is an "enteral nutritional composition", i.e., a foodstuff for administration in the gastrointestinal tract. Gastric introduction may involve the use of a tube passing through the mouth / nose passage or a tube leading directly to the stomach in the abdomen. This may be used especially in hospitals or clinics.

[0104] In some specific embodiments, the composition is an oral nutritional supplement (ONS), a complete nutritional formula, a medicine, a medical product or a food product. In some specific embodiments, the composition is administered to an individual in the form of a beverage. The composition may be stored in sachets in powder form and then suspended in a liquid such as water for consumption.

[0105] The composition according to the present invention may be a dietary supplement.

[0106] The term "dietary supplement" can be used to supplement an individual's nutrition (which is how it is commonly used, but it can also be incorporated into any type of composition intended for ingestion). Supplements can be, for example, in the form of tablets, capsules, lozenges or liquids. Supplements 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), glidants, taste masking agents, weight increasing agents, gelling agents and gel forming agents. Dietary supplements can also contain conventional pharmaceutical additives and adjuvants, excipients and diluents, including but not limited to: water, gelatin from any source, vegetable gums, lignosulfonates, talc, sugars, starches, gum arabic, vegetable oils, polyalkylene glycols, flavoring agents, preservatives, stabilizers, emulsifiers, buffers, lubricants, coloring agents, wetting agents, fillers, etc.

[0107] When the composition is a supplement, it can be provided in unit dosage form.

[0108] The composition according to the invention can be a dairy product, a liquid beverage, a beverage powder, a dehydrated soup powder, a dietary supplement, a meal replacement, a nutrition bar, a cereal, a confectionery product or a dry pet food.

[0109] The composition can also contain dietary fiber. "Dietary fiber" can contain at least one non-digestible oligosaccharide (such as a prebiotic). The prebiotic can be present in an amount of 0.3% to 10% by weight of the composition. The dietary fiber and / or prebiotic can promote the gut microbiota to produce endogenous butyrate, thus providing additional beneficial effects.

[0110] Prebiotics are generally non-digestible in the sense that they cannot be broken down and absorbed in the stomach or small intestine, and thus they can remain intact when passing through the stomach and small intestine to reach 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 can be fructooligosaccharides and / or inulin. In a specific embodiment, the prebiotic is a combination of FOS and inulin, such as in a product sold by BENEO-Orafti under the trademark Fructooligosaccharides (formerly ), or in a product sold by BENEO-Orafti under the trademark Inulin (formerly )Among the products for sale. Another example is a combination of 70% short-chain fructooligosaccharides and 30% inulin, which is registered by Nestle under the trademark "Prebio 1". The nutritional composition of the present invention may also comprise at least one milk oligosaccharide, which may be BMO (bovine milk oligosaccharide) and / or HMO (human milk oligosaccharide).

[0111] The composition of the present invention may also comprise at least one probiotic (or probiotic strain), such as a probiotic bacterial strain. Consuming a probiotic strain may also promote the gut microbiota to produce endogenous butyrate, thereby providing an additional beneficial effect.

[0112] 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 spp.

[0113] In some specific embodiments, the probiotic is a probiotic bacterial strain. In some specific embodiments, it is Bifidobacterium and / or Lactobacillus.

[0114] On a dry weight basis, the nutritional composition according to the present invention may comprise from 10e3 to 10e12 cfu of probiotic strain per g of the composition, more preferably from 10e7 to 10e12 cfu (such as 10e8 to 10e10 cfu) of probiotic strain.

[0115] In one embodiment, the probiotic is live. In another embodiment, the probiotic is non-replicating or inactivated. It may also be a probiotic moiety, such as a cell wall component or a product of probiotic metabolism. In some other embodiments, live probiotics and inactivated probiotics may be present simultaneously. The nutritional composition of the present invention may also comprise at least one bacteriophage (bacterial phage) or a mixture of bacteriophages, which are preferably directed against pathogenic Streptococci, Haemophilus, Moraxella and Staphylococci.

[0116] The nutritional composition of the present invention generally 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 a fortifier, only lipids (or a lipid source) may be present.

[0117] The nutritional composition according to the present invention may contain a protein source. Protein sources based on, for example, whey, casein, and mixtures thereof may be used, and protein sources based on soy may also be used. With respect to the whey proteins of interest, the protein source may be based on acid whey or sweet whey or a mixture thereof, and may contain any desired proportion of α-lactalbumin and β-lactoglobulin. In some embodiments, the protein source is whey-dominant (i.e., more than 50% of the protein is from whey proteins, such as >60% or >70%). The protein may be intact or hydrolyzed, or a mixture of intact protein and hydrolyzed protein. In some embodiments, the protein source may also be provided in part or in whole in the form of added amino acids.

[0118] The so-called term "intact" means that the major portion of the protein is intact, i.e., the molecular structure has not been altered, for example, at least 80% of the protein has not been altered, such as at least 85% of the protein has not been altered, preferably at least 90% of the protein has not been altered, even more preferably at least 95% of the protein has not been altered, such as at least 98% of the protein has not been altered. In a specific embodiment, 100% of the protein has not been altered.

[0119] In the context of the present invention, the term "hydrolyzed" means a protein that has been hydrolyzed or broken down into its constituent amino acids.

[0120] The protein may be completely hydrolyzed or partially hydrolyzed. If a hydrolyzed protein is desired, the hydrolysis process may be carried out as needed and as known in the art. For example, whey protein hydrolysates may be prepared by enzymatic hydrolysis of whey fractions in one or more steps. If the whey fraction used as a raw material is substantially lactose-free, it has been found that the protein undergoes much less lysine blockage during the hydrolysis process. This enables the degree of lysine blockage to be reduced from about 15 wt% of the total lysine to less than about 10 wt% of lysine; for example, about 7 wt% of lysine, which greatly improves the nutritional quality of the protein source.

[0121] 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 less than about 15% may be commercially available from Nestle Company under the trademark Commercially available.

[0122] In some embodiments, the protein is fully hydrolyzed.

[0123] At least 70%, 80%, 85%, 90%, 95% or 97% of the protein can be hydrolyzed. In a specific embodiment, 100% of the protein is hydrolyzed.

[0124] In a specific embodiment, the protein is provided as amino acids.

[0125] In a specific embodiment, the protein of the composition is a plant-based protein.

[0126] The nutritional composition according to the invention may comprise a carbohydrate source. Any carbohydrate source commonly present in nutritional compositions can be used, such as lactose, sucrose, cane sugar, maltodextrin, starch, and mixtures thereof.

[0127] The nutritional composition of the invention may also contain all the vitamins and minerals considered essential for the daily diet and in nutritionally significant amounts. The minimum requirements for certain vitamins and minerals have been determined. Examples of minerals, vitamins, and other nutrients 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 target group.

[0128] If necessary, the nutritional composition of the invention may contain emulsifiers and stabilizers, such as soy, lecithin, glycerol monostearate citrate, and glycerol distearate citrate, etc. The nutritional composition of the invention may also contain other substances that may have beneficial effects, such as lactoferrin, osteopontin, TGFbeta, slgA, glutamine, nucleotides, nucleosides, etc.

[0129] The nutritional composition according to the invention can be prepared in any suitable manner. For example, the composition can be prepared by blending the components together in appropriate proportions, optionally blending with one or more carriers, and then using a liquefier to mix the dry blend 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.

[0130] The nutritional composition of the invention can be administered to an individual such as a human, for example an elderly person, in 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 a person skilled in the art, such as the severity of the condition and the weight and general condition of the individual.

[0131] In one embodiment of the present invention, a nutritional composition is administered to an individual in combination with an exercise or physical activity program.

[0132] 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., a biscuit) or a dietary supplement. The composition can be a dry composition (e.g., kibble), a semi-moist composition, a wet composition, or any mixture thereof. In another embodiment, the nutritional composition is a dietary supplement, such as gravy, drinking water, a beverage, yogurt, a powder, a granule, a paste, a suspension, a chew, a nugget, a treat, a snack, a bolus, a pill, a capsule, a tablet, or any other suitable delivery form.

[0133] The nutritional composition can be administered to an individual in an amount sufficient to prevent or at least partially reduce the risk of developing a cardiovascular disorder in a situation where the individual has not yet developed a cardiovascular disorder. Such an amount is defined as a "preventive effective dose".

[0134] The nutritional composition is preferably administered daily or at least twice a week as a supplement to the individual's diet. In one embodiment, the composition is administered continuously to the individual for several days. For example, the composition can be administered daily to the individual for at least 30, 60, or 90 consecutive days. Alternatively, the composition can be administered to the individual for a longer period of time, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years.

[0135] In a preferred embodiment, the nutritional composition is administered to the individual for a period of at least 3 months, such as from 3 months to 1 year, and preferably for at least 6 months.

[0136] The above examples of administration do not require uninterrupted continuous daily administration. Instead, there can be some short breaks in the administration, such as a break of two to four days during the administration. The desired duration of administering the composition can be determined by those skilled in the art.

[0137] Cardiovascular disorder

[0138] The present invention provides compounds, compositions, and methods for preventing and / or treating cardiovascular disorders by reducing the risk of residual inflammation.

[0139] Cardiovascular disorders affect the heart or blood vessels. The major cardiovascular disorder is atherosclerosis (also known as arteriosclerotic vascular disease), a condition in which fatty plaques (atheromas or atherosclerotic plaques) develop on the walls of medium-sized and large arteries, leading to reduced or blocked blood flow. Atherosclerosis is a form of arteriosclerosis, which means hardening of the arteries. Arteriosclerosis interferes with the body's control of blood pressure, increasing the risk of hypertension. The stiffness of the arteries prevents the dilation that would otherwise return blood pressure to normal. People with hypertension are at greater risk of stroke, heart attack, and kidney failure.

[0140] Atherosclerosis is a syndrome that affects arterial blood vessels and is also a chronic inflammatory response of the arterial wall, largely caused by the accumulation of macrophages and the insufficient removal of fat and cholesterol from macrophages by functional high-density lipoprotein (HDL), and is exacerbated by low-density lipoprotein (LDL, a plasma lipoprotein that carries cholesterol and triglycerides). Atherosclerosis is commonly referred to as hardening or furring of the arteries. This is due to the formation of multiple plaques within the arteries.

[0141] In the case 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 (myocardial infarction) in which an area of the heart muscle is damaged. Reduced flow of oxygen-rich blood to the myocardium can lead to heart failure, a disorder in which the heart pumps insufficiently, resulting in reduced blood flow, backup (congestion) of blood in the veins and lungs, and other changes that can further weaken the heart. Inability of the coronary circulation to provide adequate circulation to the myocardium and surrounding tissues is called coronary heart disease.

[0142] Atherosclerosis affecting the arteries of the brain can lead to stroke. A stroke occurs when a brain artery is blocked or ruptured, resulting in death of an area of brain tissue (cerebral infarction).

[0143] Although the major cardiovascular disorders in terms of mortality are stroke and heart attack, cardiovascular disorders also encompass conditions such as aortic aneurysm and peripheral vascular disease, and result in clinical conditions including renal vascular disease, vascular dementia, and retinal disease. Prevention of cardiovascular disorders involves not only reducing mortality but also preventing disability and improving quality of life.

[0144] It should be understood that the compounds, compositions, and methods of the present invention may be beneficial for the prevention and / or treatment of atherosclerosis and / or related conditions, specifically for preventing the initiation or development of atherosclerosis.

[0145] Treatment method

[0146] It should be recognized 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 can also include arresting the progression of disease severity.

[0147] In one embodiment, "treatment" of any disease or disorder refers to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of at least one clinical symptom of the disease or its clinical symptoms). In another embodiment, "treatment" refers to alleviating or ameliorating at least one physical parameter, including those that may not be recognized by the patient. In another embodiment, "treatment" refers to modulating the disease or disorder, whether physically (e.g., stabilization of recognizable symptoms), physiologically (e.g., stabilization of physical parameters), or both. In another embodiment, "treatment" refers to preventing or delaying the onset or development or progression of a disease or disorder. As used herein, an individual "in need of treatment" if such individual would benefit biologically, medically, or in terms of quality of life from such treatment.

[0148] Individual

[0149] The term "individual" refers to any animal, including humans and companion animals. Generally speaking, an individual is a human or an avian, bovine, canine, equine, feline, caprine, murine, ovine, and porcine animal. An individual can be a horse or a companion animal, such as a cat or a dog. Preferably, the individual is human.

[0150] Treatment of mammals, particularly humans, is preferred. However, both human and veterinary treatments are within the scope of the present invention.

[0151] For veterinary individuals, dogs, cats, and equine individuals are preferred.

[0152] Administration

[0153] Preferably, the compounds and compositions described herein are administered enterally.

[0154] Enteral administration can be oral, gastric, and / or rectal administration.

[0155] In one embodiment, the administration is oral or gastric administration. In a preferred embodiment, the administration is oral administration.

[0156] Generally speaking, the administration of the combinations or compositions described herein can enter the gastrointestinal tract, for example, through the oral route or another route, such as by gavage administration.

[0157] The individual can be a mammal such as a human, a canine, a feline, a equine, a goat, a bovine, a sheep, a pig, a cervid, and a primate. Preferably, the individual is a human.

[0158] Sensory property

[0159] The present invention provides a compound as a source of butyrate esters, which compound has improved sensory properties. Specifically, the compound has improved odor and / or taste relative to butyric acid, butyrates, and / or tributyrin. In one embodiment, the compound has improved taste relative to tributyrin. In one embodiment, the compound has improved odor relative to butyrates (such as sodium butyrate).

[0160] In one embodiment, the improved sensory property is improved odor. In one embodiment, the improved sensory property is improved taste. In one embodiment, the improved sensory property is improved odor and improved taste. In one embodiment, the improved taste is reduced bitterness.

[0161] Example

[0162] Example 1 - Preparation of butyrylated triglyceride (TAG)

[0163] A composition comprising butyrylated TAG 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.

[0164] The three reagents, tributyrin, high oleic sunflower oil, and the catalyst, are mixed together into a reactor under a nitrogen atmosphere and then heated at 80 °C with stirring for 3 hours. Once the reaction is complete, the product is washed several times with water and then dried under vacuum (25 mbar, 60 °C, 2 hours). Then a decolorization step is carried out on the resulting oil product under the action of bleaching earth and purification is carried out by short path distillation (130 °C, 0.001 - 0.003 mbar) or by deodorization by injecting steam water (160 °C, 2 mbar, 2 hours).

[0165] The composition of the resulting oil (mainly triglycerides) is 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 in the sn-2 position in the triglyceride. For example, 16:0-4:0-18:1 represents two different triglycerides that have both a butyrate in the sn-2 position and a palmitate in the sn-1 position and an oleate in the sn-3 position or an oleate in the sn-1 position and a palmitate in the sn-3 position.

[0166] TAG distribution and regioisomers were analyzed by liquid chromatography coupled with high-resolution mass spectrometry. The proportion of lipid classes was evaluated by liquid chromatography coupled with evaporative light scattering detector (ELSD).

[0167] Table 1: TAG regioisomer distribution [g / 100g]

[0168] TAG regioisomers [g / 100g]

[0169] Composition

[0170]

[0171]

[0172] In the composition sample, 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 / 100g to 50 g / 100g.

[0173] Example 2 - Odor properties of triglycerides containing butyrate moieties

[0174] A solution containing TAGs with butyrate moieties (mainly composed of oleic acid and butyric fatty acids) was compared in terms of odor with a solution containing sodium butyrate.

[0175] Sample preparation

[0176] Solutions containing TAGs with butyrate moieties (see Example 1) or containing sodium butyrate were prepared and stored at 4 °C before being delivered to the sensory panel. Each 250 mL solution contained 600 mg butyric acid (equivalent to a commercially available sodium butyrate capsule as a supplement; 2.4 mg / mL concentration) and an acidified deionized aqueous solution of 1% w / v BEBA Optipro 1 infant formula.

[0177] One day before the test, samples were prepared by placing 4 mL of each solution (triglyceride butyrate solution; sodium butyrate solution) into an Agilent sample vial.

[0178] Method

[0179] A "two-out-of-five test" was conducted. In this test, panelists were presented with five samples. Panelists were instructed to identify the two samples that were different from the other three samples. The presentation order of the samples was randomized to avoid presentation order bias.

[0180] In addition to the two-out-of-five test, comment boxes were provided to panelists to allow them to comment on the nature of the perceived differences (e.g., odor intensity, odor quality).

[0181] Result

[0182] Five samples were presented to the panelists simultaneously. They were asked to open the lids, sniff, and then close each vial in the given order. The results are shown in Table 2.

[0183] Table 2

[0184] Number of responses Number of correct responses Significance 11 9 p<0.0001***

[0185] The P-value was calculated using the binomial test performed by Fizz software (Biosystemes, France).

[0186] Panelists who detected the correct response (TAG with butyrate moiety different from sodium butyrate) mentioned that sodium butyrate smelled like "cheese", while for the TAG sample with butyrate moiety, this "cheese" odor was significantly reduced and the odor was rather neutral.

[0187] Example 3 - Taste properties of triglycerides containing butyrate moieties

[0188] A solution containing a triglyceride mainly composed of oleic and butyric fatty acids with a butyrate moiety (see Example 1) was subjected to a sensory benchmark test against a solution containing tributyrin.

[0189] Sample preparation :

[0190] One scoop (4.6 g) of BEBA Optipro 1 infant formula was added to warm water (tap water cooled and boiled according to the instructions) to a final volume of 150 mL (approx. 3% w / v solution). Each triglyceride form of butyrate was weighed separately to deliver 600 mg of butyrate, and the infant formula was added to each solution to a final volume of 50 mL.

[0191] Solution A contained a triglyceride with a butyrate moiety (see Example 1); and solution B contained tributyrin.

[0192] Method

[0193] A group of panelists performed repeated blind tastings.

[0194] Samples were prepared just prior to the initial bitterness assessment and each solution was shaken vigorously. Tasting cups labeled A and B were filled simultaneously with a small amount of the respective solution.

[0195] 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 was no bitterness detected and 10 was similar to the maximum imaginable bitterness.

[0196] Result

[0197] The panelists rated the bitterness of Solution A as 4.33 ± 1.52, mean ± SD.

[0198] The panelists rated the bitterness of Solution B as 8.33 ± 1.52, mean ± SD.

[0199] These data indicate that the TAG composition containing butyrate moieties in infant formula has significantly less bitterness compared to tributyrin in infant formula.

[0200] Example 4 - Taste properties of 1,3 - dibutyryl - 2 - palmitoyl glycerol

[0201] 1,3 - Dibutyryl - 2 - palmitoyl glycerol (BPB) was synthesized as a single compound using the following synthesis:

[0202]

[0203] BPB was evaluated in a descriptive sensory panel assessment and was found to have a neutral taste and odor.

[0204] Example 5 - Digestion of triglycerides containing butyrate moieties

[0205] 5.1 Emulsion preparation

[0206] A 10 wt% oil - in - water emulsion stabilized by 0.3 wt% polyoxyethylene sorbitan monooleate ( Tween 80) was prepared by mixing Tween 80 into the oil phase at 40 °C and then mixing with the aqueous phase. An ultrasonic probe homogenizer was then used to generate the emulsion.

[0207] 5.2 Particle size determination

[0208] The droplet size of each lipid emulsion was measured by laser scattering using a Mastersizer 3000 equipped with Hydro SM from Malvern Instruments (Malvern, Worcestershire, United Kingdom). Emulsion particle size is cited as two values, the volume surface mean diameter D3,2 (D3,2 = 1 / 4∑nidi 3 / ∑nidi 2) or the volume length mean diameter D4,3 (D4,3 = 1 / 4∑nidi 4 / ∑nidi 3). Emulsion particle size results are the average of three measurements on two freshly prepared emulsions.

[0209] 5.3 In vitro digestion

[0210] A lipid emulsion (2 mL) containing 200 mg of fat was subjected to in vitro gastrointestinal lipolysis. Digestion was carried out in a thermostatic glass vessel (37 °C) in a pH-STAT setup controlled by a TIM 856 double burette pH-STAT (Radiometer Analytical, France). For gastric digestion, the sample was incubated with 8.5 mL of simulated gastric fluid (SGF) at 37 °C and a pH of 5.5 for 90 minutes. The simulated gastric fluid consisted of 150 mM NaCl, 450 U / mL pepsin, and 18 U / mL rabbit gastric lipase. Digestion was initiated by adding rabbit gastric lipase with an activity of 18 units of tributyrin per mL (TBU) (measured at pH 5.4).

[0211] The intestinal digestion step was carried out in a pH-stat where the pH was maintained constant at 6.8 by adding NaOH. A bile salt mixture (bile salts prepared in tris buffer) and a calcium solution (20 mM Ca, 176.5 mM tris, 150 mM NaCl) were added to the SGF sample mixture. The mixture was transferred to the pH-stat where the pH was adjusted to approximately 6.78. The intestinal digestion step started when the temperature reached 37 ± 0.5 °C. The pH was adjusted to pH 6.8, and after incubating for two minutes at this pH and temperature, a pancreatin solution was added. The final composition of the intestinal fluid was 10 mM CaCl2, 12 mM mixed bile salts, 0.75 mM phospholipids, 150 mM NaCl, and 4 mM tris(hydroxymethyl)aminomethane buffer. The intestinal digestion step was carried out for 3 hours in a titration manager from Radiometer. During the intestinal phase of digestion, the digestion kinetics were followed using pH-stat (TIM856, Radiometer) technology and expressed as titratable acid (instead of fatty acid), which can be calculated by the following formula:

[0212] TA = VNaOH ×0:05×1000

[0213] TA: Total titratable acid released, mmol; V NaOH : Volume of NaOH used to titrate the acid released within 3 h, mL.

[0214] 5.6 Result

[0215] Since the digestion of dietary lipids involves lipases from the stomach and intestine, the lipid digestibility was evaluated using the following 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 emulsified using polyoxyethylene sorbitan monooleate ( 80) and had similar particle size distributions and specific surface areas ( Figure 2 ), which means that the digestion differences mainly originated from the triglyceride molecular structure.

[0216] Figure 1 i) of A - C shows the digestion of tributyrin (C4), high - oleic sunflower oil (HOSFO, mainly C18:1), and triglycerides containing butyrate moieties according to the present invention generated by chemical transesterification between tributyrin and high - oleic sunflower oil (see Example 1) "C4 - C18:1" in the presence of mixed bile and calcium and by porcine pancreatic lipase (from pancreatin) (SIF model). Lipids generally exhibited the same lipolytic behavior, experiencing an initial rapid lipolysis period within the first 15 minutes and gradually slowing down during the last 2.5 hours of simulated intestinal digestion. The C4 triglyceride exhibited an initial maximum lipolysis rate of 223 ± 59 μmol.min -1 . The initial lipolysis rate of high - oleic sunflower oil was 34.5 ± 2.3 μmol.min -1 which was significantly lower (p < 0.0001) than that of short - chain triglycerides. C4 - C18:1 exhibited an initial hydrolysis rate of 153 ± 47 μmol.min -1 which was between the initial hydrolysis rates of C4 and C18:1. Overall, it can be seen that in the presence of porcine pancreatic lipase, all triglycerides were digested rapidly and extensively.

[0217] Next, triglycerides were digested using the sequential SGF(RGL)SIF(PPL) model, and the digestion in the SIF compartment is shown in Figure 1In ii) of A-C. Due to the limited ionization of the target fatty acids, measurements were not carried out in the gastric compartment. Compared with digestion with SIF alone, C4 and C18:1 triglycerides generally released a smaller amount of titratable acid during 3 hours of digestion. The effect was greatest with tributyrin, with an initial lipolysis rate of 223 ± 59 μmol.min -1 compared with an initial lipolysis rate of 44.1 ± 8.8 μmol.min during SGF-SIF digestion -1 which was significantly reduced (p < 0.0001). The total amount of acid released after SGF-SIF digestion of tributyrin, 381 ± 20 μmol, was almost one-third of the amount of acid released after SIF digestion alone, 958 ± 12.5 μmol. These results clearly show that there is extensive digestion of tributyrin in the gastric compartment of the model.

[0218] When exposed sequentially to SGF and SIF, the SIF lipolysis rate of C4-C18:1 triglycerides containing butyrate moieties was 124 ± 20 μmol.min -1 , showing a slight but non-significant decrease compared with SIF alone (124 ± 20 μmol.min -1 ). The most interesting observation was the effect of the secondary fatty acid chain length on the reduction in SIF lipolysis caused by pre-exposure to RGL. Initially, tributyrin showed a 60.2% reduction in total fatty acid release (147 ± 7.6 μmol) during SIF lipolysis after pre-exposure to RGL in SGF. In contrast, the C4-C18:1 transesterified triglyceride showed a 6.1% reduction (45 ± 7.6 μmol).

[0219] Figure 2 The overall extent of lipid digestion after SIF and SGF-SIF for three triglycerides using direct titration and back-titration is shown in. Since 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 back-titration to pH 11.5 or GC-FAME analysis is required to estimate the extent of complete digestion. The back-titration results for the three triglycerides showed that tributyrin and the C4-C18:1 triglyceride containing butyrate moieties underwent 101.5 ± 0.9% and 101 ± 1.6% digestion respectively, indicating that three fatty acids are released per molecule for complete digestion, while high oleic sunflower oil underwent 72.3 ± 2% digestion, indicating that two fatty acids are released per molecule for complete digestion.

[0220] Overall, 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 butyrate moieties generated by transesterification of C4 with long-chain fatty acids (C4-C18:1) reduce the degree of gastric lipolysis of C4 fatty acids. Tributyrin undergoes approximately 60% lipolysis by gastric lipase, as indicated by the decrease in total fatty acid release during SIF lipolysis after pre-exposure to RGL in SGF. In contrast, triglycerides with C4-C18:1 butyrate moieties showed only a 6.1% decrease in total fatty acid release in SGF-SIF. These results suggest that transesterification of C4 with long-chain fatty acids (C4-C18:1) regulates the release of butyric acid in the stomach and subsequently in the intestine after digestion, and that the design of structured lipids alters the timing (but not the extent) of short-chain fatty acid delivery in the gastrointestinal tract.

[0221] All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the methods, cells, compositions, and uses disclosed herein will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been disclosed in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications to the modes of practicing the invention disclosed herein that are apparent to those skilled in the art are intended to fall within the scope of the following claims.

Claims

1. A compound having the following formula, (1) or a combination thereof, which is used for preventing or treating cardiovascular disorders in an individual, wherein R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently long-chain fatty acids having 16 to 20 carbons.

2. A composition, the composition comprising a compound having the following formula (1) or a combination thereof, which is used for preventing or treating cardiovascular disorders in an individual, wherein R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are independently long-chain fatty acids having 16 to 20 carbons.

3. The composition for the use according to claim 2, wherein the composition comprises a compound having formula (1), a compound having formula (2), a compound having formula (3), and a compound having formula (4).

4. The composition for the use according to claim 2 or claim 3, wherein the compound having formula (1), the compound having formula (2), the compound having formula (3), and the compound having formula (4) account for at least 50% by weight, 60% by weight, 70% by weight, 80% by weight, 90% by weight, 95% by weight, or 99% by weight of the total triglycerides in the composition.

5. The composition for the use according to any one of claims 2 to 4, wherein the compound having formula (1), the compound having formula (2), the compound having formula (3), and the compound having formula (4) account for at least 50% by weight, 60% by weight, 70% by weight, 80% by weight, 90% by weight, 95% by weight, or 99% by weight of the triglycerides in the total butyrate-containing part of the composition.

6. The composition for the use according to any one of claims 2 to 5, wherein tributyrin accounts for less than 10% by weight, preferably less than 8% by weight, and more preferably less than 5% by weight of the total triglycerides in the composition.

7. The composition for the use according to any one of claims 2 to 6, wherein the composition further comprises dietary fiber.

8. A composition for the use according to any one of claims 2 to 7, wherein the composition is a nutritional composition, a dietary supplement or a pet care product.

9. A compound or combination thereof for the use according to claim 1, or a composition for the use according to any one of claims 2 to 8, wherein R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 is an unsaturated fatty acid, preferably monounsaturated.

10. A compound or combination thereof for the use according to claim 1 or claim 9, or a composition for the use according to any one of claims 2 to 9, wherein R 1 , R 2 , R 3 , R 4 , R 5 and / or R 6 is selected from oleic acid, palmitic acid or linoleic acid, preferably wherein R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are all oleic acid.

11. A compound or combination thereof for the use according to any one of claims 1, 9 or 10, or a composition for the use according to any one of claims 2 to 10, which is used for treating or preventing atherosclerosis, coronary heart disease, myocardial infarction, angina pectoris, stroke and / or heart failure.

12. A compound or combination or composition thereof for the use according to claim 11, which is used for preventing atherosclerosis.

13. A method for treating or preventing cardiovascular disorders, comprising administering to a human or animal individual an effective amount of a compound or combination thereof according to claim 1 or any one of claims 9 to 12, or a composition according to any one of claims 2 to 12.

14. The method for treating or preventing cardiovascular disorders according to claim 13, wherein the cardiovascular disorder is selected from atherosclerosis, coronary heart disease, myocardial infarction, angina pectoris, stroke and / or heart failure.

15. A compound or combination thereof for the use according to any one of claims 1 or 9 to 12, or a composition for the use according to any one of claims 2 to 12, or a method according to claim 13 or 14, wherein the compound or combination thereof has improved sensory properties relative to butyric acid, tributyrin and / or butyrate.

Images