Milk-derived lipids for reducing BCAA levels and related disorders

Abstract

Use of milk-derived lipids in reducing the risk of one or more of insulin resistance (IR), diabetes, cancer, in particular pancreatic cancer, obesity, heart failure, and cardiovascular disease (CVD), and / or reducing BCAA levels in a subject.

Description

Technical Field

[0001] The invention disclosed herein relates to the use of milk - derived lipids for reducing the branched - chain amino acid (BCAA) levels in a subject and optionally also for reducing one or more of the following risks in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD); to a composition comprising a lipid fraction for reducing the branched - chain amino acid (BCAA) levels in a subject and optionally also for reducing one or more of the following risks in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD). The invention further relates to a method for reducing the branched - chain amino acid (BCAA) levels in a subject and optionally also for reducing one or more of the following risks in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD), the method comprising the step of administering to the subject a composition comprising a lipid fraction. Background Art

[0002] Breastfeeding is the best way to ensure the healthy growth and development of infants in the first few months of life. The WHO recommends exclusive breastfeeding for the first six months of life, and thereafter the introduction of safe and appropriate complementary foods to supplement continued breastfeeding up to two years of age or longer. However, when a mother is unable or chooses not to breastfeed for some reason and a safe breastfeeding substitute is needed, breast milk substitutes produced according to strict international composition and safety standards have a legitimate role.

[0003] Branched - chain amino acids (BCAAs) play important nutritional and physiological functions in glucose and lipid metabolism, protein synthesis, and gut health and immunity. The catabolism and balance of BCAAs are closely related to health and disease, and some signaling pathways mediate BCAA metabolism and disease progression. In addition, BCAAs and their derivatives can be used as potential biomarkers for diseases such as insulin resistance (IR), type 2 diabetes (T2DM), cancer, and cardiovascular disease (CVD). A large number of publications have consistently demonstrated that the concentrations of branched - chain amino acids (BCAAs) in plasma and urine are associated with insulin resistance. (Nie et al., Int. J. Mol. Sci. 2018, 19, 954; doi: 10.3390 / ijms19040954).

[0004] Elevated serum levels of BCAA (especially Val and Leu) are considered as cardiometabolic risk markers, independent of body mass index (Sun, H. et al., Catabolic defect of branched-chain amino acids promotes heart failure. Circulation 2016, 133, 2038-2049).

[0005] Postprandial amino acid metabolism interacts with postprandial lipid metabolism and can synergistically promote metabolic disorders and even diseases. Studies have shown that abnormal lipid metabolism can lead to reduced catabolism of branched-chain amino acids (BCAAs), resulting in elevated levels of them in the blood, which is associated with metabolic disorders (such as insulin diseases) and can further promote metabolic diseases such as obesity and type 2 diabetes. Therefore, it is desirable to have low levels of BCAA and reduce the risk of related disorders such as metabolic diseases such as obesity and type 2 diabetes.

[0006] Elevated plasma BCAA levels are also associated with general diabetes, cancer (especially pancreatic cancer), and heart failure (Neinast M. et al., Annu Rev Physiol. February 10, 2019; 81: pp. 139-164. doi: 10.1146 / annurev-physiol-020518-114455).

[0007] The object of the present invention is to provide a composition that can better solve at least one of the above-mentioned desires and / or needs.

[0008] Studies have found that eating may cause an increase in BCAA levels. It is desirable to restore BCAA levels to their basal levels as quickly as possible to reduce one or more of the following risks: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD) or any other disorder associated with elevated BCAA levels. Surprisingly, it was found that this so-called postprandial response depends on the diet consumed. Even more surprisingly, it was found that BCAA returns to its basal level faster after consuming milk lipids (especially cream or MFGM) compared to consuming plant fats.

[0009] The object of the present invention is to provide an additional use of milk fat and / or milk lipids, provide an additional composition for use, and provide an additional method comprising the step of administering milk fat or milk lipids. The aim is to better solve at least one of the above-mentioned expectations and / or needs. Summary of the Invention

[0010] One or more objects of the present invention are achieved by the uses, compositions for use, and methods as defined in the claims. According to one aspect of the present invention, this is achieved by the features of claim 1. The present invention provides the use of milk-derived lipids for reducing the BCAA level in a subject; preferably, for reducing the postprandial BCAA level in a subject. Additionally and / or alternatively, according to another aspect of the present invention, there is provided a composition comprising a lipid fraction for reducing the BCAA level in a subject; wherein reducing the BCAA level in the subject is a reduction in the postprandial time required for the BCAA level to start decreasing after an initial increase after ingestion of the lipid fraction; wherein the amount of milk-derived lipids measured is at least 10 wt% relative to the total weight of the lipid fraction.

[0011] According to another aspect of the present invention, there is provided a method for reducing the BCAA level in a subject, the method comprising the step of administering to the subject a composition comprising a lipid fraction, wherein the amount of milk-derived lipids measured is at least 10 wt% relative to the total weight of the lipid fraction.

[0012] In one embodiment, the use, composition for use, and / or method according to the present invention are also used for reducing one or more of the following risks in a subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD). Detailed Description

[0013] In one aspect, the present invention relates to the use of a lipid fraction in reducing the BCAA levels in a subject, wherein reducing the BCAA levels in the subject is a reduction in the postprandial time required for the BCAA levels to start decreasing after an initial increase following ingestion of the lipid fraction, and wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and optionally, wherein the amount of milk-derived lipid as measured is at least 10% wt% relative to the total weight of the lipid fraction. In one embodiment, the present invention also relates to reducing one or more of the following risks in a subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD). In one embodiment, the present invention relates to the use of a lipid fraction in reducing one or more of the following risks and / or reducing the BCAA levels in a subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD), wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and wherein the amount of milk-derived lipid as measured is at least 10% wt% relative to the total weight of the lipid fraction.

[0014] As mentioned in different embodiments and aspects of the present invention, preferably, the amount of C15:0 in the lipid fraction is expressed as at least 0.20% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.12% by weight of the total fatty acids; more preferably, the amount of C15:0 in the lipid fraction is expressed as at least 0.30% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.18% by weight of the total fatty acids. As used herein, the amount of C15:0 in the lipid fraction refers to the amount of pentadecanoic acid esterified to the glycerol backbone, and likewise, the amount of C17:0 refers to the amount of heptadecanoic acid esterified to the glycerol backbone.

[0015] The term "treatment" with respect to a given disease or disorder includes, but is not limited to, inhibiting the disease or disorder, such as preventing the disease or disorder from progressing; alleviating the disease or disorder, such as causing the disease or disorder to regress; or alleviating the symptoms caused by or resulting from the disease or disorder, such as alleviating, preventing, or treating the symptoms of the disease or disorder.

[0016] The term "prevention" with respect to a given disease or disorder means preventing the development of the disease (if not yet affected), preventing the occurrence of the disease or disorder in a subject who may be predisposed to the disease or disorder but has not been diagnosed with the disease or disorder, and / or preventing the further development of the disease / disorder (if already affected).

[0017] Reduction of the risk of a disease or disorder means a decrease in the likelihood of development of a given disease or disorder (if not yet affected), and / or a decrease in the likelihood of further development of a given disease or disorder (if already affected), and / or a decrease in the likelihood of a given disease or disorder being diagnosed.

[0018] As used herein, "milk-derived lipid" consists of dairy fat (i.e., milkfat) and dairy phospholipid (i.e., milk phospholipid). Thus, the total lipid fraction consists of milk-derived lipid, vegetable fat, milk-derived phospholipid, and vegetable phospholipid. The "milk-derived lipid" used herein can be obtained from the milk of ruminants by methods known in the art, preferably from bovine milk fat, more preferably from cow milk fat. In principle, the milk fat source can be any available ruminant milk fat source, such as whole milk, cream, anhydrous milk fat (AMF), or milk fat fractions produced by dry fractionation, supercritical CO 2 extraction or other fractionation methods known in the art. The milk-derived lipid source can also be a source of milk fat globule membrane (MFGM). Similarly, the milk-derived lipid source can be a combination of different sources. However, it has been found particularly suitable to use whole milk, AMF, cream, and / or MFGM as the milk-derived lipid source. Preferably, the ruminant milk lipid is bovine milk lipid, and these lipids are selected from the group consisting of: anhydrous milk fat (AMF), cream, MFGM, and whole milk. More preferably, these lipids are selected from AMF, cream, or a mixture thereof. Even more preferably, these bovine milk lipids are cow milk lipids. In one embodiment, the milk-derived lipid is bovine whole milk or cream. In one embodiment, the milk-derived lipid is bovine whole milk; in another embodiment, the milk-derived lipid is milk oil; in still another embodiment, the milk-derived lipid is bovine AMF. Preferably, the milk-derived lipid source is bovine CREAM and / or bovine MFGM. As used herein, MFGM refers to a milk fraction rich in MFGM. Milk fractions rich in AMF and MFGM are readily commercially available from dairy processing companies such as FrieslandCampina, Arla, or Fonterra, e.g., VMF100 (FrieslandCampina), MFGM (FrieslandCampina), MFGM-10 (Arla), SureStart TMMFGM Lipids (NZMP). Conventional techniques can be applied to produce different milk lipid sources, such as whole milk, cream, anhydrous milk fat (AMF) or milk fat fractions produced by dry fractionation, critical CO 2 extraction or other fractionation methods. Such milk lipid sources are also readily commercially available. If a heat sterilization or pasteurization step is required (e.g., to meet legal requirements), methods known in the art can be used to achieve this, such as holding at a temperature of about 63 °C (145 °F) for 30 minutes, or heating to a higher temperature, such as 72 °C (162 °F) and holding for 15 seconds, or alternatively, for example, performing ultra-high temperature (UHT) treatment at a temperature of not less than 135 °C in combination with an appropriate holding time. Alternatively, ruminant milk lipids can be obtained from fresh milk.

[0019] As used herein, the term "cream" refers to the fat portion from whole milk (preferably bovine milk). This fat portion can be separated from whole milk using methods known in the art. Cream contains triglycerides and milk fat globule membrane (MFGM) fragments (such fragments may also be referred to as MFGM components). The MFGM fragments contain milk phospholipids and a variety of (glyco)proteins. Milk fat globules contain triglycerides surrounded by a membrane composed of MFGM fragments.

[0020] As used herein, "postprandial blood lipids" refers to the elevation of triglyceride-containing lipoproteins in the circulation after a meal, and postprandial blood lipids have been recognized as a risk factor for the development of cardiovascular diseases and other chronic diseases.

[0021] As used herein, the term "subject" preferably refers to a human. Unless the gender is explicitly specified, the term "subject" refers to both males and females. Human subjects can be infants (≤2 years old), juveniles, adolescents, adults or elderly subjects. In a preferred embodiment, the age of the human subject is from 0 to 36 months. In a particularly preferred embodiment of the present invention, the age of the human subject is 18 years old and above, such as at least 25 years old, at least 30 years old, at least 35 years old, at least 40 years old, or at least 45 years old. More preferably, the human subject is at least 50 years old, at least 55 years old, at least 60 years old, or at least 65 years old. Although there is no specific upper limit in practice, the human subjects treated according to the present invention are generally up to 110 years old at most, such as up to 100 years old or up to 90 years old at most.

[0022] As shown in the examples, the consumption of fat by humans results in an increase in BCAA levels. This increase is measured relative to the basal level. The basal level refers to the BCAA level of the subject in a fasting state for at least 10 hours. During the fasting state, the subject is not allowed to eat or drink, except for water. Thus, in different aspects and / or embodiments of the present invention, the reduction in BCAA levels refers to the reduction in the postprandial time required for the BCAA levels to start decreasing after an initial increase after the ingestion of the lipid fraction, preferably, after the subject consumes a composition containing the lipid fraction, the reduction in the postprandial time required for the BCAA levels to return to their basal level. As used herein, "postprandial time" is defined as the time after consuming a composition containing the lipid fraction. Preferably, the lipid fraction in the composition containing the lipid fraction mentioned in different aspects and / or embodiments of the present invention contains at least 10 wt% of milk-derived lipids (measured relative to the total weight of lipids in the lipid fraction).

[0023] Alternatively, in other embodiments of different aspects of the present invention, the reduction in BCAA levels is

[0024] i. compared to the postprandial time required for the BCAA levels of the subject to return to the basal level after consuming the same amount of vegetable oil, the postprandial time required for the BCAA levels of the subject to return to the basal level after consuming a composition containing the lipid fraction is reduced by at least 5%, preferably at least 10%, more preferably at least 15%, and most preferably at least 20%.

[0025] In other embodiments of different aspects of the present invention, the reduction in BCAA levels is ii. when the lipid fraction contains cream, between 5 and 6 hours postprandially, preferably between 4 and 6 hours postprandially, and most preferably between 3 and 6 hours postprandially, the BCAA levels of the subject are reduced.

[0026] In other embodiments of different aspects of the present invention, the reduction in BCAA levels is iii. the BCAA levels return to the basal level at 9 hours postprandially.

[0027] In other embodiments of different aspects of the present invention, the BCAA levels at 6 hours postprandially are 70% or lower, preferably 55% or lower, of the BCAA levels at 3 hours postprandially.

[0028] In other embodiments of different aspects of the present invention, the reduction in BCAA levels is v. the area under the curve (AUC) of the Δ postprandial response of BCAA is reduced by at least 10%, preferably at least 20%.

[0029] As used herein, the area under the curve (AUC) of the Δ postprandial response of BCAA refers to a similar Figure 2The curve in which the postprandial BCAA level minus the basal BCAA level is plotted as a function of time between t = 0 (basal level) and t = 6 hours.

[0030] In the compositions comprising a lipid fraction mentioned in different embodiments and / or aspects of the present invention, the amount of milk-derived lipid measured, relative to the total weight of the lipid fraction, can be at least 20 wt%, preferably at least 30 wt%, more preferably at least 50 wt%, particularly preferably at least 75 wt%, most preferably at least 90 wt%, for example at least 95 wt% or 99 wt%.

[0031] The reduced risk and / or reduced BCAA level of one or more of the following is preferably determined relative to a subject consuming a composition comprising a lipid fraction: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD), wherein the lipid fraction consists of vegetable oil rather than milk-derived lipid. For the avoidance of doubt, the amount of lipid in the consumed lipid fraction is the same. More preferably, the vegetable oil consists of a blend of palm olein, canola oil, palm kernel oil, and sunflower oil.

[0032] The compositions comprising a lipid fraction mentioned in different embodiments of the present invention may further comprise a carbohydrate fraction and a protein fraction; preferably, the composition comprising the lipid fraction is selected from the group consisting of infant nutritional products, adult nutritional products, sports nutritional products, beverages, confectionery, and food supplements. Such products are well known in the art and the amount of milk-derived lipid in such products can be increased by replacing other lipid sources using methods known in the art.

[0033] In another aspect, the present invention relates to a composition comprising a lipid fraction for reducing the BCAA level of a subject; wherein reducing the BCAA level of the subject is a reduction in the postprandial time required for the BCAA level to start decreasing after an initial increase upon ingestion of the lipid fraction; wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and optionally, wherein the amount of milk-derived lipid measured, relative to the total weight of the lipid fraction, is at least 10 wt%. In one embodiment, the composition for use in the present invention is also for reducing one or more of the following risks of a subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD).

[0034] In one embodiment, the present invention relates to a composition comprising a lipid fraction, the composition being for reducing one or more of the following risks in a subject and / or reducing the BCAA level in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD); wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and wherein the amount of milk-derived lipid measured is at least 10% wt% relative to the total weight of the lipid fraction. Preferably, reducing BCAA is such that the postprandial time required for the BCAA level to return to its basal level is reduced after the subject consumes the composition comprising the lipid fraction.

[0035] In yet another aspect, the present invention relates to a method for reducing the BCAA level in a subject, the method comprising the step of administering to the subject a composition comprising a lipid fraction, wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and optionally, wherein the amount of milk-derived lipid measured is at least 10% wt% relative to the total weight of the lipid fraction. In one embodiment, the method of the present invention is also for reducing one or more of the following risks in a subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD). In another embodiment, the present invention relates to a method for reducing one or more of the following risks in a subject and / or reducing the BCAA level in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD), the method comprising the step of administering to the subject a composition comprising a lipid fraction, wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and wherein the amount of milk-derived lipid measured is at least 10% wt% relative to the total weight of the lipid fraction. Preferably, reducing the BCAA level is such that the postprandial time required for the BCAA level to return to its basal level is reduced after the subject consumes the composition comprising the lipid fraction.

[0036] In yet another aspect, the present invention relates to the use of a lipid fraction, as defined in the different aspects and embodiments of the present invention, in the preparation of a medicament for reducing the BCAA level in a subject, the use comprising the step of administering to the subject a composition comprising the lipid fraction, wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; and optionally, wherein the amount of milk-derived lipid, as determined relative to the total weight of the lipid fraction, is at least 10% wt%.

[0037] In a preferred embodiment of the present invention, the reduction in the risk of diabetes is a reduction in the risk of type 2 diabetes (T2DM).

[0038] In one embodiment, the use, composition for use, and method of the present invention, as defined in the different aspects and embodiments of the present invention, relate to reducing one or more of the following risks in a subject and reducing the BCAA level in the subject: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD).

[0039] In another embodiment, the use, composition for use, and method of the present invention, as defined in the different aspects and embodiments of the present invention, relate to reducing the risk of insulin resistance (IR) in a subject and / or reducing the BCAA level in the subject. In yet another embodiment, the present invention relates to reducing the risk of diabetes in a subject and / or reducing the BCAA level in the subject. In still another embodiment, the present invention relates to reducing the risk of cancer (especially pancreatic cancer) in a subject and / or reducing the BCAA level in the subject. In another embodiment, the present invention relates to reducing the risk of obesity in a subject and / or reducing the BCAA level in the subject. In another embodiment, the present invention relates to reducing the risk of heart failure in a subject and / or reducing the BCAA level in the subject. In still another embodiment, the present invention relates to reducing the risk of cardiovascular disease in a subject and / or reducing the BCAA level in the subject. Alternatively, the present invention relates to reducing the BCAA level in a subject.

[0040] In a preferred embodiment, the BCAA level, as mentioned in the various aspects and embodiments of the present invention, is the leucine level.

[0041] In a particularly preferred embodiment, the BCAA level, as mentioned in the various aspects and embodiments of the present invention, is the leucine level, and the lipid fraction comprises CREAM or MFGM.

[0042] It should also be understood that the present invention is not limited to the specific embodiments and methods described herein, as the specific components and / or conditions can of course vary. In addition, the terms used herein are for the purpose of describing specific embodiments of the present invention only and are not intended to be limiting in any way.

[0043] It must also be noted that, as used in the specification and the appended claims, the singular forms "a / an" and "the" include plural referents unless the context clearly dictates otherwise. For example, a component recited in the singular is intended to include a plurality of components.

[0044] It will be understood that in this disclosure, any reference to weight, weight ratio, etc. relates to dry matter, particularly to the dry matter of the composition.

[0045] Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0046] As used herein, the term "comprising" is synonymous with "including" or "containing", is open-ended, and does not exclude other unrecited elements, ingredients or method steps; while the term "consisting of" is a closed term and does not include any other elements, steps or ingredients not expressly recited.

[0047] As used herein, the term "essentially consisting of" is a partially open-ended term and does not exclude additional unrecited one or more elements, one or more steps, or one or more ingredients, provided that these additional one or more elements, one or more steps or one or more ingredients do not materially affect the basic and novel characteristics of the present invention.

[0048] As used herein, the term "comprising" includes the terms "consisting of" and "essentially consisting of". Thus, in this application, the term "comprising" means more specifically encompassing the terms "consisting of" and "essentially consisting of".

[0049] Throughout this application, where publications are cited, the disclosures of these publications are hereby incorporated by reference in their entirety to more fully describe the state of the art to which this invention pertains.

[0050] Except as otherwise indicated in the examples or otherwise specifically noted, all numerical values indicating amounts of materials or reaction and / or use conditions in the specification should be understood to be modified by the word "about" when describing the broadest scope of the present invention. It is generally preferred to practice within the stated numerical limits. Additionally, unless specifically stated to the contrary: percentages, "parts", and ratio values are by weight; the description of a group or class of materials suitable for or preferred for a given purpose related to the present invention means that a mixture of any two or more components of that group or class may equally be suitable or preferred; the description of a component in chemical terms refers to the component as added to any combination specified in the specification and does not necessarily exclude chemical interactions between the components of the mixture once mixed; the first definition of an acronym or other abbreviation applies to all subsequent uses of the same abbreviation in the present text, and the necessary modifications are applied to the normal grammatical variations of the initially defined abbreviation; and, unless specifically stated to the contrary, the measurement of a property is determined by the same technique as the same property cited previously or subsequently.

[0051] In the following, the present invention will be illustrated with reference to the following non-limiting examples. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 Different effects of interventions on postprandial metabolites. A) Study design and B) Flow chart of metabolites with significant changes during the intervention. Repeated measurements were performed using a linear mixed model to test the milkshake type * time effect. The significant results of multiple tests were corrected using FDR correction (FDR p-value < 0.05). Post hoc LSD tests were performed on metabolites with significant FDR p-values for milkshake type * time to determine the differences between milkshake types and milkshake type * time effects at specific time points.

[0053] Figure 2 Figure 2 a, Δ postprandial response of total branched-chain amino acids (BCAAs) (i.e., leucine, isoleucine, and valine ( Figure 2 a), in mmol / 1). All values are expressed as mean + SD. Milkshake type * time effect in the linear mixed model analysis (FDR p-value = 0.001). Significant difference between VEGE and CREAM (p = 0.018).

[0054] Figure 2 b, Δ postprandial response of leucine (in mmol / l) ( Figure 2 b). Milkshake type * time effect in the linear mixed model analysis (FDR p-value = 0.001). Significant difference between VEGE and AMF (p = 0.026), and significant difference between VEGE and CREAM (p < 0.001).

[0055] a, b, and c represent significant differences in milkshake type and milkshake type * time effect at specific time points. All values are expressed as mean + SD.

[0056] Example Example 1

[0057] Fatty acid determination

[0058] The content of different fatty acids in the lipids of the compositions of the present invention can be determined by standard methods ISO 15884 / IDF 182:2002 (Milk fat - Preparation of fatty acid methyl esters) and ISO 15885 / IDF 184 (Milk fat - Determination of fatty acid composition by gas - liquid chromatography). These ISO methods allow the determination of the molar concentration of fatty acids relative to the total molar number of that fatty acid in TAG ([FA - TAG]). The fatty acid distribution on the glycerol backbone can be determined according to the method disclosed in the official JOCS / AOCS method Ch 3a - 19: Enzymatic transesterification method using Candida antarctica lipase for the determination of the fatty acid composition at the 2 - position of oils and fats (approved in 2019). Essentially, this method involves hydrolyzing triacylglycerol (TAG) using sn - 1,3 - specific pancreatic lipase (porcine). The required 2 - monoacylglycerol formed is separated by thin - layer chromatography and then methylated for gas chromatography analysis and quantified in molar concentration relative to the total molar number of fatty acids at the sn - 2 position ([FA(sn - 2)]).

[0059] Determine the branched - chain amino acid (BCAA) level of the subject

[0060] The amino acid levels, including total BCAA and leucine, were measured using the Nightingale Health platform (https: / / research.nightingalehealth.com / blood-biomarker-analysis section), as described by Würtz et al. (P. Würtz et al. Quantitative Serum Nuclear magnetic Resonance metabonomics in Large-Scale Epidemiology: A Primer on - Omics Technologies [Quantitative Serum Nuclear Magnetic Resonance Metabonomics in Large-Scale Epidemiology: An Introduction to Omics Technologies], Am. J. Epidemiol [American Journal of Epidemiology] 2017, Vol. 186(9), pp. 1084 - 1096). Briefly, a high-throughput proton (1H) nuclear magnetic resonance (NMR) metabonomics method was used, as described by Soininen et al. (Soininens et al. High-throughput serum NMR metabonomics for cost-effective holistic studies on systemic metabolism [High-Throughput Serum NMR Metabonomics for Cost-Effective Holistic Studies of Systemic Metabolism]; Analyst [Analyst], 2009, Vol. 134, pp. 1781 - 1785).

[0061] Clinical research

[0062] A study was conducted to investigate the postprandial responses of plasma metabolites to the consumption of high-fat milkshakes, including plant fats and bovine milk fats with or without milk fat globule membrane.

[0063] Study population

[0064] The study population consisted of 40 individuals aged 40 - 70 years with a BMI of 22 - 27 kg / m 2Consisted of healthy men and women. Participants were excluded if they had any chronic metabolic diseases, gastrointestinal diseases, inflammatory diseases, or other chronic diseases, if they had gastrointestinal discomfort or surgery (or a history of gastrointestinal disease or surgery), renal or liver dysfunction (measured by ALAT, ASAT, and creatinine), or if they were using medications that could affect the study results by influencing intestinal motility. Additionally, participants were excluded if they were pregnant (intending to become pregnant), using soft / hard drugs, drinking alcohol >14 times per week, smoking, or had unstable body weight. Finally, participants were excluded from the study if they followed a vegan diet or had a food allergy to any of the products used in the study. All participants provided written consent before participating in the study.

[0065] Study design

[0066] This study was a double-blind, randomized acute intervention study (Figure 1a). Each participant visited the research institution three times, with a washout period of at least one week. During each study visit, participants underwent a dietary lipid load challenge test in the form of a high-fat milkshake. Participants were randomly assigned to a series of milkshakes.

[0067] The night before each study visit, participants consumed a standardized meal (ad libitum) and were not allowed to eat or drink anything except water until the next day. The next day, participants appeared at the research institution in a fasting state of at least 10 hours.

[0068] This study was conducted at Wageningen University in the Netherlands from July 1, 2020, to October 3, 2020. The experimental protocol and procedures were approved by the Medical Ethics Committee of Wageningen University and complied with the Declaration of Helsinki as adopted in 1975 (revised in 1983). This study has been registered on clinicaltrials.gov as NCT04178681 "Postprandial Effects of Milk Fat (POEMI)".

[0069] Dietary lipid load challenge test

[0070] During each study visit, participants underwent a dietary lipid load challenge test. In this test, participants consumed a liquid milkshake containing skim milk (0.5 L) and 95 grams of fat from different sources (Table 1).

[0071] Table 1. Nutritional information of the high-fat milkshake

[0072] Amount per 618-gram milkshake (consisting of water, skim milk, and fat milkshake powder).

[0073]

[0074] *0.5 g of fat from skim milk

[0075] The milkshakes were prepared by dissolving the fat blend in 500 ml of skim milk.

[0076] During this study, three types of fat were consumed, namely: 100% vegetable fat blend (VEGE), 100% anhydrous milk fat (AMF, bovine milk fat), and 100% cream (CREAM) (AMF + milk fat globule membrane). These milkshakes were isocaloric, with the only difference being the fat source, and the remaining ingredients were the same. The fats were provided by FrieslandCampina in powder form (Table 2). The milkshakes were prepared by dissolving the fat powder in skim milk.

[0077] Table 2. Composition table of three different fat powders

[0078]

[0079] Fatty acid composition of the lipid blend

[0080] The fatty acid compositions of the different fat blends given in Table 3 were determined using the method described elsewhere in this example. The amounts listed are expressed as weight percentages (w / w%) of the total fatty acids in the lipid fraction.

[0081] Table 3 Fatty acid composition of different fat milkshakes

[0082] Fatty acid (w / w%) * VEGE AMF Cream C15：0 ＜0.1 1.1 1.1 C17：0 ＜0.1 0.5 0.5

[0083] *The amounts listed are expressed as weight percentages (w / w%) of the total fatty acids in the lipid fraction.

[0084] The milkshakes were served to the participants in colored cups equipped with opaque straws to conceal the contents. The participants had to consume the milkshakes within ten minutes, unless the final blood sample was drawn, and were not allowed to eat or drink anything except water (ad libitum). During this period, the participants were also not allowed to engage in physical exercise.

[0085] Blood collection

[0086] During each study visit, a catheter cannula was inserted into the antecubital vein of the participant. Thirty minutes after the insertion of the cannula, blood was drawn from the catheter cannula for baseline measurement (t = 0). After this baseline measurement, the participants consumed the high-fat milkshakes. Subsequently, blood was drawn at t = 1, 2, 3, 4, 5, 6, 7, and 8 hours after consumption. All blood samples were stored at -80 °C until further analysis.

[0087] Metabolomics

[0088] Blood samples collected at 0 hours (baseline), 3 hours, and 6 hours were processed by Nightingale Health. The platform performs high-throughput proton nuclear magnetic resonance (NMR). Details of this analysis have been described previously (Soininen, P., Kangas, A.J., Würtz, P., Suna, T., and Ala-Korpela, M. (2015). Quantitative serum nuclear magnetic resonance metabolomics in cardiovascular epidemiology and genetics. [Quantitative serum nuclear magnetic resonance metabolomics in cardiovascular epidemiology and genetics] Circ. Cardiovasc. Genet. [Cardiovascular Circulation Genetics] 8, 192-206; Würtz, P., Kangas, A.J., Soininen, P., Lawlor, D.A., Davey Smith, G., and Ala-Korpela, M. (2017). Quantitative Serum Nuclear Magnetic Resonance Metabolomics in Large-Scale Epidemiology: A Primer on - Omic Technologies. [Quantitative Serum Nuclear Magnetic Resonance Metabolomics in Large-Scale Epidemiology: A Primer on - Omic Technologies] Am. J. Epidemiol. [American Journal of Epidemiology] 186, 1084-1096); Soininen et al. High-throughput serum NMR metabonomics for cost-effective holistic studies on systemic metabolism [High-throughput serum NMR metabonomics for cost-effective holistic studies on systemic metabolism]; Analyst [Analyst], 2009, Vol. 134, pp. 1781-1785). This analysis allows the simultaneous detection and quantification of 249 metabolites and their corresponding ratios in a single experimental setup. These include lipoprotein subclasses and their components, their relative ratios, lipids, fatty acids, amino acids, ketone bodies, glycolysis-related metabolites, and various other low-molecular-weight metabolites.

[0089] Statistical analysis

[0090] Metabolites were statistically analyzed using log2-transformed data. A linear mixed model with repeated measures was used to analyze the postprandial response differences among the three different milkshakes in the dietary lipid load test. Participants were set as the subjects. Milkshake type and time were set as repeated measures factors. The Δ values (t = 3 h - t = 0 h and t = 6 h - t = 0 h) of 249 metabolites were selected as the dependent variables in the model. Milkshake type, time, and the interaction of milkshake type * time were all included as fixed effects. This model selected a first-order autoregressive covariance structure. An additional LSD post hoc test was performed on metabolites with a significant milkshake type * time effect to determine the differences between milkshake types at specific time points and specific milkshake type * time effects. The Benjamini-Hochberg FDR correction method was used to correct for multiple testing of the results of the linear mixed model (FDR p-value < 0.05) ([Benjamini, Y. and Hochberg, Y. (1995) Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing. J. R. Stat. Soc. Ser. B 57, 289 - 300). All statistical analyses were performed using Statistics (version 28.0.1.0). Figures were created in GraphPad Prism (version 9.3.1) and Cytoscape (version 3.9.1).

[0091] Results

[0092] Participant characteristics

[0093] Among the 40 participants involved in this study, 37 completed the study. One participant withdrew after the first study visit, and two other participants withdrew after the second study visit ( Figure 1A ). The data of the withdrawers were still included in the analysis. The characteristics of the 40 participants are shown in Table 4.

[0094] Table 4. Characteristics of the 40 participants included in the study

[0095] Data are presented as mean ± standard deviation.

[0096]

[0097]

[0098] Abbreviations: alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), body mass index (BMI), hemoglobin (Hb), waist-to-hip ratio (WHR)

[0099] AMF, CREAM, and VEGE had the same effect on postprandial metabolites

[0100] The effects of three fat sources on postprandial plasma metabolite levels were investigated (comparison between shakes). Among the 249 metabolites measured in total, 102 metabolites had different changes in response to the three shakes postprandially. Post hoc tests were performed to determine differences between the shakes at specific time points. Three hours postprandially, 37 metabolites showed significant differences between VEGE and AMF, 39 metabolites showed significant differences between VEGE and CREAM, and no metabolites showed significant differences between AMF and CREAM. Six hours postprandially, 41, 29, and 8 metabolites were determined to have significant differences in the above comparisons, respectively (Figure 1b).

[0101] Significant changes in metabolites in the comparison between shakes and within shakes

[0102] Repeated measures of the differences between the three shakes were performed using a linear mixed model. Among 239 metabolites, 102 metabolites had a significant shake type * time effect. The significant results of multiple tests were corrected using FDR correction (FDR p-value < 0.05). Additional LSD post hoc tests were performed to determine differences between shake type and shake type * time effect at specific time points (Figure 1b).

[0103] Consumption of AMF and CREAM enabled a more rapid decline in the postprandial elevation of the marker BCAA

[0104] Branched-chain amino acids (BCAAs) play important nutritional and physiological functions in glucose and lipid metabolism, protein synthesis, and gut health and immunity. BCAA levels are closely associated with health and diseases such as insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD). Surprisingly, there were significant differences in the postprandial responses of BCAA among the three shakes. Specifically, compared with VEGE, CREAM consumption led to lower levels of BCAA and leucine 6 hours postprandially. CREAM consumption caused the postprandial elevation of BCAA to rapidly return to its basal level, while this did not occur with VEGE consumption ( Figure 2 a and Figure 2b). As used herein, "BCAA level" refers to the Δ value of the BCAA level measured at a specific post - meal time point compared to the BCAA levels at baseline (t = 3h - t = 0h and t = 6h - t = 0h). Three hours after the meal, CREAM has the highest BCAA level, while the BCAA level of VEGE reaches its peak increase only at 6 hours after the meal. It is further noted that at t = 6, the BCAA level of CREAM has already returned to 50% of its level at t = 3.

[0105] It was observed that 6 hours after the meal following CREAM consumption, the post - meal increase in branched - chain amino acids and leucine was smaller and had almost returned to baseline, while this indicator for VEGE consumption was still increasing at 6 hours after the meal.

[0106] When comparing AMF with VEGE, a similar but weaker effect was observed. At 3 hours and 6 hours after the meal, the BCAA and leucine levels after AMF consumption were lower compared to VEGE. The maximum BCAA and leucine levels after AMF consumption were still lower compared to VEGE consumption. It was observed that from post - meal t = 3 to t = 6, the increase in BCAA and leucine for AMF was much lower compared to VEGE.

[0107] Based on these results, the lipid fraction in different aspects and embodiments of the present invention preferably comprises CREAM or MFGM, more preferably comprises CREAM, and particularly preferably comprises MFGM. Most preferably, the lipid fraction consists of CREAM or MFGM.

[0108] Example 2

[0109] Table 5 below gives an example of a nutritional composition (such as an infant formula) suitable for different embodiments or aspects of the present invention.

[0110] Table 5:

[0111]

[0112] * Whey protein concentrate (SPC) is a protein present in cheese whey and can be directly extracted from milk. Since SPC does not go through the cheese - making process, there are fewer enzymatic or chemical reactions that can cause off - flavors.

[0113] Example 3

[0114] An example of a nutritional composition (such as an adult nutritional product) suitable for different embodiments or aspects of the present invention is given below:

[0115] Ingredients: water, milk protein (SPC), glucose, vegetable oils (sunflower oil, rapeseed oil), milk fat, sucrose, fibers (fructooligosaccharides, inulin, galactooligosaccharides), minerals, fish oil, emulsifiers, vitamins, carnitine, stabilizers, flavoring agents, taurine.

[0116] The edible portion of the composition per 100.0 ml contains:

[0117]

[0118]

Claims

1. Use of a lipid fraction in reducing the branched-chain amino acid (BCAA) level in a subject, wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids; wherein reducing the BCAA level in the subject is a reduction in the postprandial time required for the BCAA level to start decreasing after an initial increase after ingestion of the lipid fraction; preferably, wherein reducing the BCAA level is determined in the subject consuming the composition comprising the lipid fraction, wherein the lipid fraction consists of vegetable oil rather than milk-derived lipid.

2. Use according to claim 1, wherein reducing the BCAA is a reduction in the postprandial time required for the BCAA level to return to its basal level after the subject consumes the composition comprising the lipid fraction.

3. Use according to any one of the preceding claims, wherein the lipid fraction comprises at least 10 wt% of milk-derived lipid, determined relative to the total weight of the lipids in the lipid fraction.

4. A composition comprising a lipid fraction for reducing the BCAA level in a subject; wherein reducing the BCAA level in the subject is a reduction in the postprandial time required for the BCAA level to start decreasing after an initial increase after ingestion of the lipid fraction; and wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids.

5. The composition for use according to claim 4, wherein reducing the BCAA is a reduction in the postprandial time required for the BCAA level to return to its basal level after the subject consumes the composition comprising the lipid fraction.

6. The composition for use according to claim 4 or 5, wherein the lipid fraction comprises at least 10 wt% of milk-derived lipid, determined relative to the total weight of the lipids in the lipid fraction.

7. A method for reducing the BCAA level in a subject, the method comprising the step of administering to the subject a composition comprising a lipid fraction, wherein the amount of C15:0 in the lipid fraction is expressed as at least 0.10% by weight of the total fatty acids, and the amount of C17:0 in the lipid fraction is expressed as at least 0.06% by weight of the total fatty acids.

8. The method according to claim 7, wherein reducing the BCAA level is a reduction in the postprandial time required for the BCAA level to return to its basal level after the subject consumes the composition comprising the lipid fraction.

9. The method according to claim 7 or 8, wherein the amount of milk-derived lipid determined relative to the total weight of the lipid fraction is at least 10 wt%.

10. Use, composition for use, or method according to any one of the preceding claims, wherein the composition comprises a lipid fraction, and the amount of milk-derived lipid, as measured relative to the total weight of the lipid fraction, is at least 20 wt%, preferably at least 30 wt%, more preferably at least 50 wt%, particularly preferably at least 75 wt%, most preferably at least 90 wt%, such as at least 95 wt% or 99 wt%.

11. Use, composition for use, or method according to any one of the preceding claims, wherein the milk-derived lipid is selected from one or more of anhydrous milk fat (AMF), cream, MFGM, and whole milk; preferably, the milk-derived lipid is selected from AMF, cream, or a mixture thereof.

12. Use, composition for use, or method according to any one of the preceding claims, wherein the subject is a human subject, preferably a human subject aged 0 to 36 months, particularly preferably a human subject aged 18 years or older, more preferably a human subject aged 50 years or older.

13. Use, composition for use, or method according to any one of the preceding claims, wherein the composition comprising the lipid fraction further comprises a carbohydrate fraction and a protein fraction; preferably, the composition comprising the lipid fraction is selected from the group consisting of infant nutrition products, adult nutrition products, sports nutrition products, beverages, confectionery, and food supplements.

14. Use, composition for use, or method according to any one of the preceding claims, wherein i. the postprandial time required for the BCAA level of the subject to return to the basal level after consuming the composition comprising the lipid fraction is reduced by at least 5%, preferably at least 10%, compared to the postprandial time required for the BCAA level of the subject to return to the basal level after consuming the same amount of vegetable oil; and / or ii. when the lipid fraction comprises cream, the BCAA level of the subject decreases between 5 and 6 hours postprandially.

15. Use, composition for use, or method according to any one of the preceding claims, wherein the BCAA level is reduced as defined therein, and wherein one or more of the following risks are reduced for the subject consuming the composition comprising the lipid fraction: insulin resistance (IR), diabetes, cancer (especially pancreatic cancer), obesity, heart failure, and cardiovascular disease (CVD), wherein the lipid fraction consists of vegetable oil rather than milk-derived lipid.

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