WO2018102914A1

WO2018102914A1 - Use of medium-chain triglycerides for the management of metabolic conditions

Info

Publication number: WO2018102914A1
Application number: PCT/CA2017/051463
Authority: WO
Inventors: Catherine MOUNIER; Sabri RIAL
Original assignee: Transfert Plus, Société En Commandite
Priority date: 2016-12-06
Filing date: 2017-12-05
Publication date: 2018-06-14

Abstract

The use of a composition comprising an effective amount of medium-chain triglycerides (MCT) or medium-chain fatty acids (MCFA), such as caprylic acids and/or capric acids, for preventing or reducing a metabolic condition selected from insulin resistance, glucose intolerance, non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH) in a subject, is disclosed. The composition may be in the form of a food composition or food supplement, or a pharmaceutical composition.

Description

USE OF MEDIUM-CHAIN TRIGLYCERIDES FOR THE MANAGEMENT OF

METABOLIC CONDITIONS

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefits of U.S. Provisional serial No. 62/430,593, filed December 6, 2016, which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

The present invention generally relates to metabolic conditions, and more particularly pre-diabetic conditions such as fatty liver diseases as well as insulin and glucose resistance.

BACKGROUND ART

Non-alcoholic fatty liver disease (NAFLD), is the most common chronic liver disease in

North America. It is estimated that about 10 to 30 percent of Americans suffer from NAFLD. Several of these patients show no inflammation or liver damage, but others develop a condition called non-alcoholic steatohepatitis (NASH) which is a clinical syndrome of steatosis accompanied by hepatic inflammation and affects about 2 to 5 percent of Americans. NAFLD starts out as ectopic lipid deposition in extra-adipose tissues, such as liver. The lipid deposition raises oxidative stress in the affected cells and induces an inflammatory response. Fatty liver disease begins with ectopic lipid deposition in hepatocytes, steatosis, progressing to inflammation (NASH), then to fibrosis cirrhosis and ultimately to hepatocellular carcinoma or liver failure. No specific cause of NAFLD or NASH is known, but it is commonly associated with conditions such as obesity, insulin resistance, diabetes, dyslipidemia (e.g., excess of blood triglycerides and/or cholesterol), thyroid disorders or hypertension.

Insulin resistance is characterized by a poor response of skeletal muscles, adipose tissues, and liver to insulin, and in turn to poor absorption of glucose from the bloodstream. This typically leads to increased glycemia, then to increased secretion of insulin by β-cells in the pancreas to compensate for the higher levels of insulin needed to help glucose enter cells. Over time, insulin resistance can lead to pre-diabetes (notably impaired glucose tolerance (IGT) or glucose intolerance) and type 2 diabetes due to the failure of the β-cells to keep up with the body's increased need for insulin.

Although some drugs such as metformin can delay or prevent the onset of diabetes, lifestyle modifications play a significant role in the prevention of type 2 diabetes. Patients identified as having a pre-diabetic state such as insulin resistance or IGT may be able to prevent or delay diabetes through a combination of increased exercise and reduction of body weight. There is a need for the development of novel approach for preventing, reducing and/or treating metabolic conditions, and more particularly pre-diabetic conditions such as insulin resistance, IGT, as well as other metabolic conditions such as NAFLD or NASH.

The present description refers to a number of documents, the content of which is herein incorporated by reference in their entirety.

SUMMARY OF THE INVENTION

The present invention provides the following items

1. A method for preventing or reducing a metabolic condition in a subject, said method comprising administering a composition comprising an effective amount of medium-chain triglycerides (MCT) or medium-chain fatty acids (MCFA) to said subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, non-alcoholic fatty liver disease (NAFLD) or non-alcoholic hepatic steatosis (NASH).

2. The method of item 1 , wherein said metabolic condition is insulin resistance.

3. The method of item 1 or 2, wherein said metabolic condition is glucose intolerance. 4. The method of any one of items 1 to 3, wherein said metabolic condition is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

5. The method of any one of items 1 to 4, wherein said composition comprises at least about 5% of MCT or MCFA (wt/wt of the composition).

6. The method of any one of items 1 to 4, wherein said composition comprises at least about 10% of MCT or MCFA (wt/wt of the composition).

7. The method of any one of items 1 to 4, wherein said composition comprises about 5% to about 50% of MCT or MCFA (wt/wt of the composition).

8. The method of any one of items 1 to 4, wherein said composition comprises about 10% to about 30% of MCT or MCFA (wt/wt of the composition).

9. The method of any one of items 1 to 8, wherein said composition is administered prior to the development of said metabolic condition.

10. The method of any one of items 1 to 9, wherein said composition is administered at least once daily.

1 1. The method of any one of items 1 to 10, wherein said MCT or MCFA comprise C8 fatty acids (caprylic acids).

12. The method of any one of items 1 to 1 1 , wherein said MCT or MCFA comprise C10 fatty acids (capric acids).

13. The method of any one of items 1 to 12, wherein said MCT or MCFA comprise (i) caprylic acids and (ii) capric acids.

14. The method of item 13, wherein said MCT or MCFA comprise at least 40% of caprylic acids and at least 40% of capric acids. 15. The method of any one of items 1 to 14, wherein said composition is a food composition or food supplement.

16. The method of any one of items 1 to 15, wherein said composition is a concentrated MCT or MCFA oil composition.

17. The method of any one of items 1 to 14, wherein said composition is a pharmaceutical composition.

18. Use of a composition comprising an effective amount of medium-chain triglycerides (MCT) or medium-chain fatty acids (MCFA) for preventing or reducing a metabolic condition in a subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, non-alcoholic fatty liver disease (NAFLD) or non-alcoholic hepatic steatosis (NASH).

19. Use of a composition comprising an effective amount of medium-chain triglycerides (MCT) or medium-chain fatty acids (MCFA) for the manufacture of a medicament for preventing or reducing a metabolic condition in a subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, non-alcoholic fatty liver disease (NAFLD) or nonalcoholic hepatic steatosis (NASH).

20. The use of item 18 or 19, wherein said metabolic condition is insulin resistance.

21. The use of item 18 or 19, wherein said metabolic condition is glucose intolerance.

22. The use of any one of items 18 to 21 , wherein said metabolic condition is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic hepatic steatosis (NASH).

23. The use of any one of items 18 to 22, wherein said composition comprises at least about 5% of MCT or MCFA (wt/wt of the composition).

24. The use of any one of items 18 to 22, wherein said composition comprises at least about 10% of MCT or MCFA (wt/wt of the composition).

25. The use of any one of items 18 to 22, wherein said composition comprises about 5% to about 50% of MCT or MCFA (wt/wt of the composition).

26. The use of any one of items 18 to 22, wherein said composition comprises about 10% to about 30% of MCT or MCFA (wt/wt of the composition).

27. The use of any one of items 18 to 26, wherein said composition is administered prior to or after the development of said metabolic condition.

28. The use of any one of items 18 to 27, wherein said composition is administered at least once daily.

29. The use of any one of items 18 to 28, wherein said MCT or MCFA comprise C8 fatty acids (caprylic acids).

30. The use of any one of items 18 to 29, wherein said MCT or MCFA comprise C10 fatty acids (capric acids). 31. The use of any one of items 18 to 30, wherein said MCT or MCFA comprise (i) caprylic acids and (ii) capric acids.

32. The use of item 31 , wherein said MCT or MCFA comprise at least 40% of caprylic acids and at least 40% of capric acids.

33. The use of any one of items 18 to 32, wherein said composition is a food composition or food supplement.

34. The use of any one of items 18 to 33, wherein said composition is a concentrated MCT or MCFA oil composition.

35. The use of any one of items 18 to 32, wherein said composition is a pharmaceutical composition.

Other objects, advantages and features of the present invention will become more apparent upon reading of the following non-restrictive description of specific embodiments thereof, given by way of example only with reference to the accompanying drawings.

BRIEF DESCRIPTION OF DRAWINGS

In the appended drawings:

FIG. 1A shows the detailed composition of the diets used in the experiments described herein.

FIG. 1 B is a schematic of the caloric properties of the different diets used in the experiments described herein.

FIG. 1C is a schematic of the protocol used to assess the curative effect of MCT in obese mice. 32 C57BL6/J mice (5-week-old) were first fed with a HFD for 10 weeks to induce obesity. The obese mice were then split into 4 groups of 8 mice, which were fed for an additional 10 weeks with the four diets described in Example 1 (LFD, HFD, MCT-20 and MCT- 40). The glucose tolerance test (GTT) and the insulin tolerance test (ITT) were performed at weeks 23 and 24, respectively. Mice were sacrificed at week 25 to assess liver parameters.

FIGs. 2A and 2B are graphs showing the effects of the different diets on energy intake and body weight, respectively. Four groups of 6-week-old C57BL6/J mice (n=8 per group) were fed during 10 weeks with the four diets described in Example 1. FIG. 2A: daily food intake was measured and converted to energy consumed. FIG. 2A shows the area under curve (AUC) of this daily energy intake, hence the cumulative energy consumed during the 10-week diet period for each group (n=8). FIG. 2B: AUC of the weekly weight gain, hence the cumulative weight gained during the 10-week diet period for each group (n=8). The initial weight of mice was comparable in each group. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post-hoc Tukey tests. Differences were considered significant when p-value < 0.05.

FIG. 2C shows the weekly weight gain in 5-week-old mice (32 males, C57BL/6J background) submitted to HFD during 10 weeks. FIGs. 2D and 2E are graphs showing the area under curve (AUC) of the daily food (FIG. 2D) and energy (FIG. 2E) intake during the 10-week diet period for each group (LFD, HFD, MCT-20 and MCT-40, n=8) of mice previously fed with a HFD for 10 weeks to induce obesity.

FIG. 2F is a graph showing the body weight changes in obese mice over the 10-week treatment with the LFD, HFD, MCT-20 and MCT-40 diets, relative to the baseline. Body weight was measured weekly for each mouse (n=8 per group) and converted into percentage of weight measured at week 15 (100% = weight after the 10-week HFD diet, week 15).

FIGs. 2G and 2H are graphs showing the AUC of the weekly weight gain data depicted in FIG. 2F. FIG. 2H shows the AUC of the weekly weight gain compared with the AUC of the theoretical baseline. Data is presented as average ± standard error. Results from the 4 groups were compared using an unpaired Student t-test. Differences were considered significant when p-value < 0.05.

FIGs. 3A-D are graphs showing the effects of the different diets on the development of glucose and insulin intolerance. Mice raised on the 4 diets were submitted to a glucose tolerance test (on week 9) and an insulin tolerance test (on week 10). For the glucose tolerance test (GTT, FIGs. 3A and 3B), mice were starved for 4h and injected (intra-peritoneal: ip) with glucose (1 g/kg). FIG. 3A: glycemia measured with a glucometer (AccuCheck Aviva™, Roche) was evaluated at -15, 0 (injection), 15, 30, 60, 90 and 120 min. FIG. 3B: AUC of the GTT results depicted in FIG. 3A. For the insulin tolerance tests (ITT, FIGs. 3C and 3D), mice were starved for 4h and injected ip with insulin (0,75USP/kg). FIG. 3C: Glycemia measured with a glucometer (AccuCheck Aviva™, Roche) was evaluated at 0 (injection), 15, 30, 45 and 60 min. post- injection. FIG. 3D: AUC of the ITT results depicted in FIG. 3C. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post- hoc Tukey tests. Differences were considered significant when p-value < 0.05.

FIG. 3E is a graph showing the development of glucose intolerance in mice fed on HFD to induce obesity. At the first day of obesity induction and 8 weeks later (weeks 5 and 13 of life, respectively), mice were starved for 6h and injected (ip) with glucose (1 g/kg). Glycemia measured with a glucometer (AccuCheck™ Aviva, Roche) was evaluated at -15, 0 (injection), 15, 30, 60, 90 and 120 min.

FIGs. 3F-3M are graphs showing the effect of the different diets on glucose intolerance in obese mice. Mice were submitted to a GTT towards the end of the obesity induction and after 8 weeks on the diet protocol (at 13 and 23-week old). FIGs. 3J-3M: AUC of the GTT data depicted in FIGs. 3F-3I, respectively. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post-hoc Tukey tests. Differences were considered significant when p-value < 0.05. FIGs. 3N-3U are graphs showing the effect of the different diets on insulin tolerance in obese mice. Mice were submitted to an ITT towards the end of the obesity induction and after 9 weeks on the diet protocol (at 14 and 24-week old). FIGs. 3R-3U: AUC of the GTT data depicted in FIGs. 3N-Q, respectively. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post-hoc Tukey tests. Differences were considered significant when p-value < 0.05.

FIGs. 4A-C are graphs depicting the effects of the different diets on the liver phenotype. Livers were dissected at the end of the 10-week diets. FIG. 4A (Liver appearance): Photographs of representative dissected livers for each group (n=8 per group). FIG. 4B (Liver weight): Liver weight averages for each group (n=8 per group). FIG. 4C (Liver triglyceride content): Liver triglyceride content was quantified using a colorimetric kit (Cayman™, Cat. #0010303) according to the manufacturer's instructions. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post- hoc Tukey tests. Differences were considered significant when p-value < 0.05.

FIGs. 4D and 4E are graphs depicting the liver weight of obese mice following treatment with the LFD, HFD, MCT-20 and MCT-40 diets. Livers were dissected at the end of the feeding protocol (week 25). FIG. 4D: Liver weight averages for each group (n=8 per group). FIG. 4E: Liver weight was divided by body mass (measured at week 24) for each mice. Data is presented as average ± standard error. Results from the 4 groups were compared using a one- way ANOVA with post-hoc Tukey tests. Differences were considered significant when p-value < 0.05.

FIGs. 5A and 5B show the effects of the different diets on the basal phosphorylation level of hepatic AKT, a key protein kinase in insulin signal transduction. Hepatic proteins were extracted from the dissected livers. In brief, liver samples (100-120 mg) were homogenized with a tissue grinder (Dounce homogenizer) in 1 ml of Radio-lmmunoprecipitation Assay cold buffer (RIPA, 50 mM Tris-HCI, 150 mM sodium chloride, 1 % NP-40, 0.5% sodium deoxycholate, 0.1 % sodium dodecyl sulfate, 200 μΜ sodium orthovanadate, 1 mM phenylmethylsulfonyl fluoride, ^g/mL leupeptin, 10 μg/mL aprotinin, pH 7.8). Following a centrifugation at 1 1 OOOg, for 15 min. at 4 °C, the clarified supernatant was isolated and protein concentration determined in this supernatant by Bradford protein assay. Phosphorylation levels of AKT (Ser473) were evaluated by Western blot (In brief, 20-100 μg of protein was mixed with 4 μΙ of 3X Laemmli sample buffer (2% SDS, 2% β-mercaptoethanol, 10% v/v glycerol and 50 mg/L bromophenol blue in 0.1 M Tris-HCI buffer, pH 6.8), heated at 100 °C for 5 min, subjected to SDS-PAGE and then transferred to Immobilon-P membranes for immunoblotting. Membranes were incubated for 1 h in blocking buffer (19 TBS, 0,1 % Tween™-20: TBST) containing 5% milk and then overnight in 1X TBST containing 5% milk and 1 :1000 of anti-p-AKT (Ser473) and anti-cyclophiline-B primary antibodies. Then, membranes were incubated for 1 h at room temperature in 1X TBST in the presence of 1 :2000 of anti-rabbit secondary antibody linked to the horseradish peroxidase, and the exposed to ECL reagent to reveal signals) and quantified by densitometry relative to Cyclophilin B, a housekeeping protein. Data is presented as average ± standard error. Results from the 4 groups were compared using a one-way ANOVA with post-hoc Tukey tests. Differences were considered significant when p-value < 0.05. FIG. 5A: Results in normal mice fed with the different diets for 10 weeks. FIG. 5B: Results in obese mice, i.e. fed on HFD for 10 weeks to induce obesity prior to LFD, HFD, MCT-20 and MCT-40 diets.

DISCLOSURE OF INVENTION

The use of the terms "a" and "an" and "the" and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to") unless otherwise noted.

Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All subsets of values within the ranges are also incorporated into the specification as if they were individually recited herein.

All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed.

Herein, the term "about" has its ordinary meaning. The term "about" is used to indicate that a value includes an inherent variation of error for the device or the method being employed to determine the value, or encompass values close to the recited values, for example within 10% or 5% of the recited values (or range of values).

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

In the studies described herein, the present inventors have shown in animal models that administration of a diet enriched in medium chain triglycerides (MCT) reduces or prevents several metabolic-related conditions, notably the increase in body weight, insulin resistance, glucose intolerance, and/or hepatic lipid deposition. The beneficial effects of MCT were obtained in normal mice administered with MCT prior to the development of the metabolic- related conditions, as well as in obese mice that had already developed one or more of these conditions.

Accordingly, in a first aspect, the present invention provides a method for preventing or reducing a metabolic condition (e.g., a pre-diabetic condition) in a subject, said method comprising administering a composition comprising an effective amount of MCT or medium- chain fatty acid (MCFA), to said subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, non-alcoholic fatty liver disease (NAFLD) or nonalcoholic hepatic steatosis (NASH).

In another aspect, the present invention provides the use of a composition comprising an effective amount of MCT or MCFA for preventing or reducing a metabolic condition (e.g., a pre-diabetic condition) in a subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, NAFLD or NASH.

In another aspect, the present invention provides the use of a composition comprising an effective amount of MCT or MCFA for the manufacture of a medicament for preventing or reducing a metabolic condition (e.g., a pre-diabetic condition) in a subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, NAFLD or NASH.

In another aspect, the present invention provides a composition comprising an effective amount of MCT or MCFA for preventing or reducing a metabolic condition (e.g., a pre-diabetic condition) in a subject, wherein said metabolic condition is one or more of insulin resistance, glucose intolerance, NAFLD or NASH.

In an embodiment, the composition is for preventing or reducing insulin resistance. In another embodiment, the composition is for preventing or reducing glucose intolerance. In another embodiment, the composition is for preventing or reducing NAFLD. In another embodiment, the composition is for preventing or reducing NASH.

In another aspect, the present invention provides a method for increasing the responsiveness or sensitivity of a subject to insulin stimulation, said method comprising administering a composition comprising an effective amount of MCT or MCFA, to said subject.

In another aspect, the present invention provides the use of a composition comprising an effective amount of MCT or MCFA for increasing the responsiveness or sensitivity of a subject to insulin stimulation.

The term "medium chain fatty acid (MCFA)" as used herein refers to a saturated fatty acid composed of 6 to 12 carbon atoms, preferably 6 to 10 carbon atoms. In an embodiment, the MCFA is composed of 8 to 10 carbon atoms. The MCFA may be in the form of triacyl- glycerol, diacyl-glycerol, or monoacyl-glycerol. In an embodiment, the MCFA is octanoic acid and/or decanoic acid.

In an embodiment, the MCFA may be in the form of a triglyceride (MCT). In particular, the MCFA may be in the form of a MCT in which each of the fatty acid moieties comprises the same number of carbons. For example, the MCT may comprise three decanoic (capric) acid moieties or three octanoic (caprylic) acid moieties.

The term "medium-chain triglycerides" or "MCTs" means any glycerol molecule ester- linked to three MCFA molecules. MCTs may be represented by the following formula:

H

i

H-C- OOCR' FTCOO-C— H

H-C- OOCFT

j

H

where R', R" and R'" are (saturated) fatty acids having 6-12 or 6-10 carbons in the carbon backbone esterified to the glycerol backbone. The lipid structure of this invention may be prepared by any process known in the art, such as direct esterification, rearrangement, fractionation, transesterification, or the like. For example, the lipids may be isolated from, or derived from, a source of vegetable oil, such as coconut oil, such as through a rearrangement process or the like. The length and distribution of the chain length may vary depending on the source oil. Typically, MCT oil contains 60% C8 and 40% C10. Such oils are commonly derived from palm kernel and coconut oils. MCTs containing greater than about 95% C8 at R', R" and R'" can be made by semi-synthetic esterification of octanoic acid to glycerin. Also useful herein are mixtures comprising MCT or MCFA with about 50% total C8 and/or about 50% total C10. Commercial sources for the foregoing MCT or MCFA compositions are available and known to the skilled artisan. Because MCT are metabolized into MCFA, which are readily oxidized, the administration of MCFA have the same effect as the administration of MCT themselves. The composition may comprise a single MCT (a MCT comprising a single type of MCFA, e.g., capric acid) a single MCFA, or any mixture of different MCT or MCFA, in any proportion. In an embodiment, the composition comprises a mixture of at least two, three or four different MCT or MCFA. In an embodiment, the composition comprises a mixture of two different MCT or MCFA, for example a MCT comprising capric acids and a MCT comprising caprylic acids, or a MCFA mixture comprising capric acids and caprylic acids. The composition may comprise an approximately equal ratio of caprylic acids and capric acids, or a ratio ranging from 10:1 to 1 :10, e.g., 10:1 , 8:1 , 6:1 , 4:1 , 2:1 , 1 :2, 1 :4, 1 :6, 1 :8 or 1 :10 wt/wt.

The composition may be a food composition, wherein the MCT or MCFA are inserted or mixed into a food substance. The term "food composition" includes any food, feed, snack, food supplement, treat, meal substitute, or meal replacement, whether intended for a human or an animal. In an embodiment, the food composition or food supplement is intended for human. The composition may be in the form of a food stuff or supplement, for example a human food stuff. The composition may be in the form of a nutritional supplement. A nutritional supplement refers to a product which is intended to supplement the general diet of a subject, and which may be in any form, e.g., solid, liquid (e.g., oil), gel, tablets, capsules, powder, and the like. The composition may be in the form of a complete nutritional product. A complete nutritional product refers to a product which is intended to be the sole item or meal or diet consumed by a subject.

The composition may be in the form of a beverage, mayonnaise, margarine, low fat spread, a dairy product, a cheese spread, processed cheese, a dairy dessert, a flavoured milk, cream, a fermented milk product, cheese, butter, a condensed milk product, an ice cream mix, a soya product, pasteurised liquid egg, a bakery product, a confectionary product, confectionary bar, chocolate bar, high fat bar, liquid emulsion, spray dried powder, freeze dried powder, pudding, gel, jelly, yoghurt or a food with a fat-based or water-containing filling. In an embodiment, the composition is in the form of an oil, for example a concentrated MCT oil. The incorporation of MCT into food products such as baked goods, beverages, chewing gum, confections and frostings, dairy product analogues, fats and oils, frozen dairy desserts, processed fruits, snack foods, adult nutritionals, cheeses and cheese spreads and soft candies is recognized as safe by the FDA (GRAS Notice No. 449).

For pet foods and food products formulated for human consumption, the amount of

MCT or MCFA as a percentage of the composition can be in the range of about 1 % to about 20% of the composition, although a lesser or greater percentage can be supplied. In various embodiments, the amount can be about or between any of the following: 1 .0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 1 1 %, 1 1 .5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 20% or more, of the composition. Dietary supplements may be formulated to contain several fold higher concentrations of MCT, to be amenable for administration to an animal in the form of a tablet, capsule, liquid concentrate, or other similar dosage form, or to be diluted before administration, such as by dilution in water, spraying or sprinkling onto a pet food, and other similar modes of administration. For a dietary supplement, MCT or MCFA alone may be administered directly to the subject or applied directly to the subject's regular food. Dietary supplement formulations in various embodiments contain about 30% to about 100% MCT or MCFA, although lesser amounts may also be used.

The composition may comprise a concentration of about 5 g/L to 150 g/L of MCT and/or MCFA. The concentration may be about 5 g/L, 10 g/L, 15 g/L, 20 g/L, 30 g/L, 40 g/L, 50 g/L, 60 g/L, 70 g/L, 80 g/L, 90 g/L, 100 g/L, 1 10 g/L, 120 g/L, 130 g/L, 140 g/L, 150 g/L, 175 g/L, 200 g/L, 225 g/L or 250 g/L of MCT and/or MCFA. In general terms, administration of the composition may be by an oral route or another route into the gastro-intestinal tract, for example the administration may also be by gastric tube feeding.

The composition may be enriched with MCT or MCFA. In particular, the composition may be enriched with capric acids and caprylic acids, or MCT comprising capric acids and caprylic acids.

"Enriched" means that the MCT or MCFA has been added to the composition. For example, the MCT or MCFA may be spiked (i.e. added within or into) the composition.

In one embodiment, where a food or food extract natively contains MCT or MCFA, enriched with a MCT or MCFA means that the enriched food or food extract comprises a greater amount of the MCT or MCFA than occurs natively in the food or food extract.

For example, an enriched composition, food or food extract may comprise at least 1.5-, at least 2-, at least 5-, at least 10-, at least 20-, at least 50- or at least 100-fold more MCT or MCFA than an equivalent native composition, food or food extract which has not been enriched.

The compositions can optionally comprise one or more supplementary substances that promote or sustain general health, e.g., nutrients (vitamins, minerals), proteins, fiber sources, prebiotics or probiotics, etc. In an embodiment, the composition is free of a beta- hydroxybutyrate (BHB) compound, e.g., beta-hydroxybutyrate or a salt thereof. In another embodiment, the methods/uses described herein do not comprise the use/administration of a combination of MCT or MCFA with a beta-hydroxybutyrate compound.

The skilled artisan will also appreciate that in formulating the food compositions described herein, the formulation may vary slightly, so as to allow consideration by the formulator of the price and/or availability of certain ingredients in the compositions, as well as the batch-to-batch variation in the analysis of certain ingredients. Thus, a given food composition or formulation may vary slightly from batch to batch, plant to plant, or even season to season depending on such factors. Notwithstanding such variation in specific ingredients selected for manufacturing a particular batch of a food composition, the overall composition (for example, analysis of protein, carbohydrate, fat, fiber, or other component) may be held constant or at least substantially constant, for example, in accordance with a label claim, such as a claim or guarantee of a minimum or maximum percent of a particular component.

The composition may alternatively be in the form of a tablet, dragees, capsule, gel cap, powder, granule, solution, emulsion, suspension, coated particle, spray-dried particle or pill. The composition may be usable for reconstitution in water. The composition may be in the form of a powder, e.g., a spray-dried powder or a freeze-dried powder. The composition may be an oil-in-water emulsion. In embodiments, the composition comprises one or more pharmaceutically acceptable carrier(s) or excipient(s). Such compositions may be prepared in a manner well known in the pharmaceutical art. In an embodiment, the composition is for oral administration.

An "excipient" or "carrier" as used herein, has its normal meaning in the art and is any ingredient that is not an active ingredient (i.e., MCT or MCFA) itself. Excipients include for example binders, lubricants, diluents, fillers, thickening agents, disintegrants, plasticizers, coatings, barrier layer formulations, lubricants, stabilizing agent, release-delaying agents and other components. "Pharmaceutically acceptable excipient" as used herein refers to any excipient that does not interfere with effectiveness of the biological activity of the active ingredients (i.e., MCT or MCFA) and that is not toxic to the subject, i.e., is a type of excipient and/or is for use in an amount which is not toxic to the subject. Excipients are well known in the art, and the present system is not limited in these respects. Examples of excipients include binders (binding agents), thickening agents, surfactants, diluents, release-delaying agents, colorants, flavoring agents, fillers, disintegrants/dissolution promoting agents, lubricants, plasticizers, silica flow conditioners, glidants, anti-caking agents, anti-tacking agents, stabilizing agents, anti-static agents, swelling agents and any combinations thereof. As those of skill would recognize, a single excipient can fulfill more than two functions at once, e.g., can act as both a binding agent and a thickening agent. As those of skill will also recognize, these terms are not necessarily mutually exclusive. The preparation of pharmaceutical compositions is described, for example, in Remington: The Science and Practice of Pharmacy, by Loyd V Allen, Jr, 2012, 22^nd edition, Pharmaceutical Press; and Handbook of Pharmaceutical Excipients, by Rowe et al., 2012, 7^th edition, Pharmaceutical Press.

Any suitable amount of the pharmaceutical composition may be administered to a subject. The dosages will depend on many factors including the mode of administration. Typically, the amount of MCT or MCFA contained within a single dose will be an amount that effectively prevents or reduces a metabolic condition as defined herein without inducing significant toxicity.

For the prevention or reduction in the severity of a given disease or condition, the appropriate dosage of the agent (MCT or MCFA), or composition comprising same, will depend on the type of disease or condition to be treated, the severity and course of the disease or condition, whether the agent/composition is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the agent/composition, and the discretion of the attending health professional (e.g., physician or nutritionist). The agent/composition is suitably administered to the patient at one time or over a series of treatments. Preferably, it is desirable to determine the dose-response curve in vitro, and then in useful animal models prior to testing in humans.

The skilled artisan will understand how to determine the appropriate amount of MCT or MCFA, and any other ingredients to be added to a given composition. The skilled formulator may consider important the animal's species, age, size, weight, health, and the like in determining how best to formulate a particular composition, food, or pharmaceutical composition comprising the MCTs and other components. Other factors that may be taken into account for formulation include the type of composition (e.g. , pet food composition versus dietary supplement), the desired dosage of each component (MCT or MCFA), the average consumption of specific types of compositions by different animals (e.g., based on species, body weight, activity/energy demands, and the like) and the manufacturing conditions under which the composition is prepared. In one embodiment, the concentrations of MCT or MCFA and other ingredients to be added to the composition are calculated on the basis of the energy and nutrient requirements of the animal. When formulating the compositions of the present invention, a skilled can determine the amounts of the MCT or MCFA and other components of the compositions and of other compounds or ingredients, in for example a food composition, based upon the desired dosages and the characteristics of the animal.

In an embodiment, the daily dose of MCT or MCFA can be from about 0.01 g/kg to about 10 g/kg body weight (BW) of the animal. In an embodiment, the daily dose of MCT or MCFA can be from about 0.05 g/kg to about 1 g/kg BW of the animal, or from about 0.1 g/kg to about 0.5 g/kg BW of the animal.

The composition may be in a form suitable for delivering a dosage of MCT or MCFA of at least about 1 g, 5 g, 10 g, 15 g, 20 g, 30 g, 40 g, 50 g, 75 g or 100 g per day. The composition may be in a form suitable for delivering a dosage of at least about 5 g to about 500, 400, 300, 200 or 150 g MCT or MCFA per day. In embodiments, the dosage may be about 5 g, 10 g, 15 g, 20 g, 30 g, 40 g, 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 1 10 g, 120 g, 130 g, 140 g, 150 g, 175 g, 200 g, 225 g, 250 g or 500 g MCT or MCFA per day. In further embodiments, the dosage may be about 5 g to about 20 g / day.

In an embodiment, the amount administered daily corresponds to an amount of at least about 0.1 g in mice. In an embodiment, the amount administered daily corresponds to an amount of at least about 0.2 g in mice. In an embodiment, the amount administered daily corresponds to an amount of at least about 0.3 g in mice. In an embodiment, the amount administered daily corresponds to an amount of at least about 0.35 g in mice. In an embodiment, the amount administered daily to the animal corresponds to an amount of at least about 0.4 g in mice. In an embodiment, the amount administered daily corresponds to an amount of at least about 0.5 g in mice. In an embodiment, the amount administered daily corresponds to an amount of at least about 0.6 g in mice. In an embodiment, the amount administered daily corresponds to an amount of about 0.1 to about 1 g in mice. In an embodiment, the amount administered daily corresponds to an amount of about 0.2 to about 0.8 g in mice. In an embodiment, the amount administered daily corresponds to an amount of about 0.3 to about 0.6 g in mice. Methods to convert animal dose to human equivalent doses (HED) are known in the art (see, e.g., Nair and Jacob, J Basic Clin Pharm. March 2016-May 2016; 7(2): 27-31 ; Rockville, MD: US Food and Drug Administration; 2005. USFDA. Guidance for Industry: Estimating the Maximum Safe Starting Dose in Adult Healthy Volunteer). For example, to convert a dose in mg/kg in mice to HED, the mice dose should be divided by about 12.3.

The composition described herein may be administered for any suitable period of time for mediating the desired effect, for example for at least 1 week, at least two weeks, at least one month, at least 3 months or at least 6 months.

The composition described herein may be administered on an as-needed or as-desired basis of varying or regular frequency. A goal of regular ingestion is to provide the animal with a regular and consistent dose of the composition or the direct or indirect metabolites that result from such ingestion. Such regular and consistent dosing will tend to create constant blood levels of the components of the compositions or their direct or indirect metabolites. Thus, regular administration can be once monthly, once weekly, once daily, or more than once daily. Similarly, administration can be every other day, week, or month, every third day, week, or month, every fourth day, week, or month, and the like. Administration can be multiple times per day. When utilized as a supplement to ordinary dietetic requirements, the composition may be administered directly to the animal, e.g., orally, or otherwise. The compositions can alternatively be contacted with, or admixed with, daily feed or food, including a fluid, such as drinking water, or an intravenous connection for an animal that is receiving such treatment. When utilized as a daily feed or food, administration will be well known to those of ordinary skill.

These are simply guidelines since the actual dose must be carefully selected and titrated by the attending health professional (e.g., physician or nutritionist) based upon clinical factors unique to each patient or by a nutritionist. The optimal daily dose will be determined by methods known in the art and will be influenced by factors such as the age and/or weight of the patient and other clinically relevant factors. In addition, patients may be taking medications for other diseases or conditions. The other medications may be continued during the time that MCT or MCFA is given to the patient, but it is particularly advisable in such cases to begin with low doses to determine if adverse side effects are experienced.

The term "preventing" as used herein means to administer a composition comprising MCT or MCFA as described herein to a subject not showing any symptoms of a pre-diabetic condition, to reduce or prevent development of at least one symptom associated with the pre- diabetic condition, or to delay the onset of the pre-diabetic condition.

The term "reducing a metabolic condition" as used herein means to administer a composition comprising MCT or MCFA as described herein to a subject already showing one or more symptoms of the metabolic condition, to reduce the severity of the symptom(s) or to avoid or delay the progression or worsening of the metabolic condition, e.g. into diabetes. In an embodiment, the composition is used to prevent or delay the progression of a pre-diabetic condition (e.g., insulin resistance or glucose intolerance) into diabetes. In an embodiment, the composition is used to prevent or delay the progression of NAFLD to NASH.

In an embodiment, the metabolic condition is a pre-diabetic condition. In an embodiment, the metabolic condition comprises NAFLD. In an embodiment, the metabolic condition comprises NASH. In an embodiment, the metabolic condition comprises insulin resistance. In an embodiment, the metabolic condition comprises IGT. In an embodiment, the metabolic condition comprises NAFLD or NASH, insulin resistance and IGT.

The term "subject" as used herein means any animal that could benefit from one or more of the methods/uses described herein. Generally, the animal is a human, avian, bovine, canine, equine, feline, hircine, lupine, murine, ovine, and porcine animal. A "companion animal" is any domesticated animal, and includes, without limitation, cats, dogs, rabbits, guinea pigs, ferrets, hamsters, mice, gerbils, horses, cows, goats, sheep, donkeys, pigs, and the like. In one example, the animal can be a human or a companion animal such as a dog or cat.

In an embodiment, the subject is an adult human subject. In an embodiment, the subject is not a neonate, for example a premature baby. In an embodiment, the subject suffers from obesity or is at risk of suffering from obesity, e.g. is overweight. In an embodiment, the subject has a Body Mass Index (BMI) that is at least 25, at least about 26, at least about 27, at least about 28, at least about 29, or at least about 30.

MODE(S) FOR CARRYING OUT THE INVENTION

The present invention is illustrated in further details by the following non-limiting examples.

Example 1 : Effects of different diets on weight gain

Four groups of 6-week-old C57BL6/J mice (n=8 per group) were fed during 10 weeks with the diets described below.

The following isocaloric diets (similar caloric input per mass unit; kcal/g) were used

(see FIGs. 1A and 1 B):

• LFD (low fat diet); 10% energy (kcal) as long chain triglycerides (LCT); negative control of obesity induction. The LFD is a rodent diet with 10 kcal% fat (Research Diets, Inc., Cat. # D12450H, listed standard diet).

· HFD (high fat diet); 45% energy (kcal) as LCT; positive control of obesity induction. The HFD is a rodent Diet with 45 kcal% fat (Research Diets, Inc., Cat. # D12451 , listed standard diet).

• MCT-20, high fat diet in which a portion of LCT was replaced by MCT; 20% energy (kcal) as MCT and 25% as LCT. MCT-20 (D08041702, rodent diet with 45 kcal% fat, non listed diet, customized by Research Diets, Inc.

· MCT-40, high fat diet in which a portion of the LCT was replaced by MCT; 40% energy (kcal) as MCT and 5% as LCT. MCT-40 (D08041701 , rodent diet with 45 kcal% fat, non listed diet, customized by Research Diets, Inc. The MCT source for MCT-20 and MCT-40 is MCT oil from ALZO international Inc. comprising 60/40 (wt/wt) Octanoic/Decanoic acid triglycerides.

The detailed composition of the different diets is depicted in FIG. 1A.

It was estimated that each mouse ate about 3 g of food per day. Based on the proportion of MCT in the MCT-20 and MCT-40 diets (about 10% and 20%, respectively - see Table 1), the daily consumption of MCT by the mice of the MCT-20 and MCT-40 groups was estimated to about 0.3 g and 0.6 g, respectively.

The data presented in FIG. 2A shows that the total energy consumption (i.e. the cumulative energy consumed during the 10-week diet period) was similar for the different groups. However, replacement of a portion of the LCT by MCT prevented in a dose-dependent manner the weight gain associated with the HFD diet, as shown in FIG. 2B.

To assess the curative effect of MCT, 32 C57BL6/J mice (5-week-old) were first fed with a HFD for 10 weeks to induce obesity. The obese mice were then split into 4 groups of 8 mice, which were fed for an additional 10 weeks with the four diets described above (LFD, HFD, MCT-20 and MCT-40) (FIG. 1C). As shown in FIG. 2C, an important weight gain was observed after 10 weeks of HFD. The data presented in FIG. 2D and FIG. 2E show that the total food uptake and energy consumption was similar for the different groups. FIGs. 2F-2H show that the MCT-20 diet reduces the food-induced weight gain observed with HFD, and the MCT-40 diet led to a significant weight loss relative to HFD, similar to the LFD. Thus, these results provide evidence that MCT may be useful for limiting or reducing weight gain.

Example 2: Effects of the different diets on glucose and insulin tolerance

Normal mice fed on the four diets described above were submitted to a glucose tolerance test (on week 9) and an insulin tolerance test (on week 10). The results depicted in FIGs. 3A and 3B show that the presence of MCT in the HFD diet prevented the development of glucose intolerance in the mice. High levels of MCT also prevented the induction of insulin resistance in the mice, as shown in FIGs. 3C and 3D.

The curative effect of MCT on glucose and insulin tolerance in obese mice was also assessed. As shown in FIG. 3E, 8 weeks of HFD led to glucose intolerance. However, long-term feeding of obese glucose-intolerant mice with MCT-40 rescues glucose tolerance at a higher extent than the control LFD (FIGs. 3F-3M). Also, long-term feeding of obese glucose-intolerant mice with MCT-40 significantly rescues insulin sensitivity at a similar extent as the control LFD, and MCT-20 stabilizes the state of insulin insensitivity in obese mice (FIGs. 3N-3U).

These results thus provide evidence that MCT may be useful for preventing the apparition of pre-diabetic symptoms/conditions like insulin resistance and glucose intolerance, as well as for managing these symptoms/conditions in obese subjects. Example 3: Effects of the different diets on the liver

The livers of mice fed on the four diets described above were dissected and analyzed at the end of the 10-week period. Analysis of liver appearance (FIG. 4A) revealed the accumulation of fat in the livers of mice fed with HFD, and effect that was prevented in the presence of MCT in the diets. The presence of MCT in the diets was also associated with a significant reduction in liver weight with the MCT-40 diet (FIG. 4B), as well as with a clear trend toward decreased levels of liver triglycerides (TG) (FIG. 4C).

The effect of MCT on liver weight in obese mice was also assessed. As shown in FIGs. 4D and 4E, feeding of obese mice with MCT-40 led to a significant reduction in liver weight.

These results thus provide evidence that MCT may be useful for reducing lipid accumulation in the liver, and thus for preventing the development of and/or managing fatty liver disease/liver steatosis.

The effects of the diets on the phosphorylation status of hepatic AKT (Ser473) was next assessed. The PI3K/AKT signaling pathway is involved in the intracellular transduction of the insulin signal, and the levels of basal hepatic Ser473 AKT phosphorylation is indicative of the responsiveness of this pathway to insulin stimulation. As shown in FIG. 5A, the MCT-20 and MCT-40 diets significantly raised the basal phosphorylation level of hepatic AKT at Ser473, and the MCT-40 diet also led to a rescue of the basal phosphorylation level of hepatic AKT in obese mice (FIG. 5B). These results could explain, at least in part, the ability of the MCT-containing diets to prevent the induction of insulin resistance and/or glucose intolerance, and to manage insulin resistance and/or glucose intolerance in obese mice, shown in Example 2.

Although the present invention has been described hereinabove by way of specific embodiments thereof, it can be modified, without departing from the spirit and nature of the subject invention as defined in the appended claims.

Claims

WHAT IS CLAIMED IS:

2. The method of claim 1 , wherein said metabolic condition is insulin resistance.

3. The method of claim 1 or 2, wherein said metabolic condition is glucose intolerance.

4. The method of any one of claims 1 to 3, wherein said metabolic condition is non- alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

5. The method of any one of claims 1 to 4, wherein said composition comprises at least about 5% of MCT or MCFA (wt/wt of the composition).

6. The method of any one of claims 1 to 4, wherein said composition comprises at least about 10% of MCT or MCFA (wt/wt of the composition).

7. The method of any one of claims 1 to 4, wherein said composition comprises about 5% to about 50% of MCT or MCFA (wt/wt of the composition).

8. The method of any one of claims 1 to 4, wherein said composition comprises about 10% to about 30% of MCT or MCFA (wt/wt of the composition).

9. The method of any one of claims 1 to 8, wherein said composition is administered prior to or after the development of said metabolic condition.

10. The method of any one of claims 1 to 9, wherein said composition is administered at least once daily.

1 1. The method of any one of claims 1 to 10, wherein said MCT or MCFA comprise C8 fatty acids (caprylic acids).

12. The method of any one of claims 1 to 1 1 , wherein said MCT or MCFA comprise C10 fatty acids (capric acids).

13. The method of any one of claims 1 to 12, wherein said MCT or MCFA comprise (i) caprylic acids and (ii) capric acids.

14. The method of claim 13, wherein said MCT or MCFA comprise at least 40% of caprylic acids and at least 40% of capric acids.

15. The method of any one of claims 1 to 14, wherein said composition is a food composition or food supplement.

16. The method of any one of claims 1 to 15, wherein said composition is a concentrated MCT or MCFA oil composition.

17. The method of any one of claims 1 to 14, wherein said composition is a pharmaceutical composition.

20. The use of claim 18 or 19, wherein said metabolic condition is insulin resistance.

21. The use of claim 18 or 19, wherein said metabolic condition is glucose intolerance.

22. The use of any one of claims 18 to 21 , wherein said metabolic condition is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic hepatic steatosis (NASH).

23. The use of any one of claims 18 to 22, wherein said composition comprises at least about 5% of MCT or MCFA (wt/wt of the composition).

24. The use of any one of claims 18 to 22, wherein said composition comprises at least about 10% of MCT or MCFA (wt/wt of the composition).

25. The use of any one of claims 18 to 22, wherein said composition comprises about 5% to about 50% of MCT or MCFA (wt/wt of the composition).

26. The use of any one of claims 18 to 22, wherein said composition comprises about 10% to about 30% of MCT or MCFA (wt/wt of the composition).

27. The use of any one of claims 18 to 26, wherein said composition is used prior to or after the development of said metabolic condition.

28. The use of any one of claims 18 to 27, wherein said composition is administered at least once daily.

29. The use of any one of claims 18 to 28, wherein said MCT or MCFA comprise C8 fatty acids (caprylic acids).

30. The use of any one of claims 18 to 29, wherein said MCT or MCFA comprise C10 fatty acids (capric acids).

31. The use of any one of claims 18 to 30, wherein said MCT or MCFA comprise (i) caprylic acids and (ii) capric acids.

32. The use of claim 31 , wherein said MCT or MCFA comprise at least 40% of caprylic acids and at least 40% of capric acids.

33. The use of any one of claims 18 to 32, wherein said composition is a food composition or food supplement.

34. The use of any one of claims 18 to 33, wherein said composition is a concentrated MCT or MCFA oil composition.

35. The use of any one of claims 18 to 32, wherein said composition is a pharmaceutical composition.