WO2012060718A1

WO2012060718A1 - Preventing or treating metabolic syndrome by administering a trans fatty acid, or a salt, ester or precursor thereof, or sialic acid in free or bound form

Info

Publication number: WO2012060718A1
Application number: PCT/NZ2010/000240
Authority: WO
Inventors: Sally Diana Poppitt; Geoffrey Wayne Krissansen; Anne-Thea Mcgill; Neill Ward Haggarty; Alastair Kenneth Hugh Macgibbon; Paul Frank Davis
Original assignee: Sally Diana Poppitt; Geoffrey Wayne Krissansen; Anne-Thea Mcgill; Neill Ward Haggarty; Alastair Kenneth Hugh Macgibbon; Paul Frank Davis
Priority date: 2010-11-05
Filing date: 2010-12-01
Publication date: 2012-05-10

Abstract

The present invention relates to use of trans fatty acid or a salt, ester or precursor thereof or trans fatty acid-enriched milk fat comprising milk fat enriched with trans fatty acid or a salt, ester or precursor thereof; or sialic acid for preventing or treating conditions associated with metabolic syndrome such as fatty liver disease. The invention also relates to a pharmaceutical composition comprising CLA-enriched milk fat or sialic acid.

Description

PREVENTING OR TREATING METABOLIC SYNDROME BY ADMINISTERING A TRANS FATTY ACID, OR A SALT, ESTER OR PRECURSOR THEREOF, OR SIALIC

ACID IN FREE OR BOUND FORM.

TECHNICAL FIELD

The present invention relates to use of trans fatty acids or sialic acid to treat or prevent conditions associated with metabolic syndrome, for example, fatty liver disease. The invention also relates to a conjugated linoleic acid-enriched milk fat composition and its use in methods of treating or preventing conditions associated with metabolic syndrome or fatty liver disease.

BACKGROUND OF THE INVENTION

There is a worldwide epidemic of weight gain and obesity, with the World Health

Organisation estimating that more than 1 billion people globally are now overweight, and of these, 300 million are obese. This epidemic represents a major risk for the development of diseases such as diabetes, heart disease and some cancers. One of the adverse effects of weight gain is the increase in metabolic dysregulation, which leads to many of these diseases.

Metabolic syndrome is defined as a cluster of metabolic disorders that increase the risk of developing cardiovascular disease and type 2 diabetes mellitus. This cluster of metabolic disorders is characterised by central adiposity, adverse lipid and glucose profiles and hypertension. Other metabolic abnormalities that can develop include an increase in the pro-inflammatory profile, disordered blood clotting and 'lipid overspill' from adipose tissue into critical organs such as the liver, heart and skeletal muscle.

Non alcoholic fatty liver disease or non alcoholic steatohepatitis are highly prevalent diseases that are reported to accompany obesity and metabolic syndrome and can lead to cirrhosis and liver failure. Cardiovascular disease is a major consequence of obesity and metabolic syndrome and is currently/one of the leading causes of mortality in the developed world. It is thought to account for approximately 12 million deaths per year. Risk factors contributing to the development of cardiovascular disease include obesity, blood pressure and an individual's lipid profile. Lifestyle changes that make positive alterations to an individual's metabolic profile have been shown to consistently improve cardiovascular risk and it is thought dietary changes are a major driver underpinning the control of these metabolic abnormalities.

Current treatment or prevention strategies for reducing the risk of developing metabolic syndrome and its associated diseases are limited. Furthermore there are no current therapies for fatty liver disease, a focus of this invention, and obesity is commonly leading to liver transplant as there are no effective therapies for this extremely serious medical issue.

It is an object of the present invention to provide an improved or alternative means for treating or preventing metabolic syndrome conditions such as fatty liver disease or to at least provide the public with a useful choice. SUMMARY OF THE INVENTION

Accordingly, in a first aspect the present invention relates to a method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of a trans fatty acid, or a salt, ester or precursor thereof.

In one embodiment the trans fatty acid is present in a composition enriched in trans fatty acid, including a composition comprising one or more lipids enriched in trans fatty acid.

In one embodiment the trans fatty acid is conjugated linoleic acid (CLA).

Accordingly, the present invention relates to a method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of CLA, or a salt, ester or precursor thereof.

In one embodiment the conjugated linoleic acid is selected from one or more CLA isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof, including the cis-9,cis-l l, cis-9,trans-l l , trans-9,cis-l l , trans-9,trans-l l, cis- 10,cis-12, cis-10,tl2, trans-10,cis-12, and trans-10, trans-12 isomers, preferably the cis-9,cis-l l, cis- 9,trans-l l, trans-10,cis-12, or cis-10,cis-12 isomers.

In one embodiment the conjugated linoleic acid is ds-9, transA 1 CLA.

Accordingly, the present invention relates to a method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of ds-9, transA 1 CLA, or a salt, ester or precursor thereof.

In one embodiment the trans fatty acid is a precursor to CLA, such as vaccenic acid.

In one embodiment the method comprises administration of ds-9, transA 1 CLA and vaccenic acid.

In one embodiment the trans fatty acid is trans 16:1 , trans 18:1 fatty acid, or a combination thereof.

In another aspect the present invention relates to a method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of a trans fatty acid, or a salt, ester or precursor thereof.

In one embodiment the trans fatty acid is CLA.

Accordingly, the present invention relates to a method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of CLA, or a salt, ester or precursor thereof.

In one embodiment the conjugated linoleic acid is selected from one or more CLA isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof, including the cis-9,cis-l 1, cis-9,trans-l l, trans-9,cis-l l , trans-9,trans-l l, cis- 10,cis-12, cis-10,tl2, trans-10,cis-12, and trans- 10,trans- 12 isomers, preferably the cis-9,cis-l l, cis- 9,trans-l l, trans-10,cis-12, or cis-10,cis-12 isomers.

In one embodiment the conjugated linoleic acid is as-9, transA 1 CLA.

Accordingly, the present invention relates to a method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of is-

9, transA 1 CLA, or a salt, ester or precursor thereof.

In one embodiment the method comprises administration of as-9, transA 1 CLA and vaccenic acid.

In another aspect the present invention relates to a method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an ^• effective amount of milk fat enriched with trans fatty acid.

In one embodiment the trans fatty acid is CLA.

In one embodiment the milk fat is present in a composition, preferably a composition comprising CLA or a salt, ester, or precursor thereof.

In one embodiment the conjugated linoleic acid is selected from one or more CLA isomers of 9,11 -octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof, including the cis-9,cis-l l , cis-9,trans-l l, trans-9,cis-l l, trans-9,trans-l 1, cis-

10, cis-12, cis-10,tl 2, trans- 10,cis- 12, and trans- 10,trans- 12 isomers, preferably the cis-9,cis-l 1, cis- 9,trans-l l , trans-10,cis-12, or cis-10,cis-12 isomers.

In one embodiment the conjugated linoleic acid is trans-W CLA.

In one embodiment the milk fat or the composition is enriched with. both ds-9, trans-W CLA and vaccenic acid.

In one embodiment the trans fatty acid present in the milk fat or in the composition comprises, consists essentially of, or consists of vaccenic acid.

In one embodiment the trans fatty acid present in the milk fat or in the composition comprises, consists essentially of, or consists of as-9, trans-W CLA.

In one embodiment the milk fat or the composition is substantially free of the transAO, sA2 CLA isomer.

In one embodiment the CLA-enriched milk fat comprises at least about 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50% by weight of as-9, trans-W CLA or a salt, ester or precursor thereof and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to about 7%). Preferably the milk fat comprises at least about 2% as-9, transA 1 CLA by weight, preferably about 2 to 10% ds-9, trans-W CLA by weight, more preferably about 4 to 7% ds-9, trans-W CLA by weight and most preferably about 5% ds-9, trans-W CLA by weight.

In one embodiment the milk fat comprises trans fatty acids which comprise at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% ds-9, trans-W CLA or a salt, ester or precursor thereof by weight and useful ranges may be selected between any of these forgoing values (e.g. from about 80% to about 95%). Preferably the milk fat includes trans fatty acids comprising at least about 50% ds-9, trans-W CLA or a salt, ester or precursor thereof by weight, preferably about 70 to 80% ds-9, trans-W CLA or a salt, ester or precursor thereof by weight.

In one embodiment the milk fat comprises CLA isomers which comprise at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% ds-9, trans-W CLA by weight or a salt, ester or precursor thereof and useful ranges may be selected between any of these forgoing values (e.g. from about 80% to about 95%). Preferably the milk fat includes CLA isomers comprising at least about 50% ds-9, trans- 11 CLA by weight, preferably about 70 to 80% ds-9, trans-W CLA by weight.

In one embodiment the ds-9, transA 1 CLA is selected from ds-9, trans-W CLA derived from a natural source; synthetic ds-9, trans-W CLA; ds-9, trans-W CLA in free fatty acid form; ds-9, trans-W CLA bound to glycerol, a monoglyceride, a diglyceride or a triglyceride; ds-9, trans-X 1 CLA in esterified form; or mixtures thereof.

In one embodiment the milk fat is produced by enhancing natural levels of CLA in milk by feeding a milk producing mammal with a diet enriched in at least one fatty acid (e.g. linoleic acid).

In another embodiment the milk fat composition of the invention is prepared by combining a source of ds-9, trans-W CLA or a salt, ester or precursor thereof with milk fat.

In another aspect the present invention relates to a method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of a composition comprising sialic acid.

Another aspect of the invention relates to a method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of a composition comprising sialic acid.

In one embodiment the composition comprises bound sialic acid and free sialic acid.

In one embodiment the composition comprises sialic acid and at least one monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharide.

In one embodiment at least some of the sialic acid is bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharides.

In one embodiment at least some of the sialic acid is covalently bound to at least a portion of. the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or

oligosaccharides. In one embodiment the disaccharide is lactose.

In one embodiment the sialic acid is covalently bound to the lactose.

In one embodiment the composition comprises sialic acid and lactose, wherein the sialic acid is administered with lactose either simultaneously, separately or sequentially.

In one embodiment the composition additionally comprises minerals.

In one embodiment the minerals are selected from the group comprising calcium, potassium, sodium, phosphorus, magnesium or zinc.

In one embodiment the composition additionally comprises phospholipids.

In one embodiment the phospholipid is selected from the group comprising

phosphatidylethanolamine, phosphatidylinositol or sphingomyelin.

In one embodiment the composition comprises at least about 0.2. 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4-5 or 5% sialic acid and useful ranges may be selected between any of these forgoing values (e.g. from about 1% to about 4%). Preferably the composition comprises at least about 0.2% sialic acid, preferably about 1 to 2% sialic acid.

In one embodiment the composition comprises at least about 40, 45, 50, 55, 60, 65, 70, 75, 80,

85 or 90% lactose and useful ranges may be selected between any of these forgoing values (e.g. from about 42% to about 68%). Preferably the composition comprises at least about 50% lactose, preferably about 55 to 65% lactose.

In one embodiment the composition comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12% protein and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to 'about 10%). Preferably the composition comprises at least about 4% protem, preferably about 6 to 10% protein. '

In one embodiment the composition comprises 2, 4, 6, 7, 8, 10, 12, 14, 16, 18 or 20% minerals and useful ranges may be selected between any of these forgoing values (e.g. from about 8% to about 18%). Preferably the composition comprises at least about 6% minerals, preferably 10 to 18% minerals.

In one embodiment the composition comprises from about 40% to about 90% lactose and from about 0.2% to about 5% sialic acid. Preferably, the composition comprises from about 50% to about 70% lactose and from about 1% to about 3% sialic acid.

In one embodiment the composition comprises from about 70% to about 90% lactose and from about 0.2% to about 0.5% sialic acid.

In one embodiment the composition is Oligolac™.

In one embodiment the composition comprises from about 40% to about 80% lactose, from about 1% to about 5% sialic acid and from about 4% to about 18% minerals. In one embodiment the composition comprises from about 55% to about 65% lactose, from about 1% to about 2% sialic acid and from about 5% to about 18% minerals.

In one embodiment the composition is Sialyl Oligolac™.

The use of a trans fatty acid or a salt, ester or precursor thereof, in the preparation of a medicament for use in any of the methods described above, such as preventing or treating metabolic syndrome conditions in a subject, is also contemplated.

In one embodiment the trans fatty acid is a precursor to conjugated linoleic acid, such as vaccenic acid.

In one embodiment the method comprises administration of s-9, trans-W conjugated linoleic acid and vaccenic acid.

The use of a composition comprising sialic acid in the preparation of a medicament for use in any of the methods described above, such as preventing or treating metabolic syndrome condition in a subject, is also contemplated.

In one embodiment at least some of the sialic acid is bound to at least a portion of the one o more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharides.

In one embodiment at least some of the sialic acid is covalently bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or

oligosaccharides. ,

In one embodiment the disaccharide is lactose.

In one embodiment the sialic acid is covalently bound to the lactose.

In one embodiment the composition additionally comprises minerals.

In one embodiment the composition additionally comprises phospholipids.

In one embodiment the phospholipid is selected from the group comprising

phosphatidylethanolamine, phosphatidylinositol or sphingomyelin.

In one embodiment the composition comprises at least about 0.2, 0.5, 1 , 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5% sialic acid and useful ranges may be selected between any of these forgoing values (e.g. from about 1% to about 4%). Preferably the composition comprises at least about 0.5% sialic acid, preferably about 1 to 2% sialic acid. In one embodiment the composition comprises at least about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 or 90% lactose and useful ranges may be selected between any of these forgoing values (e.g. from about 42% to about 68%). Preferably the composition comprises at least about 50% lactose, preferably about 55 to 65% lactose.

In one embodiment the composition, comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, .10, 11 or

12% protein and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to about 10%). Preferably the composition comprises at least about 4% protein, preferably about 6 to 10% protein.

In one embodiment the composition is Oligolac™.

In one embodiment the composition comprises from about 40% to about 80% lactose, from about 0.5% to about 5% sialic acid and from about 4% to about 18% minerals.

In one embodiment the composition comprises from about 55% to about 65% lactose, from about 0.5% to about 2% sialic acid and from about 5% to about 18% minerals.

In one embodiment the composition is Sialyl Oligolac™.

A composition comprising a trans fatty acid or a salt, ester or precursor thereof, for use in any of the methods or treatments described above is also contemplated. For example, a composition comprising trans fatty acid for preventing or treating a metabolic syndrome condition in a subject in need thereof is specifically contemplated.

In one embodiment the method comprises administration of s-9, trans-X 1 CLA and vaccenic acid.

A composition comprising sialic acid for use in any of the methods or treatments described above is also contemplated. For example, a composition comprising sialic acid for^'preventing or treating a metabolic syndrome condition in a subject in need thereof is specifically contemplated.

In one embodiment the composition comprises bound sialic acid and free sialic acid. In one embodiment the composition comprises sialic acid and at least one monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharide.

In one embodiment at least some of the sialic acid is covalendy bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or

oligosaccharides.

In one embodiment the disaccharide is lactose.

In one embodiment the sialic acid is covalendy bound to the lactose. ;

In /one embodiment the composition additionally comprises minerals.

In one embodiment the composition additionally comprises phospholipids.

In one embodiment the phospholipid is selected from the group comprising

phosphatidylethanolamine, phosphatidylinositol or sphingomyelin.

In one embodiment the composition comprises at least about 0.2, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5% sialic acid and useful ranges may be selected between any of these forgoing values (e.g. from about 1% to about 4%). Preferably the composition comprises at least about 0.5% sialic acid, preferably about 1 to 2% sialic acid.

In one embodiment the composition comprises at least about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 or 90% lactose and useful ranges may be selected between any of these forgoing values (e.g. from about 42% to about 68%). Preferably the composition comprises at least about 50% lactose, preferably about 55 to 65% lactose.

In one embodiment the composition comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12% protein and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to about 10%). Preferably the composition comprises at least about 4% protein, preferably about 6 to 10% protein.

In one embodiment the composition comprises 2, 4, 6, 7, 8, 10, 12, 14, 16, 18 or 20% minerals and useful ranges may be selected between any of these forgoing values (e.g. from about 8% to about 18%). Preferably the composition comprises at least about 6% minerals, preferably 10 to 18% minerals. In one embodiment the composition comprises from about 40% to about 90% lactose and from about 0.2% to about 5% sialic acid. Preferably, the composition comprises from about 50% to about 70% lactose and from about 1% to about 3% sialic acid.

In one embodiment the composition comprises from about 70% to about 90% lactose and from about 0.2% to about 0.5% sialic acid. . ·

In one embodiment the composition is^' Oligolac™.

In one embodiment the composition is Sialyl Oligolac™.

Additionally, a composition comprising a trans fatty acid and one or more additional agents for use in any of the methods and treatments described above is also contemplated. For example, a composition comprising a trans fatty acid and one or more additional agents for improving the efficacy of the composition to prevent or treat a metabolic syndrome condition in a subject in need thereof is also contemplated.

In one embodiment the one or more additional agents are lactose and sialic acid.

Additionally, a composition comprising sialic acid and one or more additional agents for use in any of the methods and treatments described above is also contemplated. For example, a composition comprising sialic acid and one or more additional agents for improving the efficacy of the composition to prevent or treat metabolic syndrome conditions in a subject in need thereof is also contemplated.

In one embodiment the additional agent is a trans fatty acid, is-9, trans-X 1 CLA or vaccenic acid.

Accordingly, in one embodiment the composition comprises sialic acid and trans fatty acid, for example sialic acid (including free sialic acid, bound sialic acid, or a combination thereof) and CLA.

In one embodiment the additional agent is milk fat enriched with a trans fatty acid, s-9, trans- 11 -CLA or vaccenic acid.

In one embodiment the composition comprises sialic acid, lactose and one or more additional agents. ^v Accordingly, in one embodiment the composition comprises sialic acid and trans fatty acid, for example sialic acid (including free sialic acid, bound sialic acid, or a combination thereof) and CLA.

The following embodiments may relate to any of the above aspects.

In various embodiments the subject is suffering from or is susceptible to developing a metabolic syndrome condition.

In various embodiments the metabolic syndrome condition may be a condition selected from the group comprising central adiposity, obesity, dyslipidemia, cardiovascular disease, atherosclerosis, hypertension, Type-2 diabetes mellitus, hyperinsulinemia, insulin resistance, hyperglycaemia, nonalcoholic fatty liver disease, non-alcoholic steatohepatitis, microalbuminuria or hyperuricemia.

In various embodiments the metabolic syndrome condition may be characterised by one or more of the following: increased uric acid levels, increased triglyceride levels, increased postprandial lipemia, decreased high density lipoprotein cholesterol, decreased post-heparin lypolytic activity, increased sympathetic nervous system activity, increased sodium retention, increased C-reactive protein (CRP), increased plasminogen activator inhibitor (PAI-1), increased fibrinogen, or increased fasting glucose.

In various embodiments the composition of the invention further comprises one or more constituents (such as antioxidants) which prevent or reduce degradation of the composition during storage or after administration.

In various embodiments the composition is formulated as a food, drink, food additive, drink additive, dietary supplement, nutritional product, medicament, pharmaceutical or nutraceutical. Preferably, the composition is formulated as a powder, liquid, food bar, spread, sauce, ointment, tablet or capsule.

In various embodiments the composition is formulated for enteral administration, such as oral administration.

In various embodiments the composition of the invention is formulated as a pharmaceutical composition suitable for enteral administration, such as oral administration.

It is intended that reference to a range of numbers disclosed herein (for example, 1 to 10) also incorporates reference to all rational numbers within that range (for example, 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9 and 10) and also any range of rational numbers within that range (for example, 2 to 8, 1.5 to 5.5 and 3.1 to 4.7) and, therefore, all sub-ranges of all ranges expressly disclosed herein are hereby expressly disclosed. These are only examples of what is specifically intended and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application in a similar manner. In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents is not to be construed as an admission that such documents, or such sources of information, in any jurisdiction,, are prior art, or form part of the common general knowledge in the art. ^'.

Further aspects and advantages of the present invention will become apparent from the ensuing description which is given by way of example only.

DETAILED DESCRIPTION OF THE INVENTION

1. Definitions

The terms "CLA-enriched milk fat" and "milk fat enriched with c-9, t-11 CLA" are intended to mean milk fat that comprises a higher level of ds-9, transA 1 CLA or a salt, ester or precursor thereof than normal milk fat. Milk fat for use according to the invention may in one embodiment be sheep, goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human milk fat. Preferably the milk fat is bovine milk fat.

In one embodiment the CLA-enriched milk fat comprises at least about 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50% by weight of ds-9, transA 1 CLA or a salt, ester or precursor thereof and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to about 7%). Preferably the CLA-enriched milk fat comprises at least about 2% ds-9, trans-W CLA by weight, preferably about 2 to 10% ds-9, transA 1 CLA by weight, more preferably about 4 to 7% ds-9, trans-W CLA by weight and most preferably about 5% ds-9, trans-W CLA by weight.

In one embodiment the CLA-enriched milk fat comprises CLA isomers which comprise at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% by weight ds-9, trans- CLA or a salt, ester or precursor thereof and useful ranges may be selected between any of these forgoing values (e.g. from about 80% to about 95%). Preferably the CLA-enriched milk fat includes CLA isomers comprising at least about 50% ds-9, trans-W CLA by weight, preferably about 70 to 80% ds-9, trans- 11 CLA by weight.

In one embodiment, the ds-9, trans-W CLA isomer may be included in a composition of the invention in free fatty acid form. In another embodiment the CLA may be in an esterified form, including but not limited to methyl, ethyl and propyl esters. In another embodiment the CLA may be in a salt form, including but not limited to sodium salts and zinc salts. In a further embodiment, one or more ds-9, trans-W CLA molecules may be bound to a polyol such as glycerol or sphingosine, with or without other fatty acids, to form mono-, di- or tri-glycerides or phospholipids for example. In yet another embodiment, mixtures of these forms of ds-9, trans-W CLA may be included within a composition of the invention. In another embodiment a precursor of is-9, transA 1 CLA may be provided including but not limited to vaccenic acid (transA 1-octadecenoic acid).

Appropriate levels of the ds-9, transA X CLA isomer or a salt, ester or precursor thereof may be determined, obtained and provided by a skilled worker with regard to that skill and to the teaching of the present application. .

The term "conjugated linoleic acid" (CLA) means one or more CLA isomers selected from isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof, including the cis-9,cis-l l, cis-9,trans-l 1, trans-9,cis-l l , trans- 9,trans-l l, cis-10,cis-12, cis-10,trans-12, trans- 10,cis- 12, and trans-10,tl2 isomers, preferably the cis- 9,cis-l l, cis-9,trans-l l , trans-10,cis-12, and cis-10,cis-12 isomers.

The term "comprising" as used in this specification and the claims means "consisting at least in part of. When interpreting statements in this specification and the claims which include that term, the features, prefaced by that term in each statement or claim, all need to be present but other features can also be present. Related terms such as "comprise" and "comprises" are to be interpreted in the same manner.

The term "effective amount" is the amount required to confer therapeutic effect. The interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described by Freireich, et al. (1966). Body surface area can be approximately determined from height and weight of the subject. See, e.g., Scientific Tables, Geigy

Pharmaceuticals, Ardley, New York, 1970, 537. Effective doses also vary, as recognized by those skilled in the art, dependent on route of administration, excipient usage, and the like.

The term "oral administration" includes oral, buccal, enteral and intra-gastric administration. The term "sialic acid" as used in this specification means an N- or O-substituted derivative of neuraminic acid, including, for example N-acetylnemaminic acid.

The term "bound sialic acid" as used in this specification refers to sialic acid bound to another carbohydrate.

The term "Sialyl Oligolac™" as used in this specification means a composition derived from milk or colostrum comprising approximately 62% lactose, approximately 8% protein, approximately 4% water, approximately 0.1 % fat, approximately 19% ash and approximately 1.2% sialic acid. Approximately 88% of the sialic acid is bound to another carbohydrate. The remaining sialic acid is in the form of free sialic acid. The term "Oligolac™" when used alone refers to a less concentrated form of the same product that has not been subject to lactose reduction, and so comprises 85% lactose, approximately 0.3% sialic acid, with other components scaled accordingly.

A "subject" is an animal, preferably a mammal, more preferably a mammalian companion animal or human. Preferred companion animals include cats, dogs and horses. The terms "trans fatty acid-enriched milk fat" and "milk fat enriched with trans fatty acids" are intended to mean milk fat that comprises a higher level of trans fatty acids or a salt, ester thereof than normal milk. Milk fat for use according to the invention may in one embodiment be sheep, goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human milk fat.

Preferably the milk fat is bovine milk fat. . ^■ . _. ·

In one embodiment the trans fatty acid-enriched rnilk fat comprises at least about 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50% by weight of trans fatty acid or a salt, ester or precursor thereof and useful ranges may be selected between any of these forgoing values (e.g. from about 4% to about 7%). Preferably the trans fatty acid-enriched milk fat comprises at least about 6% trans fatty acid by weight, preferably about 6 to 35% trans fatty acid by weight, more preferably about 20 to 30% trans fatty acid by weight and most preferably about 25 to 28% trans fatty acid by weight. In one example, the trans fatty acid-enriched milk fat comprises about 6% ds-9, trans A 1 CLA by weight. Preferably the trans fatty acid-enriched milk fat comprises about 6% to about 35% ds-9, trans-X X CLA by weight. In another example, the trans fatty acid-enriched milk fat comprises at least about 6% vaccenic acid by weight. Preferably the trans fatty acid-enriched milk fat comprises about 6% to about 35% vaccenic acid by weight.

The term "treat" and its derivatives should be interpreted in their broadest possible context. The term should not be taken to imply that a subject is treated until total recovery. Accordingly, "treat" broadly includes maintaining a subject's disease progression or symptoms at a substantially static level, increasing a subject's rate of recovery, amelioration and/ or prevention of the onset of the symptoms or severity of a particular condition, or extending a patient's quality of life. The term "treat", also broadly includes the maintenance of good health for sensitive indmduals and building stamina for disease prevention.

2. Conjugated Linoleic Acid (CLA)

Conjugated Lmoleic Acid (CLA) represents a mixture of positional and geometric isomers of linoleic acid, an 18 carbon fatty acid comprising two double bonds which are conjugated. CLA is a natural but small component of fats derived from ruminant animals fed a conventional diet.

The two major jsoforms of CLA are ds-9, trans-X X CLA and trans-X O dsA 2 CLA. The ds-9, trans-X X CLA is the principal dietary form of CLA and accounts for in excess of 90% of the total CLA content in dairy products.

The ds-9, trans-X X CLA may be synthetic, derived from a natural source, or mixtures thereof. Natural sources of ds-9, trans-X X CLA are derived from ruminant animals, while synthetic CLA includes CLA that is chemically modified to improve potency, stability, transport and half-life. CLA derived from ruminants is the result of incomplete bio-hydrogenation of dietary unsaturated fatty acids in the rumen, or synthesis from vaccenic acid elsewhere within the animal's tissues. Sunflower and safflower seed oils, containing approximately 65% and 76% linoleic acid respectively, can also be used as raw material for CLA production. Optimal conditions used in commercial scale production results in approximately equal amounts of the isomers cis-9, trans-\ \ and transA Q, cisA2. A safflower based product can thus contain approximately 36% each of cis-9, trans-W and transAO, cisA2 isomers. Minor peaks include the cis, cis and trans, trans isomers of 9,11 and 10,12 CLA, each around 0.5 to 1%. Traces of cis-W, tra -13 (which is formed from heating the transAQ, a.r-12isomer) and trans-%, cisAQ (from heating of the cis-9, trans-W isomer) may also be present.

3. Sialic Acid

Sialic acid is the term used to describe derivatives of neuraminic acid. Neuraminic acid is a nine-carbon length monosaccharide, found naturally in many animals, as well as plants, yeast, fungi and certain bacteria. In bacteria, sialic acids are commonly synthesised from mannose.

3.1 Sialyl Oligolac™ and Oligolac™

Sialyl Oligolac™ (SO) and Oligolac™ are derived from bovine colostrum. Sialyl Oligolac™ and Oligolac™ both contain enriched levels of sialic acids, lactose and numerous minerals, including calcium, potassium, sodium, phosphorus, magnesium and zinc.

Sialyl Oligolac™ and Oligolac™ are both prepared from bovine colostrum using the following method. Bovine colostrum milk is pasteurised and the lipid depleted. The substantially lipid depleted milk source is ultra filtered using a porous membrane and standard processing equipment. This provides a permeate absent large molecules such as casein, whey and

immunoglobulins. The retentate has a protein content of at least 50% expressed relative to the solids not fat (SNF) content of the retentate.

The permeate is subjected to a second filtration step using reverse osmosis. The retentate of the reverse osmosis is evaporated and dried to produce Oligolac™. Sialyl Oligolac™ is produced by subjecting the evaporated permeate of the Oligolac™ process to lactose reduction by crystallisation, and subsequent decanting, evaporation and drying.

4. Compositions useful in the present invention

A composition useful herein may be formulated as a food, functional food, drink, food additive, drink additive, dietary supplement, nutritional product, medical food, nutraceutical, medicament or pharmaceutical. Appropriate formulations may be prepared by an art-skilled worker with regard to that skill and the teaching of this specification.

In one embodiment the present invention relates to use of trans fatty acid or sialic acid in the manufacture of a food, functional food, drink, food additive, drink additive, dietary supplement, nutritional product, medical food, nutraceutical, medicament or pharmaceutical. In one

embodiment the trans fatty acid is CLA. In another embodiment the trans fatty acid is cis-9, transA 1 CLA. Preferably the composition is formulated for oral administration. Preferably the composition is for preventing or treating metabolic conditions, or other uses, as described above. In one embodiment the composition is in the form of a tablet, a caplet, a pill, a hard or soft capsule or a lozenge.

In one embodiment the composition is in the form of a cachet, a dispensable powder, granules, a suspension, an elixir, a liquid, a drink, or any other form that can be added to food or drink, including for example water or fruit juice. In one embodiment the composition is an enteral product, a solid enteral product or a liquid enteral product.

In one embodiment the composition further comprises one or more constituents (such as antioxidants) which prevent or reduce degradation of the composition during storage or after administration.

In one embodiment, compositions useful herein include any edible consumer product which is able to carry fats, fatty acids or lipid. Examples of suitable edible consumer products include baked goods, powders, liquids, confectionary products, reconstituted fruit products, snack bars, food bars muesli bars, spreads, sauces, dips, dairy products including ice creams, yoghurts and cheeses, drinks including dairy and non-dairy based drinks (such as milk drinks including milk shakes, and yogurt drinks), milk powders, sports supplements including dairy and non-dairy based sports supplements, food additives such as protein sprinkles and dietary supplement products including daily supplement tablets. Suitable nutraceutical compositions useful herein may be provided in similar forms. In preferred embodiments, the compositions useful herein are able to prevent or treat metabolic syndrome conditions in a subject to whom they are administered.

For example, such compositions are able to prevent or treat fatty liver disease in an obese, subject.

In another embodiment, compositions useful herein include dietetic products.

The term "dietetic product" means a product specially processed or formulated to satisfy particular dietary requirements which exist because of a particular physical or physiological condition and/ or specific diseases and disorders and which are presented as such.

For example, in subjects suffering from or susceptible to metabolic syndrome conditions, compositions of the present invention may be administered to modulate lipid or glucose levels in the subject. For example, to decrease serum or liver triglyceride levels.

In general, for oral administration a dietary (a food, food additive or food supplement for example), nutraceutical or pharmaceutical composition useful herein may be formulated by a skilled worker according to known formulation techniques.

Thus, a pharmaceutical composition useful according to the invention may be formulated with an appropriate pharmaceutically acceptable carrier (including excipients, diluents, auxiliaries, and combinations thereof) selected with regard to the intended route of administration and standard pharmaceutical practice. See for example, emington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed., Mack Publishing Co., 1980.

A dosage form useful herein may be administered orally as a powder, liquid, tablet or capsule. Suitable dosage forms may contain additional: agents as required, including emulsifying, antioxidant, flavouring or colouring agents, or have an enteric coating. Suitable enteric coatings are known. Enteric coatings surrounding the active ingredients and prevent the release of the active ingredients in the stomach but allow release after the dosage form has left the stomach. Dosage forms useful herein may be adapted for immediate, delayed, modified, sustained, pulsed or controlled release of the active components. Suitable formulations may contain additional agents as required, including emulsifying, antioxidant, flavouring or colouring agents.

Thus, the invention also is directed to doses, dosage forms, formulations, compositions comprising cis-9, trans-W CLA or sialic acid, including those disclosed herein, useful for the therapy of diseases, disorders, and/or conditions in humans and other mammals and other disorders as disclosed herein. The use of these dosage forms or formulations comprising one or more compositions of the invention enables effective treatment of these conditions. The dosage forms, ~ formulations, devices and/ or compositions of the invention may be formulated to optimize bioavailability, or to return or maintain plasma, blood, or tissue concentrations within the normal therapeutic range, including for extended periods. Controlled delivery preparations may also be used to optimize the antigen concentration at the site of action, for example.

The dosage forms, formulations or compositions of the invention may be formulated for periodic administration, for example to provide continued exposure to the one or more

compositions of the invention.

Examples of dosage forms suitable for oral administration include, but are not limited to tablets, capsules, lozenges, or like forms, or any liquid forms such as syrups, aqueous solutions, emulsions and the like, capable of providing a therapeutically effective amount of cis-9, trans-W CLA or sialic acid.

Examples of dosage forms suitable for buccal administration of the compositions and formulations of the invention include lozenges, tablets and the like, compositions comprising solutions and/ or suspensions in pharmaceutically acceptable, aqueous, or organic solvents, or mixtures thereof and/ or powders.

Further examples of dosage forms of the invention include, but are not limited to modified- release (MR) dosage forms including delayed-release (DR) forms; prolonged-action (PA) forms; controlled-release (CR) forms; extended-release (ER) forms; timed-release (TR) forms; and long- acting (LA) forms. For the most part, these terms are used to describe orally administered dosage forms, however these terms may be applicable to any of the dosage forms, formulations or compositions described herein. These formulations effect delayed total drug release for some time after drug administration, and/or drug release in small ahquots intermittently after administration, and/or drug release slowly at a controlled rate governed by the delivery system, and/or drug release at a constant rate that does not vary, and/ or drug release for a significantly longer period than usual formulations.

The compositions useful herein may be used alone or in combination with one or more other therapeutic agents. The therapeutic agent may be a food, drink, food additive, drink additive, food component, drink component, dietary supplement, nutritional product, medical food, nutraceutical, medicament or pharmaceutical.

In one embodiment the as-9, transA 1 CLA or sialic acid, optionally with at least one additional therapeutic agent are formulated for administration separately, simultaneously or sequentially.

In one embodiment the as-9, trans-W CLA or sialic acid, optionally with at least one additional therapeutic agent are formulated for co-administration.

In one embodiment the as-9, trans-W CLA or sialic acid, optionally with at least one additional therapeutic agent are formulated for sequential administration.

When used in combination with another therapeutic agent, the adixdnistration of a

composition useful herein and the other therapeutic agent may be simultaneous or sequential.

Simultaneous administration includes the administration of a single dosage form that comprises all components or the administration of separate dosage forms at substantially the same time.

Sequential administration includes administration according to different schedules, preferably so that there is an overlap in the periods during which the composition useful herein and other therapeutic agent are provided.

Additionally, it is contemplated that a composition in accordance with the invention may be formulated with additional active ingredients which may be of benefit to a subject in particular instances. For example, therapeutic agents that target the same or different facets of the disease process may be used.

It should be understood that the additional therapeutic agents may also be employed in a method according to the invention where they are administered separately, simultaneously or sequentially with a composition useful herein.

The efficacy of a composition useful according to the invention can be evaluated both in vitro and in vivo. Briefly, in one embodiment the composition can be tested for its ability, to for example, normalise or restore metabolic function in mtro. For in vivo studies, the composition can be fed to or administered to an animal (e.g., a mouse) and its effects on restoring metabolic function is then assessed. Based on the results, an appropriate dosage range and administration route can be determined.

The amount of as-9, transA 1 CLA or sialic acid to be administered is a therapeutically effective amount, which means an amount that is enough to elicit a desirable effect. A desirable effect is prevention or treatment of a condition associated with metabolic syndrome in a subject in need thereof.

As will be appreciated, the dose of the composition administered, the period of

administration, and the general administration regime may differ between subjects depending on such variables as the severity of symptoms of a subject, the type of disorder to be treated, the mode of administration chosen, the presence of additional components in the composition, such as minerals and the age, sex and/ or general health of a subject. However, by way of general example, the inventors contemplate administration of from about 15 mg to about 1200mg per kg body weight of a composition useful herein is administered per day, preferably about 15 to about 800 mg per kg per day. In one example, for a composition comprising sialic acid, a dosage rate of 15 to about 800 mg per kg per day is preferred. In another example, a composition comprising CLA is administered at a preferred dosage rate of 15 to about 800 mg per kg per day.

It should be appreciated that administration may include a single daily dose or administration of a number of discrete divided doses as may be appropriate. It should be understood that a person of ordinary skill in the art will be able without undue experimentation, having regard to that skill and this disclosure, to determine an effective dosage regime (including daily dose and timing of administration) for a given condition.

The present invention also relates to a dietary, nutraceutical or oral pharmaceutical

composition comprising, consisting essentially of or consisting of trans fatty acid, optionally in combination with a therapeutically active agent. In one embodiment the trans fatty acid is CLA. In another embodiment the trans fatty acid is as-9, trans-\ \ CLA.

In another aspect the present invention relates to a dietary, nutraceutical or oral

pharmaceutical composition comprising, consisting essentially of or consisting of sialic acid, optionally in combination with a therapeutically active agent.

In one embodiment a composition of the invention is a as-9, transA 1 CLA enriched milk fat. In one embodiment the ds-9, trans-W CLA enriched milk fat comprises at least about 1 , 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% by as-9, trans-W CLA, and useful ranges may be selected from any of these values (for example, from about 1 to about 99% by weight, from about 5 to about 99% by weight, from about 10 to about 99% by weight, from about 15 to about 99% by weight, from about 20 to about 99% by weight, from about 25 to about 99% by weight, from about 30 to about 99% by weight, from about 35 to about 99% by weight, from about 40 to about 99% by weight, from about 45 to about 99% by weight, from about 50 to about 99% by weight, from about 55 to about 99% by weight, from about 60 to about 99% by weight, from about 65 to about 99% by weight, from about 70 to about 99% by weight, from about 75 to about 99% by weight, from about 80 to about 99% by weight, from about 85 to about 99% by weight, from ^"about 90 to about 99% by weight, or from about 95 to about 99% by weight).

In one embodiment the composition comprises, consists essentially of or consists of about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, .80, 85, 90, 95 or 99% of ds-9, trans- 11 CLA and useful ranges may be selected between any of these foregoing values (for example, from about 1 to about 99%, from about 5 to about 99%, from about 10 to about 99%, from about 15 to about 99%, from about 20 to about 99%, from about 25 to about 99%, from about 30 to about

99%, from about 35 to about 99%, from about 40 to about 99%, from about 45 to about 99%, from about 50 to about 99%, from about 55 to about 99%, from about 60 to about 99%, from about 65 to about 99%, from about 70 to about 99%, from about 75 to about 99%, from about 80 to about 99%, from about 85 to about 99%, from about 90 to about 99%, or from about 95 to about 99% ).

In one embodiment the daily dosage range (by any route) is about 15 mg to about 1200mg per kg body weight of trans fatty acid; preferably about 15 to about 800 mg per kg per day. In another embodiment, the daily dosage range is about 15 mg to about 1200 mg per kg body weight of sialic acid, preferably 15 mg to about 800 mg per kg per day. A higher dose is preferred for short-term treatment and prevention and a lower dose for long-term treatment and prevention.

It should be understood that a person of ordinary skill in the art will be able without undue experimentation, having regard to that skill and this disclosure, to determine an effective amount of a composition of this invention for a given condition. ·. _ . .. _ 5. Metabolic syndrome and associated conditions

Metabolic syndrome is described as a cluster of metabolic disorders that increase the risk of developing cardiovascular disease and Type-2 diabetes mellitus. This cluster of disorders is characterised by central adiposity, adverse lipid and glucose profiles and hypertension. Metabolic syndrome is characterised by 3 or more symptoms from central obesity, elevated triglycerides, lowered HDL-cholesterol, elevated glucose, and elevated blood pressure (hypertension). Metabolic syndrome may lead to or is also associated with cardiovascular disease, atherosclerosis, non alcoholic fatty liver disease, type 2 diabetes mellitus, hyperinsulinaemia, insulin resistance, rnicroalbuminurea and hyperuricemia.

Markers used in the identification and characterisation of metabolic syndrome include increased triglyceride levels, increased postprandial lipemia, decreased high density lipoprotein (HDL) cholesterol, increased fasting glucose, and hypertension. In addition there may be associations with pro- flammatory and coagulation markers such as increased C-reactive protein, increased plasminogen activator inhibitor (PAI-1), and increased fibrinogen; also increased uric acid levels, and markers of poor liver function such as AST, ALT, GGT and ALP.

5.1 Non-alcoholie fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is a term used to describe a wide spectrum of liver damage, including hepatic steatosis, steatohepatitis, liver fibrosis and cirrhosis.

Non-alcoholic fatty liver disease is typically characterised by an accumulation of triglycerides in the liver, as a result of accelerated fatty acid mobilisation from visceral fat stofes that are deposited in the liver. In addition to this there is a decreased level of hepatic fatty acid β-oxidation and secretion of liver proteins from the liver. NAFLD encompasses not just lipid accumulation in the liver but also accompanying inflammation, fibrosis and tissue damage.

Various aspects of the invention will now be illustrated in non-limiting ways by reference to the following examples.

5.2 Markets useful in the identification of metabolic syndrome conditions

5.2.1 Body Mass Index

BMI is an index of adiposity based on an individual's height and weight. It is calculated as kg/ht². A BMI greater than 25kg/m² is indicative of overweight, while a measurement greater than 30kg/ m² is obese.

5.2.2 Central adiposity

Central adiposity, as measured via waist circumference, is characterised by the distribution of fat around the abdomen. Abdominal fat is reported to be highly correlated to an increased risk in cardiovascular disease.

5.2.3 Serum Uric Acid

Uric acid is produced by xanthine oxidase from xanthine and hypbxanthine. Elevated levels of uric acid are indicative of hyperuricemia, which is associated with a number of medical conditions including Lesch-Nyhan syndrome, insulin resistance, diabetes and cardiovascular disease.

5.2.4 Serum Triglyceride

Increased levels of triglycerides may be attributed to age, certain medications and medical conditions, or be heredity. Elevated triglycerides are indicative of a number of metabolic conditions, including metabolic syndrome, kidney and liver disease, hypothyroidism, diabetes and hypertension.

5.2.5 HDL cholesterol _. ,

High density lipoprotein cholesterol is reported to be involved in the prevention of atherosclerosis by removing cholesterol from, artery walls and transporting to the liver where it is metabolised. A low level of HDL cholesterol is indicative of increased risk of atherosclerosis and cardiovascular disease. 5.2.6 Post-prandial lipemia

Post-prandial lipemia is a term used to describe excess levels of lipids in blood following a fatty meal. Elevated blood lipid levels are indicative of coronary artery disease, thickening-of the carotid intimal, hypertension and cardiovascular disease.

5.2.7 Lipolytic activity

Lipolysis describes the breakdown of lipids by hydrolysis of triglycerides into its constituent free fatty acids. Consequently, decreased lipolytic activity results in a build up of triglycerides.

Diseases associated with decreased lipolytic activity include hypertriglyceridemia and diabetes.

5.2.8 C-reactive protein

C-reactive protein is a biomarker of inflammation and elevated levels in the blood are reported to be associated with proinflammatory states such as type 2 diabetes and cardiovascular disease.

5.2.9 Plasminogen activator inhibitor 1

Plasminogen activator inhibitor 1 inhibits the activity of plasminogen activator and urokinase, two enzymes involved in fibrinolysis. Inhibition of fibrinolysis results in decreased degradation of blood clots. An increased level of PAI-1 is indicative of coronary artery disease.

5.2.10 Fibrinogen

Fibrinogen or fibrin is a non-globular protein involved in blood coagulation. Increased fibrinogen production is associated with cardiovascular disease, whilst a decrease is associated with bleeding disorders.

5.2.11 Insulin resistance

Insulin resistance describes the condition wherein insulin becomes less effective at reducing blood glucose levels. Insulin resistance in fat and muscle results'in a -reduction in the uptake of glucose, while insulin resistance in liver cells results in reduced glycogen synthesis and storage. This causes a failure to suppress glucose production resulting in increased release into the blood. Insulin resistance leads to the development of Type-2 diabetes.

5.2.12 Hyperinsulinemia

Hyperinsulinemia is a condition characterised by an increased level of insulin in the blood. If left untreated, hyperinsulinemia can develop into diabetes.

5.2.13 Glucose

Increased level of fasting glucose is indicative of a number of metabolic diseases including insulin resistance and diabetes, and may be associated with cardiovascular disease. EXAMPLES

EXAMPLE 1 - In vitro indicators of efficacy of sialyl lactose and CLA-enriched milkfat against symptoms of metabolic syndrome.

This example describes the performance of in vitro experiments for two dairy derived compositions in laboratory models considered to be predictive of efficacy in preventing or treating metabolic syndrome.

Materials and Methods

A set of in vitro models well known to those skilled in the art as indicative of agent efficacy against aspects of metabolic syndrome were used to assess:

■ Uptake of Glucose by Skeletal Muscle, as described in Dieter et al. (2002) and Katz &

Westerblad (1995),

^■ Uptake of Glucose by Hepatocytes, as described in Seglen (1976) and Kreamer et al.

(1986).

^■ Uptake of Glucose by Adipocytes, as described in Xiangdong et al, (2001). ■ Production of Adiponectin by Epidydirnal Adipocytes, as described in Digirolamo, M. and Fine, J. B. (2001).

^■ Inflammation— Superoxide Production by Activated Neutrophils, as described in Tan, A.S. and Berridge, M.V. (2000). ^'

^■ Inflammation - TNF-a Production by Activated Neutrophils, as described in Greenspanm P. and Bauer, J.D. (2005).

^■ Inflammation - IL-1D Production from Monocyte/ Macrophage, as described in

Riedy, M.C. and Stewart, C.C. (1995).

^■ Nitric Oxide (NO) Production from Monocyte/Macrophage, as described in Riedy, M.C. and Stewart, C.C. (1995).

A set of ten dairy lipid fractions and ten dairy non-lipid fractions was selected to present to the in vitro testing models for characterisation for their potential to effect treatment of the symptoms of metabolic syndrome. Included in the set of lipid fractions was CLA-enhanced milkfat, a high trans-fatty acid milk fat fraction achieved by feeding milking cows a diet rich in linoleic acid.

Included in the set of non-lipid fractions was Sialyl Oligolac™, an enhanced sialic acid carbohydrate extract of bovine colostrum.

Each selected fraction was subjected to the set of in mtro screens to assess their impact on -these indicators of efficacy against the afflictions considered to be symptoms of metabolic syndrome.

Results The results for the fractions CLA-enriched milk fat (CLA-Mf) and Sialyl Oligolac™ (S.O.) are summarised in Table 1.

Table 1: The effect of CLA-enriched milk fat and Sialyl Oligolac™ on in vitro indicators of metabolic syndrome.

Neutr Neutr Neutr

Gluc- Glue Glue Insulin Macro Macro

Fraction SO - SO - SO - Hepat Adip(%) Soleu Control NO %400 TNFa

%100 - %200 %400

CLA-

- 57 108^† - 80^† 72^† 71^† 80++^† - Mf

S.O. 96 112 283+^† 143 87^† ' 98^† , 91^{† ■} > .62+ 189%

^* Positive result

The bovine high carbohydrate (CHO) fraction Sialyl Oligolac™ (SO) and the high lipid fraction conjugated linoleic acid (CLA) were shown to have glucose modulating and antiinflammatory effects and were selected as fractions for further investigation in an obese metabolic syndrome mouse model.

EXAMPLE 2 - The effect of dairy compositions on the health of mice fed a high fat diet

This example describes the efficacy of various dairy derived compositions in preventing metabolic syndrome in mice fed a high fat diet.

Materials and Methods

All animal experiments were approved by the appropriate ethics committee.

1. Mice and Diets

Male C57BL/ 6J weaner mice aged approximately 3 weeks were distributed into five groups (n=10 per group) as detailed in Table 4.

Two base diets were used, a low fat control AIN93G diet (Table 2) and a high fat control TD96132 diet (Table 3). Diets were supplemented for the trial groups as described below.

Table 2: Composition of low fat diet AIN93G

Ingredients Amount (g)

Corn Starch 7062.57

Casein 2994.54 ^'

Maltodextrin 2346..23

Sucrose 1777.26

Soybean Oil 314.43

Cellulose 748.64

AIN 93G Mineral Mix-(TD 94046) 524.05

AIN 93 Vitamin Mix (TD 94047) 149.73

L-Cystine 44.92

Choline Bitartrate 37.43 t-Butylhydroquinone (TBHQ) 0.21

TOTAL (g) 16000.00

Table 3: Composition of high fat diet TD96132

Ingredients Amount (g)

Corn Starch 2784.00

Casein 4106.40

Maltodextrin 2088.00

Sucrose 3177.59

Vegetable shortening, hydrogenated 1740.00

Beef Tallow 1740.00

Cellulose 696.00

AIN 93 G Mineral Mix (TD 94046) 718.62

AIN 93 Vitamin Mix (CA40060) 205.32

DL-Methionine 61.60

Calcium Phosphate, dibasic 82.13

Ethoxyquin (antioxidant) 0.35

TOTAL (g) 17400.00 . 2. Feeding Regime

There were three successive periods in the feeding trial, a 2 week lead-in, in which all animals were brought to a standardised state; a 15 week pre-treatment period, in which animals were conditioned to the required state for the treatment test, and an 8 week treatment period.

Group A (low fat control group) were fed the low fat control diet for the entire 25 week duration of the trial.

Group B (high fat control group) were fed the low-fat diet for 2 weeks, followed by the high- fat diet for the remaining 23 weeks.

Group C (sialyl lactose treatment group) were fed the low-fat diet for 2 weeks, followed by the high- fat diet until the end of week 17, and the high- fat diet supplemented with Sialyl Oligolac™ for the remaining 8 weeks.

Group D (enhanced trans-fat milkfat test group) were fed the low-fat diet for 2 weeks, followed by . the high- fat diet until the end of week 17, and the high-fat diet supplemented with CLA enriched milk fat for the remaining 8 weeks.

Group E (combined treatment test group) were fed the low-fat diet for 2 weeks, followed by the high-fat diet until the end of week 17, and the high-fat diet supplemented with Sialyl Oligolac™ plus CLA enriched milk fat for the remaining 8 weeks.

The diets were fed to . the mice ad libitum with normal drinking water.

Table 4: Diets fed to each group during the test period

Group A Control Low Fat - AIN 93G

Group B Control High Fat - TD96132

Group C TD96132 + sialyl oHgolac™ (3.5% w/w, 35g/kg)

TD96132 + CLA enriched milk fat

Group D

TD96132 + sialyl oligolac™ and CLA enriched milk

Group E

3. Body weight and food consumption measurements

Mice were monitored daily with body weight and food consumption measurements taken twice weekly throughout both trials.

Comparisons were made between the experimental (fraction-fed) groups and the control groups. 4. Blood collection

On days 15, 50, 85, 120, 148 and 176 of the trial, blood was collected from each experimental group mouse by tail tipping. Mice were fasted for 4 hours prior to blood collection.

Each serum sample was split into two aliquots (1 x 65uL for biochemical analysis and 1 x 35uL for BioPlexanalysis). Glucose, triglycerides, total cholesterol, HDL, and LDL levels were measured from the first aliquot. Insulin, TNFa, adiponectin and IL6 were measured from the second aliquot. All serum samples were prepared from each blood sample and frozen at -80°C.

In addition, blood was collected from all mice by cardiac puncture at the end of week 25. All mice were anaesthetised with ketamine (lOOmg/kg) and xylazine (lOmg/kg), punctured and the collected blood placed into heparin containing separating tubes. Following centrifugation, the plasma was aliquoted and stored at -20°C.

5. Faeces collection

Three days prior to each blood collection, a 24 hour collection of faeces took place.

Collections were weighed, and stored at -20°C for subsequent analysis of protein, carbohydrate and lipids.

6. Tissue analysis

Following blood collection by cardiac puncture at the completion of the trial, all mice were euthanised by C0₂ inhalation and cervical dislocation. All animals were autopsied and the skeletal muscle (quadriceps), liver (medial and lateral right lobes, medial and lateral left lobes and caudate lobe) were fixed in 4% formalin. Peri-renal fat, inguinal fat and epidydimal fat was removed and weighed.

7. Histological staining and image analysis

All tissues were snap frozen in liquid nitrogen and stored at -80°C, except for the Medial and Lateral Right Lobes and the -Caudate Lobe of each liver, which were fixed in 4% buffered formalin. The fixed tissues were placed in water to remove the formalin and transferred into 30% sucrose in PBS. The tissues were then frozen in optimal cutting temperature (OCT) compound (Tissue-Tek®, USA) and 5μιη sections cut using a Leica Cryostat (Model CM 3050S, Leica Microsystems GmbH, Germany). The sections were mounted onto microscope slides and allowed to dry before rinsing with 60% isopropanol and stained with 0.12% freshly prepared Oil Red O solution for 12 minutes. After staining, the slides were rinsed with 60% isopropanol, washed with distilled water and cover- slipped. Each slide was examined microscopically at a magnification of 100X and images captured using a Nikon DXM 1200 camera and Nikon ACT-1 software. Red pixel counts were obtained from each image. 8. Triglyceride assay

One piece of frozen liver tissue (approximately lOOmg) was minced and 2mL of

chloroform/methanol (2:1 v/v) added. The sample was homogenised for 2 minutes using a Polytron homogenizer (Polytron, Switzerland) and the sample transferred to a fresh tube and agitated for 2 hours on an orbital shaker. The homogenate was centrifuged at 500 x g for 0 minutes, and ImL of the liquid phase collected. The collected solvent was washed with 0.2mL of 0.9% NaCl solution, vortexed and centrifuged for a further 10 minutes at 225 x g. The upper phase containing the lipids was harvested and stored at -80°C for subsequent assaying. Measurements of triglyceride content was performed using a triglyceride kit (BioAssay Systems, USA) according the manufacturer's instructions. The concentration of triglyceride in the chloroform phase was converted to the content of triglyceride in liver tissue (mmol/g tissue).

9. Statistical analysis

Means and standard errors of the mean were calculated for each parameter and ANOVA used to compare each parameter. The Dunnet /-test was used to evaluate statistical significance with a ^' confidence level of p≤ 0.05.

Results

A decrease in the intensity of oil-red staining and a reduction in the size and number of lipid droplets evident in liver sections was observed in mice fed a high fat diet supplemented with Sialyl Oligolac™ or with CLA-enriched milk fat.

The individual response observed in mice fed a high fat diet supplemented with a combined therapy comprising both Sialyl Oligolac™ and CLA-enriched milk fat appeared quite varied. Four of the mice showed very low fat deposition, while two mice showed no fat deposition.

Test Groups C, D and E showed a significant reduction in the size and number of lipid droplets in liver sections compared to liver sections obtained from mice in Group B demonstrating a comparative beneficial effect for the dairy fractions in treating hepatic steatosis in the test animals.

This outcome justifies a confirmatory clinical trial in human subjects.

EXAMPLE 3— The effect of dairy compositions on the markers of metabolic syndrome and non-alcoholic fatty liver in overweight and obese individuals

This example describes the efficacy of dairy derived compositions in preventing metabolic syndrome in overweight and obese human subjects.. It is intended to show that Sialyl Oligolac™ and/or is-9 trans-W CLA administration improve markers of obesity-induced metabolic

dysregulation, including adverse liver function. . This study will be a 3-treatment randomised, crossover trial in a group of 30 overweight and obese male participants. Materials and Methods

Ethics approval will be obtained from the Northern X Regional Ethics Committee, Auckland, NZ. All participants will provide written consent and may withdraw from the trial at any time.

Inclusion/ exclusion criteria

The following criteria will be used to include or exclude individuals from participating in the trial: . ■

1.1 Inclusion criteria

Male gender

Aged 18-70 years

■ BMI > 25kg/m²

Central obesity (waist circumference >94cm, Asian ethnicity >90cm)

Abnormal liver function tests as indicated by raised liver enzyme(s) (i.e. AST >45 IU/L; ALT >45 IU/L; GGT >60 IU/L and ALP >120 IU/L) or raised serum ferritin

(>250mg/dL, with normal range Fe saturation)

■ Fatty liver as noted in liver ultrasound scan ^'

^■ Any one or more of the following adverse metabolic syndrome markers: Triglycerides

≥1.7mmol/L; HDL-cholesterol <1.03mmol/L; LDL-cholesterol≥3.4mmol/L; glucose ≥5.4mmol/L or blood pressure >130/85mmHg

^■ Change in body weight of < 10kg within the past 6 months

■ Willingness to participate

1.2 Exclusion criteria

^■ Allergy to milk or dairy products

^■ Endocrine, gastrointestinal (including liver), metabolic disease, cardiovascular events or cancer (including skin cancer), including prior history

■ Known hepatitis or other liver disease

^■ Significant renal impairment as indicated by serum creatinine >120μηιο1/Ε; eGFR

<60mL/min/ 1.73m²; and raised microalbuminurea (urine ACR >2.5mg/mol)

^■ Undergoing current treatment for metabolic disease, including hypertension, hyperlipidemia, Type-2 Diabetes Mellitus and gout

■ Currently undergoing a commercial weight loss regime or taking medications for weight loss

^■ Alcohol intake exceeding 30g/ day or >210g/week (equivalent to >3 standard drinks/ day or >21 standard drinks/week)

^■ Unwilling/unable to comply with trial protocol

^■ Participation in another clinical intervention trial 1.3 Recruitment

Thirty overweight male adult participants will be recruited from the general population within the Auckland city area through newspaper, poster and electronic advertisement. Individuals of all ethnicities will be welcome to participate in the metabolic study. Participants will provide properly executed written informed consent, in compliance with International Conference on Harmonization (ICH) guidelines.

2. Screening blood and urine analyses

During screening, participants will undergo full lipid profile, liver function tests, plasma glucose, serum creatinine and serum ferritin screens, full blood count, erythrocyte sedimentation rate and urine microalbuminurea screens.

3. Treatment regime

All participants will be randomly placed into three groups - Group A, Group B and Group C. Each group will complete the three trial arms, two treatments and a control, detailed in Table 5. Groups will be sequentially allocated to either active treatment or control for a period of 3 weeks, followed by a rmnimum 3 weeks wash out period between active treatments or control.

Table 5: Treatment regimes

Treatment 1 Sialy OHgolac (lOg/day, 138kJ) .

Treatment 2 CLA-enhanced Milkfat (4g/day, 138kJ)

Control Whole Milk Powder (6.5g/day. 138kJ)

A chocolate flavoured dairy dessert will be used as the

for administration of each of the dairy fractions. Participants will each receive approximately 150g daily dessert supplemented with the appropriate dairy treatment or control agent daily.

4. Administration of dairy fractions

Thirty participants (as selected according to the inclusion criteria detailed above) will complete a 12 hour fast prior to commencement of the trial and be randomly assigned to one of three treatments.

Following assignment to their initial treatment, they will daily consume over a three week period a 150g chocolate dairy dessert containing the dairy treatment Samples shall be single blinded as to their treatment so that subjects are unaware of what, if any, treatment they are consuming.

At the conclusion of the first three week treatment, subjects shall experience a three-week "washout" period in which no treatment is provided.

At the conclusion of the wash-out period each subject shall be assigned to a second of the three available treatments for the next three weeks, in the same manner as the first treatment was performed. Again, at the conclusion of the three-week second treatment period all subjects shall experience a three-week "wash-out" period in which no treatment is given. At the conclusion of the second wash-out period each subject shall be assigned to the remaining of three treatments that they have not already completed, and this final treatment period shall also last for three weeks. At the . conclusion of this third treatment subjects shall be released from the trial.

5. Treatment Assessments

During each three-week treatment period, participants will attend the clinic on 4 occasions (days 0, 7, 14 and 21). At each visit compliance will be assessed; body weight, waist and hip circumference, blood pressure measured; and a fasting blood sample collected. Adverse events and concomitant medications will also be recorded.

Blood samples will be analysed for typical indicators of metabolic syndrome, including fasting lipids (TAG, total cholesterol, LDL-C, HDL-C), liver function tests [AST, ALT, GGT, ALP], fasting glucose, fasting insulin, inflammatory markers (including hsCRP and ferritin) and serum uric acid.

Outcome variables will be analysed using Linear Mixed Model ANOVA (SPSS version 16.0,

SPSS Inc, Chicago, II, USA, 2007). No participants will be replaced due to drop out after randomization to treatment. Missing data will be assumed to be missing at random and no data imputation will be performed. Statistical significance will be set at a level of 0.05. Participant characteristics will be presented as mean, standard deviation (mean, SD). Efficacy endpoints will be presented as mean, standard error of the mean (mean, SEM).

Results

Data will show improved metabolic syndrome status for all, or at least some outcome variables for both the dairy treatments, Sialyl Oligolac™ and CLA-enriched milkfat when compared with the whole milk powder control.

Improved metabolic syndrome status is indicated by a change towards normal population norms for the short-term outcome variables, ie fasting lipids (TAG, total cholesterol, LDL-C, LIDL- C), liver function tests [AST, ALT, GGT, ALP], fasting glucose, fasting insulin, inflammatory markers (including hsCRP and ferritin) and serum uric acid. It is possible, but less likely that long- term indicators, body weight, waist and hip circumference, and blood pressure may also show improvements towards population norms during the trial period of the treatments.

INDUSTRIAL APPLICATION

The present invention has utility in treating or preventing conditions associated with metabolic syndrome. The described compositions may be employed as food or drink additives, nutritional products, dietary supplements, nutraceuticals and pharmaceuticals. The described compositions and methods of the invention may be employed to treat or prevent one or more of the conditions discussed herein.

Those persons skilled in the art will understand that the above description is provided by way of illustration only and that the invention is not limited thereto.

REFERENCES

Berry, M.N., Edwards, A.M. and Barritt, G.J., "Isolated hepatocytes preparation, properties and applications" In: Burdoh R.H. and Knippengerg, P.H.V. (Eds). Isolated hepatocytes preparation, properties and applications. New York: Elsevier Science Publishing, (1991) ppl 5-32

Dieter, N., Madar, Z. and Tirosh, O. "oc-Lipoic Acid Inhibits Glycogen Synthesis in Rat Soleus Muscle via Its Oxidative Activity and the Uncoupling of Mitochondria" /. Nutr. (2002) 132:3001- 3006.

Digirolamo, M. and Fine, J.B. "Cellularity measurements" In: Methods in Molecular Biology. Vol 155 : Adipose Tissue Protocols. Ailhaud G, Ed. Totowa, NJ, Humana Press (2001) p.65—75.

Greenspan, P., Bauer, J.D. et al. "Anti-inflammatory properties of the muscadine grape (Vitis rotundifolia): ] Agricultural <& Food Chemistry (2005) 53: 8481 - 8484

Katz, A. and Westerblad, H. "Insulin-mediated activation of glycogen synthase in isolated skeletal muscle: role of mitochondrial respiration". Biochim. Biophys. Acta (1995) 1244:229-232

Kreamer et al, Jn vitro cell and developmental Biology (1986) 22:201 -211

Riedy, M.C. and Stewart, C.C Current Protocol in Immunology, Supplement 13 CPI, Chapter 14: Macrophages and Monocytes. 14.5.1-14.5.6. John Wiley & Sons, Inc. (1995).

Seglen, Methods in ell Biology (1976) 13, 29-83

Tan, A.S. and Berridge, M.V. "Superoxide produced by activated neutrophils efficiently reduces the tetrazolium salt, WST-1 to produce a soluble formazan: a simple colorimetric assay for measuring respiratory burst activation and for screening of anti-inflammatory agents" / Immunol. Meth. (2000) 238: 59-68.

Tanti J-F, Cormont M, Gremeaux T, Le Marchand-Brustel Y. "Assays of glucose entry, glucose transporter amount, and translocation" In: Methods in Molecular Biology .Vol. 155 :

Adipose Tissue Protocols. Ailhaud G, Ed. Totowa, NJ, Humana Press (2001) p.157 -165

Wu, X., Motoshima, H., Mahadev, K., Stalker, T.J., Scalia, R. and Goldstein, B.J.

"Involvement of AMP-Activated Protein Kinase in Glucose Uptake Stimulated by the Globular Domain of Adiponectin in Primary Rat Adipocytes" Diabetes (2003) 52:1355-1363

Claims

. A method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of a trans fatty acid, or a salt, ester or precursor thereof.

2. The method of claim 1, wherein the trans fatty acid is present in a composition enriched in trans fatty acid. ^:

3. The method of claim 1, wherein the trans fatty acid is conjugated linoleic acid (CLA), or a salt, ester, or precursor thereof.

4. The method of claim 3, wherein the conjugated linoleic acid is selected from one or more CLA isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof.

5. The method of claim 4, wherein the CLA isomer is selected from the group comprising the cis-9,cis-l l isomer, the cis-9, trans-11 isomer, trans-9,cis-l 1 isomer, the trans-9,trans-l 1 isomer, the cis-10, cis-12 isomer, the cis-10, trans-12 isomer, the trans-10,cis-12 isomer, and the trans- 10, trans-12 isomer.

6. The method of claim 5, wherein the conjugated linoleic acid is cis-9, trans-\ \ CLA.

7. The method of any one of claims 1 to 6, wherein the trans fatty acid is a precursor to CLA, such as vaccenic acid.

8. The method of any one of claims 1 to 7, wherein the method comprises administration of cis- 9, transA 1 CLA and vaccenic acid.

9. The method of any one of claims 1 to 8, wherein the trans fatty acid is a trans 16:1 fatty acid, a -trans 18:1 fatty acid, or a combination thereof..

10. A method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of a trans fatty acid, or a salt, ester or precursor thereof.

11. The method of claim 10 wherein the trans fatty acid is CLA or a salt, ester, or precursor

thereof.

12. The method of claim 10 wherein the conjugated linoleic acid is selected from one or more CLA isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof.

13. The method of claim 12, wherein the CLA isomer is selected from the group comprising the cis-9,cis-l l isomer, the cis-9,trans-l 1 isomer, trans-9,cis-l 1 isomer, the trans-9,trans-l l isomer, the cis-10,cis-12 isomer, the cis-10,trans-12 isomer, the trans-10,cis-12 isomer, and the trans-10,trans-12 isomer.

14. The method of claim 11 wherein the conjugated linoleic acid is cis-9, trans-11 CLA.

15. The method of any one of claims 10 to 14 wherein the trans fatty acid is a precursor to CLA, such as vaccenic acid.

16. The method of any one of claims 10 to 15 wherein the method comprises administration of s-9, transA 1 CLA and vaccenic acid.

17. A method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of milk fat enriched with trans fatty acid.

18. The method of claim 17 wherein the trans fatty acid is CLA or a salt, ester, or- precursor

thereof.

19. The method of claim 18 wherein the conjugated linoleic acid is selected from one or more CLA isomers of 9,11-octadecadienoic acid and 10,12-octadecadienoic acid, in free or esterified form, or salts thereof, or mixtures thereof.

20. The method of claim 19 wherein the CLA isomer is selected from the group comprising the cis-9,cis-l l isomer, the cis-9,trans-l 1 isomer, trans-9,cis-l l isomer, the trans-9,trans-l l isomer, the cis-10,cis-l 2 isomer, the cis-10, trans- 12 isomer, the trans- 10,cis- 12 isomer, and the trans-10,trans-12 isomer.

21. The method of claim 18 wherein the conjugated linoleic acid is cis-9, trans-\ \ CLA.

22. The method of any one of claims 17 to 21 wherein the trans fatty acid is a precursor to CLA, such as vaccenic acid.

23. The method of any one of claims 17 to 22 wherein the milk fat or a composition in which the milk fat is present is enriched with both cis-9, trans-\ \ CLA and vaccenic acid.

24. The method of any one of claims 1 to 23 wherein the trans fatty acid, including the trans fatty acid present in the milk fat or in the composition comprises, consists essentially of, or consists of vaccenic acid.

25. The method of any one of claims 1 to 23 wherein the trans fatty acid, including the trans fatty acid present in the milk fat or in the composition comprises, consists essentially of, or consists of cis-9, transA 1 CLA.

26. The method of any one of claims 1 to 25 wherein the milk fat or a composition in which the milk fat is present is substantially free of the transAQ, cisA 2 CLA isomer.

27. The method of any one of claims 1 to 26 wherein the trans fatty acid, or the CLA-enriched milk fat comprises at least about 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50% by weight of cis-9, transA 1 CLA or a salt, ester or precursor thereof.

28. The method of claim 27 wherein the trans fatty acid or the milk fat includes trans-fatty acids comprising about 2 to 10% cis-9, transA 1 CLA or a salt, ester, or precursor thereof by weight.

29. The method of any one of claims 1 to 27 wherein the trans fatty acid or the milk fat comprises trans fatty acids which comprises at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% as- 9, transA 1 CLA or a salt, ester or precursor thereof by weight.

30. The method of claim 29 wherein the trans fatty acid or the milk fat includes trans fatty acids comprising about 70 to 80% as-9, trans-W CLA or a salt, ester or precursor thereof by weight.

31. The method of any one of claims 1 to 29 wherein the trans fatty acid or the milk fat comprises CLA isomers which comprise at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% CLA isomers by weight or a salt, ester or precursor thereof.

32. The method of claim 31 wherein the trans fatty acid or the milk fat includes CLA isomers comprising about 70 to 80% CLA isomers by weight.

33. The method of any one of claims 3 to 32 wherein the as-9, transA 1 CLA is selected from as-9, trans-W CLA derived from a natural source; synthetic as-9, trans-W CLA; as-9, trans- CLA in free fatty acid form; as-9, transA 1 CLA bound to glycerol, a monoglyceride, a diglyceride or a triglyceride; as-9, transA 1 CLA in esterified form; or mixtures thereof.

34. The method of any one of claims 7 to 33 wherein the milk fat is produced by enhancing natural levels of CLA in milk by feeding a milk producing mammal with a diet enriched in at least one fatty acid (e.g. linoleic acid).

35. The method of any one of claims 17 to 33 wherein the milk fat or the composition is prepared by combining a source of as-9, transA 1 CLA or a salt, ester or precursor thereof with milk fat.

36. A method of preventing or treating a metabolic syndrome condition, the method comprising administering to a subject in need thereof an effective amount of a composition comprising sialic acid.

37. A method of preventing or treating fatty liver disease, the method comprising administering to a subject in need thereof an effective amount of a composition comprising sialic acid.

38. The method of claim 36 or 37 wherein the composition comprises bound sialic acid and free sialic acid.

39. The method of any one of claims 36 to 38. wherein the composition comprises sialic acid and at least one monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharide.

40. The method of any one of claims 36 to 39 wherein at least some of the sialic acid is bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharides.

41. The method of any one of claims 36 to 40 wherein at least some of the sialic acid is covalentiy bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharides.

42. The method of any one of claims 36 to 41 wherein the disaccharide is lactose.

43. The method of any one of claims 36 to 42 wherein the sialic acid is covalentiy bound to the lactose.

44. The method of any one of claims 36 to 43 wherein the composition comprises sialic acid and lactose, wherein the sialic acid is administered with lactose either simultaneously, separately or sequentially.

45. The method of any one of claims 36 to 44 wherein the composition additionally comprises minerals.

46. The method of claim 45 wherein the minerals are selected from the group comprising calcium, potassium, sodium, phosphorus, magnesium or zinc.

47. The method of any one of claims 36 to 46 wherein the composition additionally comprises phospholipids.

48. - The method of claim 47 wherein the phospholipid is selected from the group comprising phosphatidylethanolamine, phosphatidylmositol or sphingomyelin.

49. The method of any one of claims 2 to 48 wherein the composition comprises at least about 0.2. 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5% sialic acid.

50. The method of claim 49 wherein the composition comprises about 1% to 2% sialic acid.

51. The method of any one of claims 2 to 50 wherein the composition comprises at least about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 or 90% lactose.

52. The method of claim 51 wherein the composition comprises about 55% to 65% lactose.

53. The method of any one of claims 2 to 52 wherein the composition comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, 0, 11 or 12% protein. ,

54. The method of claim 53 wherein the composition comprises about 6% to 10% protein.

55. The method of any one of claims 2 to 54 wherein the composition comprises 2, 4, 6, 7, 8, 10, 12, 14, 16, 18 or 20% minerals.

56. The method of claim 55 wherein the composition comprises about 10% to 18% minerals.

57. The method of any one of claims 2 to 56 wherein the composition comprises from about 40% to about 90% lactose and from about 0.2% to about 5% sialic acid.

58. The method of claim 57 wherein the composition comprises from about 50% to about 70% lactose and from about 1% to about 3% sialic acid.

59. The method of claim 57 wherein the composition comprises from about 70% to about 90% lactose and from about 0.2% to about 0.5% sialic acid.

60. The method of claim 57 wherein the composition comprises from about 40% to about 80% lactose, from about 1% to about 5% sialic acid and from about 4% to about 18% minerals.

61. The method of claim 57 wherein the composition comprises from about 55% to about 65% lactose, from about 1% to about 2% sialic acid and from about 5% to about 18% minerals.

62. The use of a trans fatty acid or a salt, ester or precursor thereof, in the preparation-of a medicament for use in preventing or treating a metabolic syndrome condition in a subject.

63. The use of sialic acid or a composition comprising sialic acid in the preparation of a

medicament for use in preventing or treating a metabolic syndrome condition in a subject.

64. The use of claim 62 wherein the medicament additionally comprises sialic acid.

65. The use of claim 63 or 64 wherein the medicament comprises bound sialic acid and free sialic acid.

66. The use of any one of claims 62 to 65 wherein the medicament comprises sialic acid and at least one monosaccharide, disaccharide, trisacchaiide, polysaccharide or oligosaccharide.

67. The use of any one of claims 63 to 66 wherein at least some of the sialic acid is bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisacchande, polysaccharide or oligosaccharides. . . .

68. The use of any one of claims 63 to 67 wherein at least some of the sialic acid is covalently bound to at least a portion of the one or more of the monosaccharide, disaccharide, trisaccharide, polysaccharide or oligosaccharides.

69. The use of claim 68 wherein the disaccharide is lactose.

70. The use of claim 69 wherein the sialic acid is covalently bound to the lactose.

71. The use of claim 69 wherein the medicament comprises sialic acid and lactose, wherein the sialic acid is administered with lactose either simultaneously, separately or sequentially.

72. The use of any one of claims 62 to 71 wherein the medicament additionally comprises

minerals.

73. The use of claim 72 wherein the minerals are selected from the group comprising calcium, potassium, sodium, phosphorus, magnesium or zinc.

74. The use of claim 73 wherein the composition additionally comprises phospholipids.

75. The use of claim 74 wherein the phospholipid is selected from the group comprising

phosphatidylethanolamine, phosphatidylinositol or sphingomyelin.

76. The use of any one of claims 62 to 75 wherein the medicament comprises at least about 0.2, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5% sialic acid.

77. The use of claim 62 wherein the medicament comprises about 1% to 2% sialic acid.

78. The use of any one of claims 62 to 77 wherein the medicament comprises at least about 40, ■ 45, 50, 55, 60, 65, 70, 75, 80, 85 or 90% lactose.

79. The use of claim 78 wherein the medicament comprises about 55% to 65% lactose.

80. The use of any one of claims 62 to 79 wherein the medicament comprises at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12% protein.

81. The use of claim 80 wherein the medicament comprises about 6% to 10% protein.

82. The use of any one of claims 62 to 81 wherein the medicament comprises 2, 4, 6, 7, 8, 10, 12, 14, 16, 18 or 20% minerals.

83. The use of claim 82 wherein the medicament comprises about 10% to 18% minerals.

84. The use of any one of claims 62 to 83 wherein the medicament comprises from about 40% to about 90% lactose and from about 0.2% to about 5% sialic acid.

85. The use of claim 84 wherein the medicament comprises from about 50% to about 70%

lactose and from about 1 % to about 3% sialic acid.

86. The use of claim 84 wherein the medicament comprises from about 70% to about 90%

lactose and from about 0.2% to about 0.5% sialic acid.

87. - The use of claim 84 wherein the medicament comprises from about 40% to about 80%

lactose, from about 0.5% to about 5% sialic acid and from about 4% to about 18% minerals.

88. The use of claim 84 wherein the medicament comprises from about 55% to about 65%

lactose, from about 0.5% to about 2% sialic acid and from about 5% to about 18% minerals.

89. A composition comprising a trans fatty acid and one or more additional agents for use in preventing or treating a metabolic syndrome condition in a subject in need thereof.

90. The composition of claim 89 wherein the trans fatty acid is CLA or a salt, ester, or a precursor to CLA, such as vaccenic acid.

91. The composition of claim 89 or 90 wherein the trans fatty acid is ds-9, transA X CLA and

vaccenic acid.

92. The composition any one of claims 89 to 91 wherein the one or more additional agents are selected from the group comprising lactose, sialic acid, one or more minerals, and one or more proteins.

93. A composition comprising sialic acid and optionally one or more additional agents for use in preventing or treating a metabolic syndrome condition in a subject in need thereof.

94. The composition of claim 93 wherein the additional agent is a trans fatty acid, ds-9, trans A 1 CLA or vaccenic acid.

95. The composition of any one of claims 93 to 94 wherein the composition comprises sialic acid and trans fatty acid.

96. The composition of any one of claims 89 to 95 wherein an additional agent is milk fat

enriched with at least one trans fatty acid, including ds-9, transA 1 CLA or vaccenic acid.

97. The method of any one of claims 1 to 61 , the use of any one of claims 62 to 88, or the

composition of any one of claims 89 to 96 wherein the metabolic syndrome condition is a condition selected from the group comprising central adiposity, obesity, dyslipidemia, cardiovascular disease, atherosclerosis, hypertension, Type-2 diabetes mellitus, hyperinsulinemia, insulin resistance, hyperglycaemia, non-alcoholic fatty liver disease, nonalcoholic steatohepatitis, microalbuminuria or hyperuricemia.

98. The method of any one of claims 1 to 61, the use of any one of claims 62 to 88, or the

composition of any one of claims 89 to 96 wherein the metabolic syndrome condition is characterised by one or more of the following: increased uric acid levels, increased triglyceride levels, increased postprandial lipemia, decreased high density lipoprotein cholesterol,

^" decreased post-heparin lypolytic activity, increased sympathetic nervous system activity, increased sodium retention, increased C-reactive protein (CRP), increased plasminogen activator inhibitor (PAI-1), increased fibrinogen, or increased fasting glucose.

99. The method, use or composition of any one of the preceding claims wherein the effective amount, the medicament or the composition is formulated as a food, drink, food additive, drink additive, dietary supplement, nutritional product, powder, liquid, food bar, spread, sauce, ointment, tablet, capsule, medicament, pharmaceutical or nutraceutical.

100. The method, use or composition of any one of the preceding claims wherein the effective amount, the medicament or the composition is formulated for enteral administration, such as oral administration.

101. The method, use or composition of any one of the preceding claims wherein the effective amount, the medicament or the composition is formulated as a pharmaceutical composition suitable for enteral administration, such as oral administration.