WO2024026529A1 - Preparation for the treatment of non-alcoholic fatty liver disease (nafld) - Google Patents

Abstract

Provided herein is a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of non-alcoholic fatty liver disease (NAFLD). Another embodiment of the invention includes a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain. One further embodiment of the invention includes the use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for prophylaxis and/or treatment of the effects of NAFLD.

Description

PREPARATION FOR THE TREATMENT OF NONALCOHOLIC FATTY LIVER DISEASE (NAFLD)

Related Application

[001] The present application claims convention priority from Australian provisional patent application 2022902151, filed on 1 August 2022. The content of AU’ 151 is incorporated herein by reference in its entirety.

Field of the Invention

[002] The invention relates to the field of food supplement manufacture and therapeutic manufacture. In particular, the invention relates to a combination of virgin sugarcane prebiotic and probiotic strains, the synbiotic combination of the combination in the diet of an individual, and the improved outcomes of the synbiotic approach in the prophylaxis and/or treatment and management of non-alcoholic fatty liver disease (NAFLD) risk factors.

[003] Although the present invention will be described hereinafter with reference to its preferred embodiment, it will be appreciated by those of skill in the art that the spirit and scope of the invention may be embodied in many other forms.

Background of the Invention

[004] Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

[005] Non-alcoholic fatty liver disease (NAFLD) is characterised by excessive fat accumulation, i.e., steatosis, without significant alcohol consumption. Liver steatosis is defined as fat accumulation in >5% of hepatocytes [1]. NAFLD can be sub-divided into two types - simple steatosis or non-alcoholic fatty liver (NAFL) without inflammation and non-alcoholic steatohepatitis (NASH). In individuals who consume little or no alcohol, NAFL can progress from simple liver steatosis (fatty liver) to NASH. The latter (due to inflammation in the liver and liver damage) has a potential progressive course leading to liver fibrosis, cirrhosis and hepatocellular carcinoma [2].

[006] NAFLD is the most common cause of chronic liver diseases in Western countries, affecting up to 40% of general population and sometimes reaching 90% [3]. The clinical burden of NAFLD is not only confined to liver-specific morbidity and mortality, but there is growing evidence that NAFLD is a multi-system disease affecting extra-hepatic organs and regulatory pathways. NAFLD increases risk of type-2 diabetes mellitus (T2DM), cardiovascular disease and chronic kidney diseases [4].

[007] The global prevalence of NAFLD in general population is estimated to be 25% [2]. NAFLD is closely associated with obesity-related metabolic disorders such as insulin resistance, T2DM, hypertension and hyperlipidaemia, and is even recognised as the hepatic component of metabolic syndrome [2] [3] [4]. NAFLD and metabolic syndrome both increase the risk of cardiovascular disease and T2DM and are therefore likely to have similar risk profiles [3]. Hence, unsurprisingly, the leading cause of death in NAFLD patients is not liver failure, but cardiovascular disease [5]. The prevalence of NAFLD is 80-90% in obese adults, 30-50% in patients with diabetes and up to 90% in patients with hyperlipidaemia. The prevalence of NAFLD among children is 3-10%, rising up to 40-70% among obese children [6] .

[008] NAFLD pathogenesis is complex and not fully understood. It is, however, understood that NAFLD is caused by complex interplay of multiple factors including poor diet, insulin resistance, and other metabolic disturbances. Additional risk factors include host factors (genetics and epigenetics) and lifestyle -related factors like sleep shortage, irregular food intake, sedentary habits, and excessive weight gain [7].

[009] There are currently no FDA- approved drugs to treat NAFLD [8] and conventional antibiotics are prescribed to reduce the symptoms. However, these can cause side effects and potential bacterial resistance. Available drugs such as vitamin E, pioglitazone, and pentoxifylline have borderline efficacy, but are limited by potential side-effects and toxicities, and do not improve liver fibrosis. Weight gain is common in patients taking thiazolidinediones, and these drugs can cause fluid retention and precipitate congestive heart failure [9]. The primary therapeutic options include weight loss and the combination of healthy diet and exercise. Therefore, novel interventions are sought that can target the underlying risk factors particularly gut microbial dysbiosis and the inflammation associated therewith [7].

[0010] The gut microbiome includes several trillion bacteria with a combined bacterial genome of 100 times as many genes as the human genome. Key roles assigned to gut microbiota include maintenance of mucosal integrity and defence, development of innate immunity, and energy harvest from dietary sources. Alterations in the gut microbiome have been linked to a range of illnesses, including obesity and NAFLD.

[0011] NAFLD is commonly associated with compositional and functional alterations of gut microbiota known as dysbiosis. Gut microbiota analysis in NAFLD compared with healthy subjects noted reduced commensal bacteria abundance in the gram-positive Firmicute phylum and increased opportunistic pathobionts such as those from the gramnegative Proteobacterial phylum [3] [5].

[0012] The gut dysbiosis in NASH promotes a proinflammatory environment, which results in increased gut permeability. Increased gut permeability can cause the translocation of endotoxin lipopolysaccharides (LPS), bacteria, and antigens into the portal circulation and thence into the liver, resulting in hepatic inflammation [10]. Inflammation, which is the key pathological characteristic of NASH, is a major mediator promoting the development of NASH and progression to cirrhosis and hepatocellular carcinoma. Many factors could be involved in the NASH inflammatory process, including genetics, epigenetic, environmental agents, nutritional, and adverse shifts in the gut microbiota [10]. [0013] The association with adverse shifts in the gut microbiota [3][11] has prompted research into alternative treatments aimed at modulation of gut microbiota [12] [13]. In this context, probiotic and prebiotic components that can influence gut microbiota, strengthen intestinal barrier integrity, and modulate immune response, provide strategies to attenuate inflammation. The baseline treatment and avoidance of inflammatory events can be approached from a dietary perspective and studies demonstrate a role for prebiotics, probiotics and the synergistic combination administration, labelled synbiotics as microbial therapies.

[0014] Synbiotics, being a combination of probiotic and prebiotic ingredients that interact positively, can potentiate prophylactic and therapeutic effects that may function synergistically to confer health benefits. This is critical when selecting probiotic and prebiotic ingredients for synbiotic combination for efficient functioning and significant efficacy.

[0015] Many prebiotics are purified fibres of limited chemical complexity and do not present the multiple range of glycosidic bond types that occur naturally in fruit and vegetables. Developing prebiotics that replicate the complexity of nature’s chemical structures has not been standard practise in commercially-available products to date. However, providing a simple prebiotic that is readily fermented by gut microbiome may not be the correct strategy given it could result in a natural selection towards bacteria that thrive on the simplified fibre structures and unbalance the diversity that is known to be an important factor in optimal function of the bowel and protection from disease.

Additionally, rapid fermentation of prebiotic fibre is known to accompany gastrointestinal symptoms including abdominal pain, bloating and gas.

[0016] A unique prebiotic phytonutrient fibre extract from sugarcane, manufactured to preserve the cell wall components provides a representative cellular fibre component from plant dietary sources. While sugarcane fibre has historically been labelled as an insoluble fibre, improved fibre analysis techniques and studies into the fermentability of sugarcane fibrous matter reveals uniform fermentability of the insoluble fibre characteristics and fibrous sub-structures by bacteria, specifically those that reside and can function in the colon. Providing this complex and more nature-equivalent nutrient source to the microorganisms of the colon, including functional delivery of sugarcane’s characterised antioxidant bioactives (phenolics, flavonoids, and polycosanols), delivers a positive pressure on maintaining or increasing microbial diversity, rather than the reduction of diversity with limited complexity prebiotic s.

[0017] Similarly, several factors influence the beneficial effects of probiotics. Their survival in delivery formats, including functional foods, is required. They must also survive during gastric transit to elicit health effects of the live organism. An additional factor that may influence the beneficial effects of probiotics is their ability to generate fermentation products that influence the composition and function of the gut microbiome. This, in turn, can potentially improve the health of the host. Bacteriocins and organic acids are two possible antimicrobial products that can be produced by probiotics to influence and stabilise the gut microbiome. Other intermediate metabolites, including short-chain fatty acids (SCFAs) produced as a consequence of bacterial fermentation of prebiotics in the gut, have been affirmed to exert beneficial effects on the host.

[0018] The probiotic effect could, therefore, be potentiated by co- supplementation with prebiotic dietary fibres that can be metabolised and fermented by the administered probiotic, as well as by beneficial microflora in the gut. The resulting production of metabolites can direct the shift of immune markers from pro-inflammatory to antiinflammatory phenotype. Accordingly, a synbiotic approach potentially offers greater success of colonisation and survivability of beneficial bacteria owing to the advantageous effects that can be achieved compared to using either probiotics or prebiotics alone [14][15], [0019] By targeting the normalisation of gut microbiota, the production of beneficial byproducts of microbial fermentation (such as short chain fatty acids), the delivery of plant antioxidants, and supplying a known beneficial probiotic, the synergistic effects can be optimised beyond the sum of its parts.

[0020] Kfibre® is a complex prebiotic dietary fibre sourced from virgin sugarcane with the sugar removed and has been reported to influence gut health in several studies involving inflammatory bowel disease (IBD) mouse models, human GERD/acid reflux trials and animal/human in vitro faecal studies.

[0021] Research on mice models of gut inflammation found that preconditioning the gut with Kfibre significantly lowered the severity of IBD parameters and reduced inflammatory immune responses to a chemical irritant (prophylactic effect) and reduced an established gut inflammation (therapeutic effect) in a genetic model of mucin abnormality [14][15],

[0022] Kfibre in a synergistic combination with probiotic (as synbiotic supplement) is also confirmed to amplify the anti-inflammatory and immunomodulatory effects in both models of IBD. In both gut inflammatory models of IBD, Kfibre-based synbiotic administration was confirmed to prevent and/or enhance the gut barrier integrity, reduce pro -inflammatory markers in the serum (CRP and cytokines), increase SCFA production along the colon and modulate microbiota diversity. In another nutritionist led Kfibre weight loss study, Kfibre supplementation has been reported to cause significant weight loss (10.06 kg in females and 9.97 kg in males) compared to those who were not administered Kfibre (5.70 and 6.41 kg, respectively).

[0023] The sugarcane fibre has been previously demonstrated to impart positive effects on human gut microbiota in vitro, particularly influencing the abundance of members of family Bifidobacteriaceae [16]. Compared with the other dietary fibre supplements tested (wheat dextrin and psyllium husk), the sugarcane fibre was reported to show highest availability of polyphenols and antioxidant potential.

[0024] In a double-blind, randomised, placebo-controlled pilot clinical study, 3-week consumption of virgin sugarcane prebiotic fibre relative to placebo (cellulose) showed marked reduction in gastroesophageal reflux diseases (GERD) symptoms (heartbum, regurgitation, and total symptoms scores) in medically-diagnosed GERD patients [17]. [0025] Kfibre is sourced from freshly cut sugarcane and utilises only water and mechanical stress in the processing to the final product (chemical free and low heat stress). It is marketed as a complex, raw un-purified prebiotic dietary fibre for gut health management. Anecdotal consumer feedback and unpublished trial data indicated that it could have a range of benefits countering bloating, indigestion, constipation, and food intolerances as well as improving blood sugar control. A human trial evaluating a range of NAFLD- related clinical parameters is needed to be able to understand the impact that Kfibre & Kfibre-based synbiotic on NAFLD. This approach would provide publishable evidence to support the positive role of Kfibre in combination with probiotic for alleviating NAFLD. [0026] It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

[0027] It is an object of an especially preferred form of the present invention to provide a synergistic combination of prebiotic sugarcane fibre and probiotics in the diet of an individual for the prophylaxis and/or treatment of NAFLD.

[0028] Another object of an another especially preferred form of the present invention is to provide a synergistic combination of a prebiotic fibre extract from sugarcane and probiotics that ameliorates the effects/inflammatory states of NAFLD, as a long-term prophylactic treatment by inclusion into the regular diet.

[0029] Although the invention will be described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms.

Definitions and nomenclature

[0030] Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

[0031] As used herein, a “synbiotic” is a combination of prebiotic plant fibre from sugarcane and a probiotic bacterial strain or postbiotic byproduct for use in all subsequent formats, formulations and purposes. The synbiotic in most cases, includes but is not limited to, a combination of dry prebiotic and dry probiotic spores, lyophilised or live stabilised bacteria. The synbiotic can also be the combination in an aqueous solution for use in food manufacturing procedures.

[0032] A “postbiotic” should be construed in accordance with the definition ascribed to it by the International Scientific Association for Probiotics and Probiotics (ISAPP), which in 2019 convened a panel of experts specialising in nutrition, microbial physiology, gastroenterology, paediatrics, food science and microbiology to provide a consistent and credible definition for the term. The panel defined a “postbiotic” as a “preparation of inanimate microorganisms and/or their components that confers a health benefit on the host”. Effective postbiotics must contain inactivated microbial cells or cell components, with or without metabolites, that contribute to observed health benefits. The panel also discussed existing evidence of health-promoting effects of postbiotics, potential mechanisms of action, levels of evidence required to meet the stated definition, safety and implications for stakeholders. The panel determined that a definition of postbiotics is useful so that scientists, clinical triallists, industry, regulators and consumers have common ground for future activity in this area, see, https://www.nature.com/articles/s41575-021- 00440-6.

[0033] A “carrier” is a palatable substrate for the sugarcane fibre, which may or may not contain protein or other nutrients. The carrier can be in a solid or liquid form, including but not limited to: fruit extracts, broths, purees, dairy products, baked goods.

[0034] An “inert filler’ is any product used to increase the bulk of fibre according to the invention to allow for ease of handling by the user. The filler may contain flavours or nutrients, and other dietary fibres to improve mouth feel, but does not necessarily contribute to the total benefit provided by the invention.

[0035] A “pellet” includes any compact form of the invention, including but not limited to: a dried pill or tablet in the manner of a vitamin or a “soft lolly” style lozenge that may be used as a treat or as an addition to other foods.

[0036] As used herein, the phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. When the phrase “consists of’ (or variations thereof) appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole. As used herein, the phrase “consisting essentially of’ limits the scope of a claim to the specified elements or method steps, plus those that do not materially affect the basis and novel characteristic(s) of the claimed subject matter.

[0037] With respect to the terms “comprising”, “consisting of’, and “consisting essentially of’, where one of these three terms is used herein, the presently disclosed and claimed subject matter may include the use of either of the other two terms. Thus, in some embodiments not otherwise explicitly recited, any instance of “comprising” may be replaced by “consisting of’ or, alternatively, by “consisting essentially of’.

[0038] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as modified in all instances by the term “about”, having regard to normal tolerances in the art. The examples are not intended to limit the scope of the invention. In what follows, or where otherwise indicated, “%” will mean “weight %”, “ratio” will mean “weight ratio” and “parts” will mean “weight parts”.

[0039] The term “substantially” as used herein shall mean comprising more than 50% by weight, where relevant, unless otherwise indicated.

[0040] The term “about” should be construed by the skilled addressee having regard to normal tolerances in the relevant art.

[0041] The recitation of a numerical range using endpoints includes all numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

[0042] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the invention. [0043] It must also be noted that, as used in the specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0044] As used herein, a “heaped teaspoon” of Kfibre® is 1.5 g. Therefore, when taken twice a day, the daily amount of Kfibre® is 3 g.

[0045] The probiotic (L. plantarum 299v (DSM 9843)) is generally taken once daily for the first five days, and then twice daily thereafter. Metagenics Ultra Flora GI Soothe is a commercial preparation comprising 20 billion CFU per capsule, resulting in a daily dosage (post day 5) of 40 billion CFU.

[0046] The ratio of prebiotic to probiotic is therefore 1.5 to 3 g prebiotic to 20-40 billion CFU probiotic for live probiotic bacterial cells.

[0047] Akkermansia muciniphila is a human intestinal symbiont, isolated from human feces. It is a mucin-degrading bacterium belonging to the genus, Akkermansia. It belongs to the phylum Verrucomicrobiota and its type strain is Muc^T (=ATCC BAA-835^T =CIP 107961^T). [0048] Any of the features described herein can be combined in any combination with any one or more of the other features described herein within the scope of the invention.

Summary of the Invention

[0049] The present invention relates generally to the combination of a prebiotic sugarcane fibre and probiotic strains, and the synbiotic use of the combination in the diet of an individual for the prophylaxis and/or treatment of NAFLD.

[0050] According to the first aspect of the present invention, there is provided a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

[0051] It is particularly preferred that the prebiotic phytonutrient fibre material prepared from virgin sugarcane is Kfibre®.

[0052] In an embodiment, the prebiotic phytonutrient fibre material prepared from virgin sugarcane, which is washed and chopped, and the sugary juice removed. The leftover plant is dried and ground into a fibre powder. Around 95% of the sugar is removed.

[0053] In an embodiment, the prebiotic phytonutrient fibre material prepared from virgin sugarcane, freshly harvested and manufactured to retain phytonutrients and intact cell wall vegetable fibre. The resultant product is 100% sugarcane fibre (95% sucrose removed) which has a composition of <87% fibre. The other 13% comprises natural sugars, waxes, minerals, vitamins, plants sterols and antioxidants.

[0054] Preferably, the prebiotic phytonutrient fibre material is prepared via a process including the steps of: subjecting sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the fibre, and to enhance the water retention properties of the residual fibre product. Preferably, the wet diffusion step is a diffusion extraction, performed under relatively low-shear conditions. Preferably, the wet diffusion step is performed within the temperature range of about 25 °C to about 70 °C.

[0055] The composition can further comprise a pharmaceutically acceptable carrier, solvent, base and/or excipient.

[0056] There are a number of advantages to using a prebiotic fibre material prepared from virgin sugarcane in the manner described above. Firstly, there are no known adverse allergic effects associated with this source. Secondly, as a fibre source it more accurately represents vegetables than purified or isolated fibre sources prepared from grains or chemically manufactured from other sources. Thirdly, it is high in essential micronutrients and phytonutrients and has the ability to protect micronutrients from other foods when consumed in conjunction with a meal.

[0057] Sugarcane fibre prepared using the steps described above also has several advantageous properties compared to purified/isolated plant fibres such as bran, psyllium husk and inulin. The fibre prepared from sugarcane is a true lignose, hemicellulose and cellulose combination, like the total dietary fibres found in most vegetables. Pectin is a significant component (about 10%) of cell walls, and being a soluble, metabolisable fibre, could also play a role in the activity of the sugarcane fibre as a prebiotic. Other components of the sugarcane fibre include xylan (which is soluble) and arabinoxylan polymers.

[0058] The sugarcane fibre can be classed as almost entirely insoluble fibre, using standard chemical methods of classification, however, it has many of the properties of soluble fibres such as high water-binding capacity (up to 8-10 times by weight) and a prebiotic effect by acting as a substrate for microbial fermentation in the colon to generate beneficial metabolites including SCFAs.

[0059] This combination of both soluble and insoluble fibre characteristics as well as the retention of phytonutrients provides for a food profile that closely mimics an ideal diet. [0060] The at least one probiotic bacterial strain can be commercially sourced from bacterial strains that are known and correlated to the reduction of chronic gut inflammation or to the support of the normal function of the gastrointestinal tract. Preferably, at least one probiotic bacterial strain is spore-forming, thereby providing robustness to survive and retaining viability in the colon of a host.

[0061] Alternatively, the at least one probiotic bacterial strain can be heat stable to ensure viability during manufacture.

[0062] In a further alternative, the at least one bacterial strain can be acid stable, allowing the at least one bacterial strain to remain viable in the host colon after administration.

[0063] In certain embodiments, the at least one probiotic bacterial strain is spore-forming, heat stable and acid stable.

[0064] Preferably, the at least one probiotic bacterial strain is selected from the group consisting of: Bacillus coagulans, Lactobacilli (L. plantarum, L. paracasei, L. acidophilus, L. casei, L.rhamnosus, L. crispatus, L. gasseri, L. reuteri, L. bulgaricus), Bifidobacterium (B. longum, B. catenulatum , B. breve, B. animalis, B. bifidum), Streptococcus (S. sanguis, S. oralis, S. mitis, S. thermophilus, S. salivarius), Bacillus (B. subtilis, B. laterosporus), Lactococcus (L. lactis), Enterococcus (E. faecium), Pediococcus (P. acidilactici), Propionibacterium (P. jensenii, P. freudenreichii), Pep to streptococcus (P. productus), Saccharomyces (S. boulardii) and Akkermansia muciniphila.

[0065] According to a second aspect of the present invention, there is provided use of a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

[0066] According to a third aspect of the present invention, there is provided a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

[0067] According to a fourth aspect of the present invention, there is provided use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for prophylaxis and/or treatment of the effects of NAFLD.

[0068] According to a fifth aspect of the present invention, there is provided use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for ameliorating the effects of NAFLD.

[0069] According to a sixth aspect of the present invention, there is provided a composition for ameliorating the effects of NAFLD; said composition comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

[0070] According to a seventh aspect of the present invention, there is provided use of a food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane.

[0071] According to an eighth aspect of the present invention, there is provided a food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

[0072] The food product can be utilised in the diet for a defined period of time for ameliorating the effects of NAFLD. Alternatively, the food product can be utilised in the diet over a prolonged period of time, including indefinitely, to ameliorate the effects of NAFLD and/or as a prophylactic for NAFLD.

[0073] According to a ninth aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0074] According to a tenth aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0075] According to an eleventh aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0076] According to a twelfth aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0077] According to a thirteenth aspect of the present invention, there is provided a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic. In the context of this invention, a “postbiotic” is a non-viable bacterial product or metabolic by-product from probiotic microorganisms that has biologic activity in the host. In other words, a postbiotic is a fragment or metabolite of a probiotic. [0078] The composition can further comprise a pharmaceutically acceptable carrier, solvent, base and/or excipient.

[0079] The at least one postbiotic can be commercially-sourced. Preferably, the at least one postbiotic is sufficiently robust to survive in the colon. Alternatively, the at least one postbiotic can be heat stable to ensure viability during manufacture. In a further alternative, the at least one postbiotic can be acid stable, allowing the at least one postbiotic to remain viable in the colon after administration. In certain embodiments, the at least one postbiotic is heat stable and acid stable.

[0080] The at least one postbiotic can be selected from the group consisting of short-chain fatty acids, antimicrobial peptides, nutrients (including amino acids and vitamins such as vitamin K and B-vitamins), carbohydrate-active enzymes.

[0081] According to a fourteenth aspect of the present invention, there is provided a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic.

[0082] According to a fifteenth aspect of the present invention, there is provided a composition for ameliorating the effects of NAFLD; said composition comprising prebiotic fibre material prepared from sugarcane, and at least one postbiotic.

[0083] According to a sixteenth aspect of the present invention, there is provided a food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one postbiotic.

[0084] The food product can be utilised in the diet for a defined period of time for ameliorating the effects of NAFLD. Alternatively, the food product can be utilised in the diet over a prolonged period of time, including indefinitely, to ameliorate the effects of NAFLD and/or as a prophylactic for NAFLD.

[0085] According to a seventeenth aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0086] According to an eighteenth aspect of the present invention, there is provided use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

[0087] According to a nineteenth aspect of the present invention, there is provided a method of treating or ameliorating the effects of NAFLD in a subject, the method comprising administering to the subject a composition provided by the second or thirteenth aspects, or a food product provided by the third or fourteenth aspects.

[0088] Any of the features described herein can be combined in any combination with any one or more of the other features described herein within the scope of the invention.

Brief Description of the Drawings

[0089] A preferred embodiment of the present invention will now be described with reference to the accompanying drawings, in which:

[0090] Figure 1 is a graph showing the effects of the synbiotic on serum GGT of NAFLD subjects 1-7 as described below. The red bar represents gamma-glutamyl transferase (GGT) before treatment with the synbiotic (t=0) and the green bar represents GGT after synbiotic treatment (t=6 weeks). In all cases, GGT decreased significantly, and generally in the region of 30-60%.

[0091] Figure 2 is a graph showing the effect of the synbiotic on body weight of NAFLD subjects 1-7 as described below. The red bar represents body weight before treatment with the synbiotic (t=0) and the green bar represents body weight after synbiotic treatment (t=6 weeks). In all cases, body weight decreased significantly, and generally in the region of 10-20%.

Detailed Description of the Invention

[0092] The present invention takes advantage of the properties of a prebiotic fibre produced from sugarcane in such a way that maximises retention and minimises destruction of the bioactive molecules. The present inventors have surprisingly found that such a prebiotic fibre is effective in the prophylaxis and/or treatment of inflammatory states associated with NAFLD.

[0093] It has also been surprisingly found that combining the prebiotic fibre with a probiotic or a postbiotic results in a synbiotic effect, whereby inflammatory states are reduced by more than when the prebiotic, probiotic or postbiotic is used in isolation, and more than just the additive effect of the prebiotic and probiotic, or the prebiotic and postbiotic.

[0094] The method of preparation of the prebiotic fibre material from sugarcane is broadly similar to that described in International Patent Publication WO 2011/035381, to KFSU Pty Ltd; this document is incorporated herein by reference in its entirety. However, the preparatory method according to the present invention can be generally described in the following steps: a sugarcane size reduction step; a relatively gentle aqueous extraction step that separates the fibre from other sugarcane fractions, including the sugar fraction, without causing degradation of the fibre functionality; and a relatively gentle drying step that minimises degradation of the fibre functionality.

[0095] It is preferred that the aqueous extraction step be an aqueous diffusion extraction performed at about neutral pH. It is also preferred that the drying step be a rapid vortex drying operation that can be achieved via a low temperature vortex dryer, the dryer being able to reduce the wet weight of the sugarcane material from 40-80% wet weight to less than 10% wet weight in 10-30 seconds while not heating the material to a level that would significantly damage the bioactives in the plant material.

[0096] The invention provides for the use of a synbiotic, namely, a prebiotic fibre from sugarcane combined with selected probiotics or postbiotics, in the formulation of foods, diets or therapeutics that reduces the risk of development of NAFLD conditions, or which ameliorates the symptoms of those conditions.

[0097] When prepared according to the invention, the synbiotic combination has a number of advantages over other fibre sources and food, including that: it is relatively hypoallergenic; it contains both insoluble and bound soluble fibre in beneficial proportions for dietary intake; it can be prepared in a ‘chemical-free’ manner and contains no harmful trace elements, unlike fibre from other sources such as chemically modified starch; it can be prepared in such a way as to retain the micronutrients and active molecules found in the “molasses” component of sugarcane, without the need to extract and purify those components for their biological function; and it can be prepared in such a manner to optimize the bioactivity of the prebiotic and also maximise viability of the probiotic or its by-products (postbiotics).

[0098] It is also known that too much fibre in the diet can have several negative side effects including but not limited to constipation, diarrhoea and bad flatulence.

[0099] Advantageously, in embodiments of the present invention, where the fibre product is added as a cellular-based fibre supplement to a subject’s diet, dietary fibre intake can be more easily controlled with a sterically hindered restricted rate of microbial metabolism.

Examples

[00100] The following working embodiments of the synbiotic of the invention are contemplated by the Applicant:

Example 1

[00101] In this example, 0.5-5.0 g of a synbiotic according to the invention is pressed into a pellet or added to a flavouring medium and pressed into a pellet. Due to the inherent stability of the prebiotic and selected probiotics, the synbiotic pellets are prepared at a formulation level such that the dose may be varied according to a patient’s requirements to reduce the symptoms of NAFLD without incurring negative effects.

Example 2

[00102] In this example, a synbiotic is mixed with a flavoured drink (for example a non-acidic fruit juice or milk) and pasteurised (1-5 g per 100-250 mL). A drink prepared in this manner is a convenient, ready-to-consume product for the amelioration of NAFLD. In an embodiment, pasteurisation sterilises the preparation.

Example 3

[00103] In this example, a supplement is prepared as an easy-to-measure powder with or without flavours, stabilisers and inert filler, formulated specifically to be combined with water. Specifically, a synbiotic can be mixed with a dry flavour component and an inert filler to form easy-to-use granules. The dose (e.g., 1-5 g) can be in a convenient single-serve sachet or in a multi-dose bulk pack. The resultant supplement is best suited to allow reduced meal size, as the granules can be mixed with water (thereby allowing less food to be consumed each meal).

Example 4

[00104] In this example, a synbiotic is prepared in a solid flavoured meal such as a biscuit, a bar, or a bread (baked) product (0.5-5.0 g per serve of ready mixed food). Multiple biscuits, bars or bread products can be consumed by an individual to provide a specific dosing regimen and optimise compliance of the treatment. This has three principal advantages over other delivery systems in that it feels more like a treat for the consumer, it eliminates the need for liquid, and reduces the total volume of the stomach contents.

Additionally, the increased saliva production may have a complementary effect with the synbiotic benefits.

Example 5

[00105] In this example, seven subjects with non-alcoholic fatty liver disease, were prescribed a combination of virgin sugarcane prebiotic (e.g., Kfibre® as described above) and a specific probiotic strain L. plantarum 299v (DSM 9843) as well as general lifestyle and dietary advice over six weeks. The subjects were assessed before and after six weeks of supplementation with the synbiotic combination for positive effects by way of beneficial weight management and serum levels of the NAFLD biomarker, gamma-glutamyl transferase (GGT). The subjects use and outcomes are reported below:

[00106] Subject 1 [AM] - 50-year-old female with medically diagnosed fatty liver and reflux, overweight and with prescribed medication of Coversyl (perindopril) 5 mg daily. In addition, the subject wished to lose weight and was seeking help with bloating.

[00107] The subject was prescribed one heaped teaspoon of Kfibre twice a day, 229v probiotic once daily for five days, then twice a day along with her medication of Coversyl 5 mg daily. The subject was recommended to drink 2 L filtered water daily and follow a wellness diet plan, exercise three times a week and have outside time in nature at least once a week. After six weeks, the GGT levels in the subject dropped from 73 to 50 U/L and she experienced significant weight loss of 78 into 68 kg.

[00108] Subject 2 [AM] - 49-year-old male with medically diagnosed fatty liver where ultrasound showed infiltration of liver by fat deposits and overweight.

[00109] The subject was prescribed one heaped teaspoon Kfibre twice a day, 229v probiotic one daily for five days then twice a day, one tablespoon collagen powder and one tablespoon whey powder once a day, male multivitamin once a day, plus a general lifestyle and dietary advice similar to that of Subject 1. The subject also followed the Metagenics keto diet eating plan. After six weeks, the blood GGT level of the subject dropped from 92 to 43 U/L and the subject experienced weight loss of 120 into 109 kg.

[00110] Subject 3 [VK] - 53-year-old female with medically diagnosed fatty liver, reflux, overweight, perimenopause and with prescribed medication of Avapro (irbesartan) 150 mg once daily.

[00111] In addition to her current medication of Avapro, the subject was prescribed one heaped teaspoon Kfibre twice a day, 229v probiotic once daily for five days then twice a day, plus ovainne (female multivitamin with vitex/chaste tree) in addition to a general lifestyle and dietary advice similar to that of Subject 1. After six weeks, the subject’s blood GGT levels dropped from 65 to 40 U/L and the subject experienced weight loss of 114 into 98 kg.

[00112] Subject 4[SV] - 32-year-old female with medically diagnosed fatty liver, overweight, with bloating and experience tiredness and with history of reflux.

[00113] The subject was prescribed one heaped teaspoon Kfibre twice a day, 229v probiotic once daily for five days then twice a day, plus female multivitamins and mineral, magtabs (magnesium 200 mg with zinc 10 mg) daily, in addition to following a general lifestyle and dietary advice similar to that of Subject 1. After six weeks, the subject’s blood GGT levels dropped from 87 to 53 U/L and the subject experienced weight loss of 69 into 60 kg.

[00114] Subject 5 [MF] - 55-year-old female with medically diagnosed fatty liver, reflux, cyst on kidney, overweight and experience tiredness. The subject also reported bloating with too much milk/dairy and was allergic to penicillin.

[00115] In addition to continuing medication of Avapro HCT (irbesartan and hydrochlorothiazide) 150 mg once daily, the subject was prescribed one heaped teaspoon Kfibre twice a day, 229v probiotic once daily for five days then twice a day. The subject was cautioned to avoid dairy. The subject was additionally prescribed prime essential (multimineral and vitamin with Vitamin E), CoQlO 150 mg each once daily, mega magnesium tablet once daily plus followed general lifestyle and dietary advice similar to that of Subject 1. After six weeks, the subject’s blood GGT levels dropped from 67 to 55 U/L and the subject experienced weight of 79 into 75 kg.

[00116] Subject 6[AG] - 38-y ear-old male with medically diagnosed fatty liver, GAD (generalised anxiety disorder), reflux, cyst on liver, overweight and experiencing tiredness, panic attacks, and was out of breath at times.

[00117] In addition to continuing medication of Effexor (venlafaxine) 75mg once daily, the subject was prescribed one heaped teaspoon Kfibre twice a day, 229v probiotic once daily for five days then twice a day. The subject was additionally prescribed Infla- mood (Metagenics; saffron and turmeric) one, twice a day, biopractica genomulti (activated B) and additionally followed general lifestyle and dietary advice similar to that of Subject 1. After six weeks, the subject’s blood GGT levels dropped from 96 to 42 U/L and the subject experienced weight loss of 83 into 75 kg.

[00118] Subject 7 [FS] - 65-year-old male with medically diagnosed fatty liver, reflux, panic attacks, osteoarthritis and experience tiredness, panic attacks and has bad taste in mouth.

[00119] In addition to continuing medications of Effexor (venlafaxine) 150 mg and Coversyl (perindopril) 5 mg, the subject was prescribed one heaped teaspoon of Kfibre twice a day, 229v probiotic once daily for five days then twice a day. The subject was additionally prescribed prostoco (saw palmetto, stinging nettle, hawthorn) once daily and followed general lifestyle and dietary advice similar to that of Subject 1. Subject was also reported to go swimming every day with a personal trainer, worked out three times a week and saw an occupational therapist. After six weeks, the subject’s blood GGT levels dropped from 95 to 39 U/L and the subject experiences significant weight loss of 115 into 106 kg.

[00120] The benefits propounded by the seven subjects are significant and demonstrate that supplements prepared according to the invention directly counter NAFLD as seen from the reduction of blood biomarker GGT and benefits to weight management. All subjects have indicated they are certain that the invention is the source of improvement in their respective pathologies.

[00121] The data obtained from a nutritionist-led NAFLD study using Kfibre in combination with specific probiotic bacteria, L. plantarum 299v. The study involving NAFLD patients where supplementation of Kfibre was in combination with L. plantarum 299v has been observed to significantly reduce the NAFLD parameters, particularly serum GGT and excessive body weight. Applicant has engaged with the nutritionist to acquire these clinical-quality data to review, interpret and extend outcomes in patent applications and in further randomised controlled trials in terms of best probiotic compatibility with Kfibre. These data will form the basis for running a targeted human clinical trial using Kfibre and specific probiotic in combination (synbiotic) to alleviate NAFLD risk factors and for management of NAFLD.

[00122] The gut microbiota has a significant impact on intrahepatic fat accumulation; however, the mechanisms are yet to be fully determined. The interaction between the liver and gut, that is, the gut-liver axis, could explain the beneficial role of gut microbiota composition for NAFLD. Applicant’s evidence suggests that targeting the axis using prebiotics, probiotics or synbiotics may be an appropriate approach for treating NAFLD and for management of its risk factors.

[00123] It will be understood by those skilled in the art that the above examples represent merely some of the ways that the invention may be put into effect. Other embodiments may be envisaged that, while differing in some details, nevertheless fall within the scope of the invention.

References

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Statements of Invention

[00124] The following statements of invention are contemplated by the Applicant in accordance with the disclosure made above:

1. A composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

2. A composition according to statement 1, wherein each probiotic bacterial strain is present as spores and/or live cells.

3. A composition according to statement 1, wherein the composition is in a form selected from the group consisting of a pellet, food product, and drink.

4. A composition according to statement 1, wherein the prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

5. A composition according to statement 1, further comprising a pharmaceutically acceptable carrier, solvent, base or excipient.

6. A composition according to statement 1, wherein said prebiotic phytonutrient fibre material comprises almost the entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

7. A composition according to statement 1, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

8. Use of a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

9. Use according to statement 8, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

10. Use according to statement 8, wherein each said probiotic bacterial strain is present as spores and/or live cells.

11. Use according to statement 8, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

12. Use according to statement 8, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

13. Use according to statement 8, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

14. A food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

15. A food product according to statement 14, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

16. A food product according to statement 14, wherein each said probiotic bacterial strain is present as spores and/or live cells.

17. A food product according to statement 14, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

18. A food product according to statement 14, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

19. A food product according to statement 14, wherein the prebio tic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

20. Use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for prophylaxis and/or treatment of the effects of NAFLD.

21. Use according to statement 20, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

22. Use according to statement 20, wherein each said probiotic bacterial strain is present as spores and/or live cells.

23. Use according to statement 20, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

24. Use according to statement 20, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

25. Use according to statement 20, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. Use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for ameliorating the effects of NAFLD. Use according to statement 26, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. Use according to statement 26, wherein each said probiotic bacterial strain is present as spores and/or live cells. Use according to statement 26, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. Use according to statement 26, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. Use according to statement 26, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. A composition for ameliorating the effects of NAFLD; the composition comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain. A composition according to statement 32, wherein each probiotic bacterial strain is present as spores and/or live cells. 34. A composition according to statement 32, wherein the composition is in a form selected from the group consisting of a pellet, food product, and drink.

35. A composition according to statement 32, wherein the prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

36. A composition according to statement 32, further comprising a pharmaceutically acceptable carrier, solvent, base or excipient.

37. A composition according to statement 32, wherein said prebiotic phytonutrient fibre material comprises almost the entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

38. A composition according to statement 32, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

39. Use of a food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane.

40. Use according to statement 39, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

41. Use according to statement 39, wherein each said probiotic bacterial strain is present as spores and/or live cells.

42. Use according to statement 39, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. 43. Use according to statement 39, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

44. Use according to statement 39, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

45. A food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

46. A food product according to statement 45, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

47. A food product according to statement 45, wherein each said probiotic bacterial strain is present as spores and/or live cells.

48. A food product according to statement 45, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

49. A food product according to statement 45, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

50. A food product according to statement 45, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. Use according to statement 51, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. Use according to statement 51, wherein each said probiotic bacterial strain is present as spores and/or live cells. Use according to statement 51, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. Use according to statement 51, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. Use according to statement 51, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. Use according to statement 57, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. Use according to statement 57, wherein each said probiotic bacterial strain is present as spores and/or live cells. Use according to statement 57, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. Use according to statement 57, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. Use according to statement 57, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. Use according to statement 63, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. Use according to statement 63, wherein each said probiotic bacterial strain is present as spores and/or live cells. 66. Use according to statement 63, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

67. Use according to statement 63, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

68. Use according to statement 63, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

69. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

70. Use according to statement 63, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

71. Use according to statement 63, wherein each said probiotic bacterial strain is present as spores and/or live cells.

72. Use according to statement 63, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

73. Use according to statement 63, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. Use according to statement 63, wherein the prebio tic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. A composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic. A composition according to statement 75, wherein each probiotic bacterial strain is present as spores and/or live cells. A composition according to statement 75, wherein the composition is in a form selected from the group consisting of a pellet, food product, and drink. A composition according to statement 75, wherein the prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. A composition according to statement 75, further comprising a pharmaceutically acceptable carrier, solvent, base or excipient. A composition according to statement 75, wherein said prebiotic phytonutrient fibre material comprises almost the entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. A composition according to statement 75, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. A food product comprising a prebiotic phytonutrient fibre material extracted from sugarcane, and at least one postbiotic. A food product according to statement 82, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. A food product according to statement 82, wherein each said probiotic bacterial strain is present as spores and/or live cells. A food product according to statement 82, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. A food product according to statement 82, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. A food product according to statement 82, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. A composition for ameliorating the effects of NAFLD; said composition comprising prebiotic fibre material extracted from sugarcane, and at least one postbiotic. A composition according to statement 88, wherein each probiotic bacterial strain is present as spores and/or live cells. 90. A composition according to statement 88, wherein the composition is in a form selected from the group consisting of a pellet, food product, and drink.

91. A composition according to statement 88, wherein the prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

92. A composition according to statement 88, further comprising a pharmaceutically acceptable carrier, solvent, base or excipient.

93. A composition according to statement 88, wherein said prebiotic phytonutrient fibre material comprises almost the entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

94. A composition according to statement 88, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

95. A food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one postbiotic.

96. A food product according to statement 95, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

97. A food product according to statement 95, wherein each said probiotic bacterial strain is present as spores and/or live cells.

98. A food product according to statement 95, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. 99. A food product according to statement 95, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

100. A food product according to statement 95, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material.

101. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

102. Use according to statement 101, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink.

103. Use according to statement 101, wherein each said probiotic bacterial strain is present as spores and/or live cells.

104. Use according to statement 101, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre.

105. Use according to statement 101, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers.

106. Use according to statement 101, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. Use according to statement 107, wherein the prebiotic phytonutrient fibre material is administrable in a form selected from the group consisting of a composition, pellet, food product, and drink. Use according to statement 107, wherein each said probiotic bacterial strain is present as spores and/or live cells. Use according to statement 107, wherein said prebiotic phytonutrient fibre material is sucrose-reduced sugarcane fibre. Use according to statement 107, wherein said prebiotic phytonutrient fibre material comprises almost an entire insoluble fibre content of sugarcane, comprising a lignose, hemicellulose and cellulose combination, including pectin, xylan and arabinoxylan polymers. Use according to statement 107, wherein the prebiotic phytonutrient fibre material is prepared using a process comprising the steps of: subjecting the sugarcane to at least one wet diffusion step to separate sugars from a residual fibre material whilst maintaining nutrient content; and, subjecting the residual fibre material to a rapid, low-heat drying process to retain biologically active molecules in the residual fibre material and to enhance water retention properties of said residual fibre material, to thereby produce the prebiotic phytonutrient fibre material. 113. A method of treating or ameliorating the effects of NAFLD in a subject, the method comprising administering to the subject a composition defined according to any one of statements 1-7, 32-38, 75-81 or 88-94, or a food product as defined according to any one of statements 14-19, 45-50, 82-87 or 95-100. 114. A composition, use, method or food product according to any one of the preceding statements, wherein the ratio of prebiotic to probiotic in the synbiotic is between about 1.5 to 3.0 g prebiotic to between about 20 to 40 billion CFU probiotic.

[00125] Although the invention has been described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms.

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS

1. A composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

2. Use of a food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain for prophylaxis and/or treatment of the effects of NAFLD.

3. A food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

4. Use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for prophylaxis and/or treatment of the effects of NAFLD.

5. Use of a composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, for ameliorating the effects of NAFLD.

6. A composition for ameliorating the effects of NAFLD; the composition comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

7. Use of a food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane.

8. A food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one probiotic bacterial strain.

9. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

10. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

11. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

12. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one probiotic bacterial strain, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect.

13. A composition comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic.

14. A food product comprising a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic.

15. A composition for ameliorating the effects of NAFLD; said composition comprising prebiotic fibre material prepared from sugarcane, and at least one postbiotic.

16. A food product for ameliorating the effects of NAFLD; said food product comprising prebiotic fibre material prepared from sugarcane, and at least one postbiotic. . Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a therapeutic for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. Use of a prebiotic phytonutrient fibre material prepared from sugarcane, and at least one postbiotic, in the manufacture of a food product for ameliorating the symptoms and/or inflammatory states of NAFLD, or for providing a prophylactic effect. A method of treating or ameliorating the effects of NAFLD in a subject, the method comprising administering to the subject a composition defined according to any one of claims 1, 6, 13 or 15, or a food product as defined according to any one of claims 3, 8, 14 or 16. A composition, use, method or food product according to any one of the preceding claims, wherein the ratio of prebiotic to probiotic in the synbiotic is between about 1.5 to 3.0 g prebiotic to between about 20 to 40 billion CFU probiotic.