Microencapsulates and uses thereof

Abstract

A slow-release microencapsulate comprises a FODMAP encapsulated within a denatured protein matrix. The microencapsulate is gastric-resistant and ileal-sensitive, and configured to release the FODMAP slowly in the ileum of the subject to inhibit or prevent gas formation and bloating. A method of preventing or inhibiting gastrointestinal gas formation or bloating associated with consumption of FODMAP in a subject is also described.

Description

[0001] TITLE

[0002] Microencapsulates and uses thereof

[0003] Technical Field

[0004] The present invention relates to microencapsulates, and uses thereof.

[0005] Technical Background

[0006] A prebiotic is defined as a nondigestible compound that, through its metabolization by microorganisms in the gut, modulates the composition and / or activity of the gut microbiota, thus conferring a beneficial physiologic effect on the host. Health benefits ascribed to prebiotic dietary in the literature include increases in Bifidobacteria and Lactobacilli, production of beneficial metabolites .increases in calcium absorption, decreases in protein fermentation, decreases in pathogenic bacteria populations, decreases in allergy risk, positive effects on gut barrier permeability, and improved immune system defense. Examples of prebiotics include dietary fibres including fructans (including inulin and fructooligosaccharide (FOS)), gluten and galactooligosaccharides (GOS), non-digestible monosaccharides and disaccharides, and non-digestible polyols. The group of nondigestible saccharides are known as FODMAP’s (fermentable oligosaccharides, disaccharides, monosaccharides and polyols).

[0007] While FODMAP’s are beneficial and provide prebiotic benefits, many of the oligosaccharides are relatively short chains with an average degree of polymerization between 3-10. Due to this short chain length they are very quickly fermented by the bacteria in the gut that causes a dose-dependent significant gas formation in a relatively short time period that cause gas formation and bloating, and other diarrhoea or constipation in patients who are sensitive. They are also known to exacerbate the symptoms of irritable bowel syndrome (IBS) in people with a highly sensitive gut.

[0008] It is an object of the disclosure to overcome at least one of the above-referenced problems.

[0009] Summary of the Disclosure

[0010] The applicant has addressed the objective by providing prebiotic fibre and other FODMAP in a microencapsulated form in which the microencapsulates are configured to pass through the stomach intact and release their contents slowly in the ileum. This has the benefit of releasing the FODMAP slowly in the ileum, reducing sudden gas formation and bloating. In addition, the applicant has discovered that microencapsulation of prebiotic dietary fibre has the added benefit of stabilising prebiotic dietary fibre in low pH, heat processed products (such as a carbonated beverages containing prebiotic dietary fibre) by inhibiting or preventing hydrolysis of the dietary fibre to simple sugars. Method of microencapsulation of probiotic bacteria and other nutrients are described in the literature and include emulsion, cold gelation and fluidised bed microencapsulation methods (as described in more detail below).

[0011] In a first aspect, there is provided a microencapsulate comprising a FODMAP encapsulated within a polymer (e.g. protein) matrix, in which the microencapsulate is gastric-resistant and ileal-sensitive.

[0012] In any embodiment, the polymer is denatured protein.

[0013] In any embodiment, the denatured protein comprises denatured plant or dairy protein.

[0014] In any embodiment, the FODMAP is a prebiotic dietary fibre is selected from a fructan (e.g. a |3[2, 1 ]-fructan), a beta-glucan, a galactooligosaccharides (GOS), an isomaltooligosaccharide, lactulose, xylooligosaccharide and arabinooligosaccharide.

[0015] In any embodiment, the fructan is selected from a fructooligosaccharide (FOS), oligofructose and inulin.

[0016] In any embodiment, the protein matrix is configured to release the FODMAP in the ileum over a period of 1 to 6 hours.

[0017] In any embodiment, the microencapsulate has a dimension of 50 to 500 pm.

[0018] In any embodiment, the microencapsulate consists essentially of one or more FODMAP’s (e.g. a prebiotic dietary fibre) and a protein matrix.

[0019] In any embodiment, the microencapsulate has a core-shell morphology.

[0020] In any embodiment, the shell comprises the polymer matrix.

[0021] In any embodiment, the core comprises (or consists essentially of) prebiotic dietary fibre.

[0022] In any embodiment, the microencapsulate does not contain any bacteria.

[0023] In a second aspect, there is provided a composition comprising a multiplicity of microencapsulates according to the disclosure.

[0024] In any embodiment, the composition is selected from an edible product such as a food, beverage, or nutritional supplement. Examples include dairy products, carbonated beverages, confectionary products, and cereal products. In any embodiment, the composition comprises a low FODMAP food product and a multiplicity of microencapsulates according to the disclosure. The term “Low FODMAP foods” refers to food products that contain minimal amounts of fermentable carbohydrates that can trigger digestive symptoms in people with irritable bowel syndrome (IBS) and other digestive disorders. Examples include almond milk, lactose-free milk, rice milk, coconut milk, lactose-free yogurt, and hard cheeses, bananas, blueberries, cantaloupe, grapefruit, honeydew, kiwi, lemon, lime, oranges, strawberries. bamboo shoots, bean sprouts, bok choy, carrots, chives, cucumbers, eggplant, ginger, lettuce, olives, parsnips, potatoes, spring onions, turnips, beef, pork, chicken, fish, eggs, and tofu, almonds, macadamia nuts, peanuts, pine nuts, walnuts, oats, oat bran, rice bran, gluten-free pasta, quinoa, white rice, and corn flour.

[0025] In any embodiment, the edible product is a low pH, heat treated, product. Examples are low pH carbonated drinks and fermented dairy products whose production involves heat treatment such as pasteurisation, UHT or sterilisation. The term “low pH” ideally means a pH of 2-5, preferably 2-4 or 2.5 to 3.5. The term “heat treated” means subject to a commercial heat treatment process such as pasteurisation, UHT or sterilisation.

[0026] In any embodiment, the composition is a unit dose product comprising a multiplicity of microencapsulates, in which the microencapsulates in the unit dose product comprises 1-20g of prebiotic dietary fibre.

[0027] In a third aspect, there is provided a method of preventing or inhibiting gastrointestinal gas formation or bloating associated with consumption of a FODMAP in a subject, comprising administering microencapsulates according to the disclosure to the subject.

[0028] In any embodiment, the subject has Inflammatory Bowel Syndrome (IBS).

[0029] In a fourth aspect, there is provided a method of preventing or inhibiting diarrhoea or constipation associated with consumption of a FODMAP in a subject, comprising administering microencapsulates according to the disclosure to the subject. In any embodiment, the subject has a sensitive gut.

[0030] In any embodiment, the subject has small intestinal bacterial overgrowth (SIBO).

[0031] In any embodiment, the subject has FODMAPs intolerance.

[0032] In any embodiment, the method comprises administering a dose of 1 -30g, 1 -25g, 1- 20g, 1 -15g, 5-30g, 5-25g, 5-20g, 5-15g,10-30g, 10-25g, 10-20g, 10-15g of microencapsulate to the subject.

[0033] In any embodiment, the low glycaemic index (Gl) oligosaccharide comprises or consists of a prebiotic dietary fibre.

[0034] In any embodiment, the protein is denatured protein. In any embodiment, the protein comprises or consists of plant or dairy protein.

[0035] In any embodiment, the method comprises administering the dose of microencapsulate to the subject every day, every two days, every three days, once or twice a week, or once or twice a month.

[0036] In a fifth aspect, there is provided a method of supplementing a diet of a subject on a low FODMAP diet comprising administering microencapsulates according to the disclosure to the subject. The term “low FODMAP diet” means a diet that is deficient in High FODMAP foods such as fruits (including apples, mangos, pears, watermelon), honey, high-fructose com syrup, agave, milk from cows, goats, or sheep), custard, yogurt, ice cream, Rye and Wheat, asparagus, broccoli, cabbage, onions, garlic, Legumes, such as beans (including baked beans), lentils, chickpeas, and soybeans and sugar alcohols and fruits that have pits or seeds, such as apples, apricots, avocados, cherries, figs, peaches, pears, or plums. Other aspects and preferred embodiments of the invention are defined and described in the other claims set out below.

[0037] Detailed Description of the Invention

[0038] All publications, patents, patent applications and other references mentioned herein are hereby incorporated by reference in their entireties for all purposes as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference and the content thereof recited in full.

[0039] Definitions and general preferences

[0040] Where used herein and unless specifically indicated otherwise, the following terms are intended to have the following meanings in addition to any broader (or narrower) meanings the terms might enjoy in the art:

[0041] Unless otherwise required by context, the use herein of the singular is to be read to include the plural and vice versa. The term "a" or "an" used in relation to an entity is to be read to refer to one or more of that entity. As such, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein.

[0042] As used herein, the term "comprise," or variations thereof such as "comprises" or "comprising," are to be read to indicate the inclusion of any recited integer (e.g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g. features, element, characteristics, properties, method / process steps or limitations) but not the exclusion of any other integer or group of integers. Thus, as used herein the term "comprising" is inclusive or open- ended and does not exclude additional, unrecited integers or method / process steps. As used herein, the term “disease” is used to define any abnormal condition that impairs physiological function and is associated with specific symptoms. The term is used broadly to encompass any disorder, illness, abnormality, pathology, sickness, condition or syndrome in which physiological function is impaired irrespective of the nature of the aetiology (or indeed whether the aetiological basis for the disease is established). It therefore encompasses conditions arising from infection, trauma, injury, surgery, radiological ablation, poisoning or nutritional deficiencies.

[0043] As used herein, the term "treatment" or "treating" refers to an intervention (e.g., the administration of an agent to a subject) which cures, ameliorates or lessens the symptoms of a disease or removes (or lessens the impact of) its cause(s). In this case, the term is used synonymously with the term “therapy”.

[0044] Additionally, the terms "treatment" or "treating" refers to an intervention (e.g., the administration of an agent to a subject) which prevents or delays the onset or progression of a disease or reduces (or eradicates) its incidence within a treated population. In this case, the term treatment is used synonymously with the term “prophylaxis”.

[0045] As used herein, an effective amount or a therapeutically effective amount of a compound of the invention defines an amount that can be administered to a subject without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio, but one that is sufficient to provide the desired effect, e.g. the treatment or prophylaxis manifested by a permanent or temporary improvement in the subject's condition. The amount will vary from subject to subject, depending on the age and general condition of the individual, mode of administration and other factors. Thus, while it is not possible to specify an exact effective amount, those skilled in the art will be able to determine an appropriate "effective" amount in any individual case using routine experimentation and background general knowledge. A therapeutic result in this context includes eradication or lessening of symptoms, reduced pain or discomfort, prolonged survival, improved mobility and other markers of clinical improvement. A therapeutic result need not be a complete cure.

[0046] In the context of treatment and effective amounts as defined above, the term subject (which is to be read to include "individual", "animal", "patient" or "mammal" where context permits) defines any subject, particularly a mammalian subject, for whom treatment is indicated. Mammalian subjects include, but are not limited to, humans, domestic animals, farm animals, zoo animals, sport animals, pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows; primates such as apes, monkeys, orangutans, and chimpanzees; canids such as dogs and wolves; felids such as cats, lions, and tigers; equids such as horses, donkeys, and zebras; food animals such as cows, pigs, and sheep; ungulates such as deer and giraffes; and rodents such as mice, rats, hamsters and guinea pigs. In preferred embodiments, the subject is a human.

[0047] “FODMAP” refers to short-chain carbohydrates that are poorly absorbed in the small intestine and ferment in the colon - the term is an acronym for “Fermentable Oligosaccharides, Disaccharides, Monosaccharides, And Polyols”. They include short-chain oligosaccharide polymers of fructose (fructans) and galactooligosaccharides (GOS, stachyose, raffinose), disaccharides (e.g. melibiose, cellobiose), monosaccharides (e.g. tagatose, allulose), and sugar alcohols (polyols), such as sorbitol, mannitol, xylitol, and maltitol. Most FODMAPs are naturally present in food and the human diet, but the polyols may be added artificially in commercially prepared foods and beverages. In any embodiment, the FODMAP is selected from short-chain oligosaccharide polymers of fructose (fructans) and galactooligosaccharides (GOS, stachyose, raffinose), melibiose, cellobiose, tagatose, allulose, and sugar alcohols (polyols), such as sorbitol, mannitol, xylitol, and maltitol.

[0048] “Microencapsulate”: should be understood to mean particles comprising gelled polymer for example denatured protein for example denatured pea protein and typically having an average diameter of 50 to 500 microns as determined using a method of laser diffractometry described in Doherty et al. (Development and characterisation of whey protein micro-beads as potential matrices for probiotic protection, S.B. Doherty, V.L. Gee, R.P. Ross, C. Stanton, G.F. Fitzgerald, A. Brodkorb, Food Hydrocolloids Volume 25, Issue 6, August 2011 , Pages 1604- 1617). The microencapsulate may have a core-shell morphology (e.g. a core comprising FODMAP and a shell comprising gelled polymer such as denatured protein) or a multi-nuclear morphology (continuous polymer matrix with pockets of FODMAP dispersed throughout the matrix). Microencapsulates of active agents, and methods for their production, are described in the literature, including Doherty et al. The microencapsulates may be formed by a 2- or 3- nozzle stray-drying method described in WO2023075688A2, by extrusion and cold gelation methods (single or double nozzle) described in PCT / EP2010 / 054846, PCT / EP2014 / 062154, US 20170367373, US2021 / 0100749, or by a fluidised bed method described in WO2016096931 . In any embodiment, the microencapsulate comprises greater than 30%, 40%, 50%, 60%, 70% or 80% FODMAP (w / w). In any embodiment, the microencapsulate comprises 30% to 80% FODMAP and 20% to 70% polymer (e.g. denatured protein)(w / w). In any embodiment, the microencapsulate comprises 50% to 80% FODMAP and 20% to 50% polymer (e.g. denatured protein)(w / w). In any embodiment, the microencapsulate comprises 60% to 80% FODMAP and 20% to 40% polymer (e.g. denatured protein)(w / w). In any embodiment, the microencapsulate comprises a carrier material, for example a sugar or protein. In any embodiment, the microencapsulate comprises 5% to 20% carrier material. In any embodiment, the microencapsulates do not contain probiotic bacteria. In any embodiment, the microencapsulates have an average dimension of 50 - 1000, 50 - 500, 50 - 400, 50 - 300, 50 - 200, 100 - 500, 100 - 300 pm.

[0049] “Polymer” refers to a protein or a polysaccharide. The protein is generally denatured and may be a dairy protein, for example whey protein in any form (e.g. an isolate or concentrate), milk protein, a plant protein for example pea or soy protein. The polysaccharide may be an alginate or gelatin. The polymer should be able to gel (crosslink) in suitable conditions, e.g. by heating, by immersion in a suitable polymerisation bath (e.g. an acidic bath), or by reaction with a crosslinking agent under suitable conditions. Polymerisation baths may contain buffered acetic acid or citric acid.

[0050] “Gastro-resistant”: means that the microencapsulates can survive intact for at least 60 minutes in the simulated stomach digestion model described in Minekus et al., 1999 and 2014 (A computer-controlled system to simulate conditions of the large intestine with peristaltic mixing, water absorption and absorption of fermentation product, Minekus, M., Smeets-Peeters M, Bernalier A, Marol-Bonnin S, Havenaar R, Marteau P, Alric M, Fonty G, Huis in't Veld JH, Applied Microbiology Biotechnology. 1999 Dec; 53 (1 ): 108-14) and (Minekus et al., 2014, A standardised static in vitro digestion method suitable for food - an international consensus, Minekus, A. et al., Food Function, 2014, 5, 1113).

[0051] “Ileal-sensitive”: means that the microencapsulates are capable of releasing their contents in vivo in the ileum of a mammal.

[0052] “Cold-gelated”: means formed by cold-gelation, in which liquid microdroplets are extruded or sprayed into a gelling bath and immediately cured in a gelling bath due to polymerization of the denatured protein surface film. The bath may be heated or sold. Examples of cold-gelation are described in the literature, for example PCT / EP2010 / 054846 and PCT / EP2014 / 062154.

[0053] “Unit dose” refers to a product containing a defined amount of microencapsulate, for example a sachet of microencapsulate powder, a pill, a capsule, a lozenge, a beverage, or a food product. Typically, the unit dose contains 1 -10g, 1-9g, 1 -8g, 1 - 6g, 1 -5g, 1 -3g or 1 -2g, 5-10g, 7-10g, or 8-10g, 2-8g, 3-7g or 5-6g of microencapsulated prebiotic dietary fibre.

[0054] “Slow release” means that the FODMAP is released slowly as the microencapsulates pass along the gut. Typically, the period of release is at least 1 , 2, 3, 4, 5, 6 or 7 hours. Typically, the period of release is 1-6, 2-6, 3-6, 4-6, 1-5, 2-5, 3-5, 1-4, 1-3, 1-2 or 2-4 hours. The microencapsulates described herein are generally slow-release microencapsulates.

[0055] Exemplification

[0056] The invention will now be described with reference to specific Examples. These are merely exemplary and for illustrative purposes only: they are not intended to be limiting in any way to the scope of the monopoly claimed or to the invention described. These examples constitute the best mode currently contemplated for practicing the invention.

[0057] Example 1

[0058] Microencapsulates were made according to the cold gelation method described in US2021 / 0100749 (Method 1 ) with the exception that sucrose was replaced with FOS. The resultant microencapsulates have a core-shell morphology with a core of FOS encapsulated within a shell of denatured whey protein.

[0059] Example 2

[0060] Microencapsulates were made according to the fluidised bed method described in US2021 / 0100749 (Method 2) with the exception that sucrose was replaced with FOS. The resultant microencapsulates have a core-shell morphology with a core of FOS encapsulated within a shell of denatured whey protein.

[0061] Example 3

[0062] Microencapsulates were made according to the cold gelation method described in Example 1 with the exception that the sucrose was replaced with GOS and the denatured whey protein was replaced with denatured pea protein prepared according to the methods described in US20170367373. The resultant microencapsulates have a core-shell morphology with a core of GOS encapsulated within a shell of denatured pea protein.

[0063] Example 4 Microencapsulates were made according to the cold gelation method described in LIS20170367373 (Examples 3) in which the denatured pea protein solution made according to Example 2 is combined with FOS. The resultant microencapsulates have a multinuclear morphology with a continuous matrix of denatured pea protein and pockets of FOS distributed throughout the matrix.

[0064] Example 5

[0065] A sachet of microencapsulates (15g) prepared according to Example 2 was added to a 100ml of beverage. The sachet contained approximately 8g of FOS. The sachet was consumed by a subject and the subject was monitored for flatulence and feelings of bloating for a period of 5 hours after consuming the beverage. As a control, 8g of non-encapsulated FOS was administered to a control subject in 100ml of the same beverage. The level of flatulence and feeling of bloating experienced by the control subject was significantly greater than the test subject.

[0066] Example 6

[0067] The slow-release properties of microencapsulated FOS were investigated. A sachet of microencapsulates (15g) prepared according to Example 2 was added to a pig feed containing no prebiotic fibre. The sachet contained approximately 8g of FOS. The supplemented pig feed was fed to a pig and a sample of the contents of the pigs ileum was obtained every hour over a 6 hour period and analysed for microencapsulate breakdown. The analysis indicates that the microencapsulates remain intact after passage through the stomach and break down slowly in the ileum, with no evidence of fast bolus release of FOS.

[0068] Example 7

[0069] Microencapsulates were made according to the cold gelation method described in Example 1 with the exception that the sucrose was replaced with tagatose and the denatured whey protein was replaced with denatured pea protein prepared according to the methods described in US20170367373. The resultant microencapsulates have a core-shell morphology with a core of tagatose encapsulated within a shell of denatured pea protein. Example 8

[0070] Microencapsulates were made according to the cold gelation method described in Example 1 with the exception that the sucrose was replaced with mellibiose and the denatured whey protein was replaced with denatured pea protein prepared according to the methods described in US20170367373. The resultant microencapsulates have a core-shell morphology with a core of mellibiose encapsulated within a shell of denatured pea protein. Equivalents

[0071] The foregoing description details presently preferred embodiments of the present invention. Numerous modifications and variations in practice thereof are expected to occur to those skilled in the art upon consideration of these descriptions. Those modifications and variations are intended to be encompassed within the claims appended hereto.

Claims

CLAIMS:

1. A microencapsulate comprising a FODMAP encapsulated within a protein matrix, in which the microencapsulate is gastric-resistant and ileal-sensitive.

2. A microencapsulate according to Claim 1 , having a core-shell morphology, in which the shell comprises the protein matrix.

3. A microencapsulate according to Claim 1 or 2, in which the protein matrix is configured to release the FODMAP in the ileum over a period of 1 to 6 hours.

4. A microencapsulate according to any preceding Claim, in which the protein matrix comprises denatured plant or dairy protein.

5. A microencapsulate according to any preceding Claim, in which the FODMAP comprises fructooligosaccharide (FOS) and / or galactooligosaccharides (GOS).

6. A microencapsulate according to any preceding Claim, having a core-shell morphology, in which the core consists essentially of FODMAP.

7. A microencapsulate according to any preceding Claim, having a dimension of 50 to 500 pm.

8. A microencapsulate according to any preceding Claim, consisting essentially of FODMAP and a protein matrix.

9. A microencapsulate according to any preceding Claim, that does not contain any bacteria.

10. A composition comprising a multiplicity of microencapsulates according to any preceding Claim, in which the composition is selected from a powder, a food product, a beverage, and a nutritional supplement.11 . A composition according to Claim 10, which is a low pH, heat-treated, product.

12. Microencapsulates according to any of Claims 1 to 9 for use in a method of preventing or inhibiting gastrointestinal gas formation or bloating associated with consumption of FODMAPs in a subject, in which the method comprises administering a therapeutically effective amount of the microencapsulates to the subject.

13. Microencapsulates according to any of Claims 1 to 9, for use of claim 12, in which the microencapsulates consist essentially of a protein matrix and FODMAP.

14. Microencapsulates according to any of Claims 1 to 9, for use of claim 12 or 13, in which in which the subject suffers from a sensitive gut or Inflammatory Bowel Syndrome.

15. Microencapsulates according to any of Claims 1 to 9, for use in a method of supplementing a diet of a subject on a low FODMAP diet, comprising administering microencapsulates according to any of Claims 1 to 8 to the subject as a supplement to the low FODMAP diet.