Methods of reducing food cravings and related compositions

Abstract

Provided are methods of reducing food cravings in a subject in need thereof. In certain embodiments, the methods comprise administering to the subject a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila in an amount effective to reduce food cravings in the subject. In some instances, the methods comprise administering to the subject a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis in an amount effective to reduce food cravings in the subject. According to some embodiments, the subject experiences from a specific food craving specific for a particular group of foods selected from sweets, carbohydrates and starch, fast food fats, and high-fat foods and any combination thereof. Also provided are oral formulations consisting of Akkermansia muciniphila that reduce food cravings.

Description

[0001] METHODS OF REDUCING FOOD CRAVINGS AND RELATED COMPOSITIONS

[0002] SUMMARY

[0003] Provided are methods of reducing food cravings in a subject in need thereof. In certain embodiments, the methods comprise administering to the subject a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila in an amount effective to reduce food cravings in the subject. In some instances, the methods comprise administering to the subject a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis in an amount effective to reduce food cravings in the subject. According to some embodiments, the subject does not have an eating order or an eating-related disorder. In certain embodiments, the food craving is a craving for a specific food group selected from sweets, carbohydrates and starch, fast food facts, high-fat foods, or a sub-combination thereof. In some instances, the subject is diagnosed as having an eating disorder and the food cravings is a symptom associated with the eating disorder, e.g., Bulimia Nervosa, Binge Eating Disorder (BED), Other Specified Feeding or Eating Disorders (OSFED), Night Eating Syndrome, pica, rumination disorder, anorexia nervosa, binge-eating disorder, avoidant restrictive food intake disorder, overweight-related disorder, obesity-related disorders or any other eating disorder (e.g., compulsive eating disorder) and combinations thereof. In some instances, the subject is identified or has been identified as needing reducing food cravings.

[0004] BRIEF DESCRIPTION OF THE FIGURES

[0005] FIG. 1 : Data demonstrating that Akkermansia muciniphila administration for 3 months reduces overall food cravings.

[0006] FIG. 2: Data demonstrating that Akkermansia muciniphila administration for 3 months reduces overall food cravings.

[0007] FIG. 3A-3D: Data demonstrating that Akkermansia muciniphila administration for 3 months reduces specific food cravings, e.g., carbohydrates and starch, fast food fats, high fats, and sweets, respectively.

[0008] FIG. 4A-4B: (4A): Data showing mean-change (+ / - 28SE) in food cravings from baseline to six weeks upon administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (4B): A bar graph showing reported pills per week over the past month.

[0009] FIG. 5A-5E: Data demonstrating a reduction in total food cravings after six weeks of administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (5A): Change in food craving inventory score. (5B): Food score by timepoint and baseline food score bin. (5C): Food craving sub score from baseline to six weeks across 4 food categories consisting of sweets, carbs / starches, fasts food fat, and high fat. (5D): Data demonstrating that subjects experiencing higher total cravings at baseline exhibited a greater reduction of cravings at six weeks. (5E) A table showing exemplary foods within the 4 food categories.

[0010] FIG. 6: Binning by baseline total cravings value and change in cravings between baseline and six weeks of administration.

[0011] FIG. 7: Data demonstrating that higher subdomain food cravings were reduced from baseline to six weeks.

[0012] FIG. 8: Binning by subdomain baseline values, p values and % improvements.

[0013] FIG. 9A-9B: (9A): Data showing mean-change (+ / - 2*SE) in food cravings from baseline to twelve weeks upon administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (9B): A bar graph showing reported pills per week over the past month.

[0014] FIG. 10A-10D: Data demonstrating a reduction in total food cravings after twelve weeks of administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (10A): Change in food craving inventory score. (10B): Food score by timepoint and baseline food score bin. (10C): Food craving sub score from baseline to twelve weeks across 4 food categories consisting of sweets, carbs / starches, fasts food fat, and high fat. (5D): Data demonstrating that subjects experiencing higher total cravings at baseline exhibited a greater reduction of cravings at twelve weeks.

[0015] FIG. 11A-11 B: (11 A): Data showing mean-change (+ / - 2*SE) in food cravings from baseline to six weeks and then up to twelve weeks upon administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (11 B): A bar graph showing reported pills per week over the past month.

[0016] FIG. 12A-12C: Data demonstrating a reduction in total food cravings after six weeks and up to twelve weeks of administration of a composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. (12A): Change in food craving inventory score. (12B): Food score by timepoint and baseline food score bin. (12C): Food craving sub score from baseline to six weeks and up to twelve weeks across 4 food categories consisting of sweets, carbs / starches, fasts food fat, and high fat. Supplemental figures, tables and information are provided in the Appendix to the specification filed herewith, the disclosure of which is incorporated herein by reference in its entirety.

[0017] DETAILED DESCRIPTION

[0018] Before the methods and compositions of the present disclosure are described in greater detail, it is to be understood that the methods and compositions are not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the methods and compositions will be limited only by the appended claims.

[0019] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the methods and compositions. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the methods and compositions, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the methods and compositions.

[0020] Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.

[0021] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the methods and compositions belong. Although any methods and compositions similar or equivalent to those described herein can also be used in the practice or testing of the methods and compositions, representative illustrative methods and compositions are now described.

[0022] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the materials and / or methods in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present methods and compositions are not entitled to antedate such publication, as the date of publication provided may be different from the actual publication date which may need to be independently confirmed.

[0023] It is noted that, as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0024] It is appreciated that certain features of the methods and compositions, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the methods and compositions, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed, to the extent that such combinations embrace operable processes and / or compositions. In addition, all sub-combinations listed in the embodiments describing such variables are also specifically embraced by the present methods and compositions and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0025] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present methods. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

[0026] METHODS OF REDUCING FOOD CRAVINGS

[0027] Aspects of the present disclosure include methods of reducing food cravings. Food cravings are defined as an intense desire to consume food or a particular subtype or combination thereof that is difficult to resist. In some embodiments, the subject experiences food cravings that are not associated with an eating disorder or eating-related disorder. In some embodiments, the desire is uncontrollable and is often directed to food that are high in sugar (“sugary” foods), salt, unhealthy fats. In some embodiments, the food craving is for foods that are high in carbohydrates and starches. In some other embodiments, the food craving is for fast-food fats. In some embodiments, the food craving is for food with a high fat content. In further embodiments, the food craving is for sweets. In some embodiments, the food craving is for a combination of one or more of sweets, foods rich in carbohydrates and starches, high- fat foods, and fast-food fats.

[0028] According to some embodiments, the methods comprise administering to the subject a composition comprising a microbial population comprising, consisting essentially of, or consisting of Akkermansia muciniphila, in an amount effective to reduce the food cravings in the subject. In certain instances, the methods comprise administering to the subject a composition comprising a microbial population comprising, consisting essentially of, or consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis in an amount effective to reduce food cravings in the subject.

[0029] In certain embodiments, the subject does not have an eating disorder or an eating- related disorder. In certain embodiments, the subject is or has been identified and / or diagnosed with a need for reducing food cravings. In certain other embodiments, the subject has been identified as suffering from an eating disorder. In certain other embodiments, the subject has been identified as suffering from an eating-related disorder. An eating disorder or eating-related disorder is used interchangeably for the purpose of this disclosure and refers to a disorder directed towards food intake. Non-limiting examples of eating disorder or eating- related disorder include Bulimia Nervosa, Binge Eating Disorder (BED), Other Specified Feeding or Eating Disorders (OSFED), night eating syndrome, pica, rumination disorder, anorexia nervosa, binge-eating disorder, avoidant restrictive food intake disorder, overweight- related disorder, obesity-related disorders and any combination thereof. In some embodiments, an eating-disorder is associated with abnormal eating behaviors that can affect the ability of the body to meet nutritional needs of the subject. Eating disorders can affect the mental and physical health of the subject leading to other disorders.

[0030] In some instances, the eating-related disorder is bulimia nervosa. In certain embodiments, the eating-related disorder is overweight-related disorder or obesity-related disorder. According to some embodiments, the eating-related disorder is overweight-related disorder. In one embodiment, the eating-related disorder is obesity-related disorder. In one embodiment, the eating-related disorder is Binge Eating Disorder (BED). In one embodiment, the eating-related disorder is anorexia. In one embodiment, the eating-related disorder is pica. In one embodiment, the eating-related disorder is rumination disorder. In one embodiment, the eating-related disorder is purging disorder. In one embodiment, the eating-related disorder is night eating syndrome. In one embodiment, the eating-related disorder is avoidant restrictive food intake disorder. In some instances, the subject has been identified as suffering from an eating-related disorder that is bulimia nervosa. In certain embodiments, the subject has been identified as suffering from an eating-related disorder that is an overweight-related disorder or obesity- related disorder. According to some embodiments, the eating-related disorder is overweight- related disorder. In one embodiment, the subject has been identified as suffering from an eating-related disorder that is an obesity-related disorder. In one embodiment, the subject has been identified as suffering from the eating-related disorder that is Binge Eating Disorder (BED). In one embodiment, the subject has been identified as suffering from the eating-related disorder is anorexia. In another embodiment, the subject has been identified as suffering from an eating-related disorder is pica. In one embodiment, the subject has been identified as suffering from the eating-related disorder that is a rumination disorder. In one embodiment, the subject has been identified as suffering from the eating-related disorder is purging disorder. In one embodiment, the subject has been identified as suffering from the eating- related disorder is night eating syndrome. In one embodiment, the subject has been identified as suffering from the eating-related disorder is avoidant restrictive food intake disorder.

[0031] In some embodiments, the subject experiences unwanted food cravings, and wherein the administration of a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila reduces, partially or completely, the unwanted food cravings in the subject. In certain embodiments, the subject experiences unwanted food cravings, and wherein the administration of a composition comprising a microbial population comprising or consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis reduces, partially or completely, the unwanted food cravings in the subject.

[0032] The methods of the present disclosure are based, at least in part, on the demonstration herein of the treatment of a subject experiencing food cravings with a composition consisting of Akkermansia muciniphila, or a composition consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. Also demonstrated herein is that certain specific food cravings, e.g., sweets, carbohydrates and starch, fast food fats, and high-fat foods are reduced in the subject by the administration of a composition consisting of Akkermansia muciniphila, or a composition consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

[0033] In some embodiments, the subject has been diagnosed as suffering from an eating disorder or an eating-related disorder, and wherein the food cravings are a symptom associated with the eating disorder or the eating-related disorder.

[0034] In some embodiments, the subject has been identified as suffering from an eating disorder and wherein the food cravings are a symptom associated with the eating disorder. In some embodiments, the subject has been identified as suffering from an eating- related disorder, and wherein the food cravings are a symptom associated with the eating- related disorder.

[0035] In some embodiments, presented herein is a use for reducing food cravings in a subject, wherein the subject has been identified or diagnosed with a need for reduced food cravings.

[0036] Details regarding embodiments of the methods of the present disclosure will now be described.

[0037] Akkermansia muciniphila is a gram negative, strict anaerobe that can play a role in mucin degradation. Akkermansia muciniphila can serve as a primary fermenter, and in some cases, be combined with any one or more of the secondary fermenters. In certain embodiments, if the composition comprises one or more bacterial strains in addition to the Akkermansia muciniphila, then the Akkermansia muciniphila constitutes at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the bacteria in the composition. According to some embodiments, the composition comprises microbes consisting of Akkermansia muciniphila.

[0038] According to some embodiments, the microbial population comprising Akkermansia muciniphila further comprises one or more additional microbial species selected from the group consisting of Anaerostipes caccae, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Butyrivibrio fibrisolvens, Clostridium acetobutylicum, Clostridium aminophilum, Clostridium beijerinckii, Clostridium butyricum, Clostridium colinum, Clostridium indolis, Clostridium orbiscindens, Enterococcus faecium, Eubacterium hallii, Eubacterium rectale, Faecalibacterium prausnitzii, Fibrobacter succinogenes, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus bulgaricus, Lactobacillus easel, Lactobacillus caucasicus, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus lactis, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Oscillospira guilliermondii, Roseburia cecicola, Roseburia inulinivorans, Ruminococcus flavefaciens, Ruminococcus gnavus, Ruminococcus obeum, Streptococcus cremoris, Streptococcus faecium, Streptococcus infantis, Streptococcus mutans, Streptococcus thermophilus, Anaerofustis stercorihominis, Anaerostipes hadrus, Anaerotruncus colihominis, Clostridium sporogenes, Clostridium tetani, Coprococcus eutactus, Eubacterium cylindroides, Eubacterium dolichum, Eubacteriumventriosum, Roseburia faeccis, Roseburia hominis, or Roseburia intestinalis, and any combination thereof.

[0039] In some instances, provided herein, the methods of reducing food cravings in a subject comprises the method of administering to the subject a composition comprising Bifidobacterium infantis (e.g., available from Dupont Danisco as Bifidobacterium longum subsp. infantis Bi-26 (DGCC1 1473); a Clostridium butyricum obtained by the methods described in the Experimental section herein, and the Akkermansia muciniphila strain as deposited at the American Type Culture Collection (ATCC) under ATCC Accession No. BAA- 835.

[0040] Provided herein are also methods of reducing food cravings in a subject, the method comprising administering to the subject a composition comprising a microbial population of Akkermansia muciniphila strain as deposited at the American Type Culture Collection (ATCC) under ATCC Accession No. BAA-835.

[0041] In some embodiments, the Akkermansia muciniphila is grown in a vegetable-based medium. In some embodiments, the Akkermansia muciniphila is grown in a substantially plantbased culture medium. In some embodiments, the Akkermansia muciniphila is grown in a plant-based culture medium, wherein the plant-based culture medium comprises a plant protein hydrolysate. In some embodiments, the Akkermansia muciniphila is grown in a dairy- free and animal-derived component-free media. Examples of media in which the Akkermansia muciniphila may be grown (e.g., PYGveg media and variations thereof) include those described in WO2016070151 A1 , the disclosure of which is incorporated herein by reference in its entirety for all purposes.

[0042] In some embodiments, the composition administered comprises bacteria that are non- viable. In some other embodiments, the composition administered comprises bacteria that are viable. In some embodiments, the composition administered comprises a microbial population of Akkermansia muciniphila that are viable.

[0043] Viability may be measured using flow cytometry (BD Accuri™ C6). Such flow cytometry measurements include staining the cell mixture and detecting the fluorescence of each cell as it flows by. The staining of the cell mixture can be done using any of the stains that preferentially stain cells, especially live and dead cells, or just dead cells. Examples of such stains include, but are not limited to, Thiazole Orange, Propidium Iodide, 7-AAD, DAPI, SYTO 9, fixable dyes like Zombie Aqua™, Zombie Green™, Zombie NIR™, Zombie Red™, Zombie Violet™, Zombie LIV™, and Zombie Yellow™, or any combinations thereof. In one embodiment, the stain used herein for processes of this disclosure is a combination of thiazole orange and Propidium Iodide.

[0044] In further embodiments, the bacteria of the present disclosure are in a viable, lyophilized state. In some embodiments, the bacteria of the present disclosure are not viable.

[0045] In some instances, one or more of the bacteria in any of the compositions described herein may be in a lyophilized state. In some embodiments, the composition administered comprises a microbial population of Akkermansia muciniphila that are lyophilized. In some embodiments, the composition administered comprises a microbial population of Akkermansia muciniphila that are not pasteurized.

[0046] In certain embodiments, the composition administered comprises Akkermansia muciniphila that are alive / viable. Individual doses of the composition may include, as to any one of the one or more microbial populations (e.g., Akkermansia muciniphila) included in a given dose, between about 1 x106and 1 x1015CFLIs (Colony Forming Units) or AFUs (Active Fluorescent Units) per dose. In some cases, the administration will be at least 1 x106AFUs, 1 x107AFUs of the microbes per dose, at least 1 x108AFUs per dose, at least 1 x109AFUs per dose, at least 1x1010AFUs per dose, at least 1x1011AFUs per dose, at least 1x1012AFUs per dose, at least 1 x1013AFUs per dose, at least 1 x1014AFUs per dose, or more.

[0047] According to some embodiments, the methods comprise administering a composition comprising 45-125 million AFUs of Bifidobacterium infantis per dose, 45-125 million AFUs of Clostridium butyricum per dose, and / or 75-125 million AFUs of Akkermansia muciniphila per dose. In some embodiments, the composition comprises 75-125 million AFUs of Akkermansia muciniphila per dose. For example, in some instances, the composition comprises about 100 million AFUs of Akkermansia muciniphila per dose.

[0048] In certain embodiments, the composition comprises at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 6 mg, at least 7 mg, at least 8 mg, at least 9 mg, at least 10 mg, at least 1 1 mg, at least 12 mg, at least 13mg, at least 14 mg, at least 15 mg, at least 16 mg, at least 17 mg, at least 18 mg, at least 19 mg, or at least 20 mg of active Akkermansia muciniphila per dose. For example, in some instances, the composition comprises at least 5 mg of active Akkermansia muciniphila per dose.

[0049] According to some embodiments, the composition comprises at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 6 mg, at least 7 mg, at least 8 mg, at least 9 mg, at least 10 mg, at least 1 1 mg, at least 12 mg, at least 13mg, at least 14 mg, at least 15 mg, at least 16 mg, at least 17 mg, at least 18 mg, at least 19 mg, or at least 20 mg of active Bifidobacterium infantis per dose. For example, in some instances, the composition comprises about 3 mg (e.g., 3.05 mg) of active Bifidobacterium infantis per dose.

[0050] In some instances, the composition comprises at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 6 mg, at least 7 mg, at least 8 mg, at least 9 mg, at least 10 mg, at least 11 mg, at least 12 mg, at least 13mg, at least 14 mg, at least 15 mg, at least 16 mg, at least 17 mg, at least 18 mg, at least 19 mg, or at least 20 mg of active Clostridium butyricum per dose. For example, in some instances, the composition comprises of at least 8 mg of active Clostridium butyricum per dose.

[0051] In some embodiments, the ratio of Akkermansia muciniphila, Bifidobacterium infantis, Clostridium butyricum in the composition is 1 .2-2.0:0.6-1 .4:2.2-3.0, respectively. In some embodiments, the ratio of Akkermansia muciniphila, Bifidobacterium infantis, Clostridium butyricum in the composition is 1 .6:1 :2.6, respectively.

[0052] When the methods comprise administering to the subject a composition comprising Akkermansia muciniphila, the composition may be administered via any suitable route of administration. Non-limiting examples of suitable routes of administration include oral administration. For example, when the methods comprise administering to the subject a composition comprising Akkermansia muciniphila, in some instances, the administering is by oral administration. In certain embodiments, when administered orally, the composition is in pill form. Non-limiting examples of such formulations include tablets, capsules, or the like. According to some embodiments, when administered orally, the composition is a food, drink, dietary supplement, food supplement, or food additive. In some embodiments, the method comprise administration of a composition in the form of a pill, a capsule, a lozenge, a food bar, or a gummy ball. In certain embodiments, the methods comprise administration of a composition formulated as a liquid composition. In practice, liquid forms adapted for oral administration include, but are not limited to, solutions, suspensions, drinkable solutions, elixirs, sealed phial, potion, drench, syrup, and sprays.

[0053] In some embodiments, the methods comprise administering to the subject a composition formulated such that it comprises a coating that does not fully degrade until it exits the stomach of the subject. The formulation is aimed at delivering the microbial contents of the composition to the ileum and / or colon regions of the subject.

[0054] In some embodiments, such a coating is an enteric-coating. In some embodiments, provided herein is a formulation of composition comprising a microbial population of Akkermansia muciniphila further comprising an enteric-coating that does not degrade until after it exits the stomach of the subject.

[0055] An enteric-coating protects the contents of the oral formulation, for example, pill or capsule, from the acidity of the stomach and provides delivery to the ileum and / or upper colon regions. Non-limiting examples of enteric coatings include pH sensitive polymers (e.g., eudragit FS30D), methyl acrylate-methacrylic acid copolymers, cellulose acetate succinate, hydroxy propyl methyl cellulose phthalate, hydroxy propyl methyl cellulose acetate succinate (e.g., hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), methyl methacrylate-methacrylic acid copolymers, shellac, cellulose acetate trimellitate, sodium alginate, zein, other polymers, fatty acids, waxes, shellac, plastics, and plant fibers. In some embodiments, the enteric coating is formed by a pH sensitive polymer. In some embodiments, the enteric coating is formed by eudragit FS30D.

[0056] The enteric coating can be designed to dissolve at any suitable pH. In some embodiments, the enteric coating is designed to dissolve at a pH greater than about pH 6.5 to about pH 7.0. In some embodiments, the enteric coating is designed to dissolve at a pH greater than about pH 6.5. In some embodiments, the enteric coating is designed to dissolve at a pH greater than about pH 7.0. The enteric coating can be designed to dissolve at a pH greater than about: 5, 5.1 , 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1 , 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1 , 7.2, 7.3, 7.4, or 7.5 pH units.

[0057] In certain embodiments, the composition comprises an Akkermansia muciniphila supernatant or fraction thereof comprising one or more agents that reduce food cravings. By Akkermansia muciniphila “supernatant” is meant a medium in which Akkermansia muciniphila has been cultured. As demonstrated herein, during culture, Akkermansia muciniphila secretes agents known to reduce food cravings into the culture medium. In certain embodiments, a fraction of an Akkermansia muciniphila supernatant comprises, consists essentially of, or consists of, one or more of the one or more agents that reduce food cravings.

[0058] In certain embodiments, the composition consists of an Akkermansia muciniphila supernatant or fraction thereof, comprising one or more agents that reduce food cravings. The agents that reduce food cravings have a direct or indirect effect in inducing GLP-1 (glucagon- like peptide-1 ) levels in the subject. The increase in GLP-1 levels incurs a number of metabolic effects including glucose-dependent stimulation of insulin secretion, decrease of gastric emptying, inhibition of food intake, increase of natriuresis and diuresis, and modulation of rodent p-cell proliferation.

[0059] In certain other embodiments, the composition consists of an Akkermansia muciniphila supernatant or fraction thereof, comprising one or more agents that reduce food cravings have a direct or indirect effect in inducing GLP-2 (Glucagon-like peptide -2), leptin, ghrelin, and / or PYY (peptide tyrosine tyrosine) levels in the subject.

[0060] When the methods comprise administering to the subject a composition comprising an Akkermansia muciniphila supernatant or fraction thereof comprising one or more agents that reduce food cravings, the composition may be administered via any suitable route of administration. Non-limiting examples of suitable routes of administration include oral administration.

[0061] Any of the compositions administered according to the methods of the present disclosure may include additional components, including but not limited to, at least one preservative. In particular embodiments, the composition may contain an effective amount of a preservative. An “effective” amount of a preservative is any amount that preserves or increases the shelf life of the composition beyond what would be obtained if the preservative were not present in the formulation. Examples of such preservatives include, but are not limited to, Vitamin E, Vitamin C, butylatedhydroxyanisole (BHA). butylatedhydroxytoluene (BHT), disodium ethylenediaminetetraacetic acid (EDTA), polyphosphates, citric acid, benzoates, sodium benzoate, sorbates, propionates, and nitrites.

[0062] Formulation of the composition provided herein optionally, further includes the addition of one or more agents to enhance stability and / or survival of the microbial formulation. Nonlimiting example of stabilizing agents include genetic elements, glycerin, ascorbic acid, skim milk, lactose, tween, alginate, xanthan gum, carrageenan gum, mannitol, palm oil, and poly- L-lysine (POPL).

[0063] Any of the compositions administered according to the methods of the present disclosure may include additional components, including but not limited to, at least one excipient. In some preferred embodiments, the excipient used is magnesium stearate.

[0064] Any of the compositions administered according to the methods of the present disclosure may include additional components, including but not limited to, at least one prebiotic. In some embodiments, the composition comprises a prebiotic chosen from the group consisting of inulin, green banana, reishi, tapioca, oats, pectin, potato or extracts thereof, complex carbohydrates, complex sugars, resistant dextrins, resistant starch, amino acids, peptides, nutritional compounds, biotin, polydextrose, fructooligosaccharide (FOS), galactooligosaccharides (GOS), starch, lignin, psyllium, chitin, chitosan, gums (e.g. guar gum), high amylose cornstarch (HAS), cellulose, b-glucans, hemi-celluloses, lactulose, mannooligosaccharides, mannan oligosaccharides (MOS), oligofructose-enriched inulin, oligofructose, oligodextrose, tagatose, trans-galactooligosaccharide, pectin, resistant starch, xylooligosaccharides (XOS), and any combination thereof. In some preferred embodiments, the prebiotic used is inulin.

[0065] A variety of subjects are treatable according to the methods of the present disclosure. The term “subject” as used herein refers to any living organism, including, but not limited to, humans, nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats, rabbits and guinea pigs, and the like. The term does not denote a particular age or sex. In certain embodiments, the subject is a mammal. In some embodiments, the subject is human.

[0066] According to some embodiments, the subject (e.g., a human subject) has an eating disorder. In some instances, prior to the administering, the subject has been identified as having an eating disorder. In certain embodiments, prior to the administering, the subject has been identified as having an eating disorder, and the method is for treating the eating disorder by reducing the food cravings in the subject. According to some embodiments, the eating disorder is bulimia nervosa, Binge Eating Disorder (BED), Other Specified Feeding or Eating Disorders (OSFED), night eating syndrome, or any other compulsive eating disorder and combinations thereof. In some instances, the eating disorder is Bulimia Nervosa. In some instances, the eating disorder is Other Specified Feeding or Eating Disorders (OSFED).

[0067] In certain embodiments, the eating disorder is a psychological disorder. In certain embodiments, the eating disorder is a Binge Eating Disorder (BED). In one embodiment, the eating-related disorder is Bulimia. In one embodiment, the eating-related disorder is overweight-related disorder or obesity-related disorder. In one embodiment, the eating-related disorder is overweight-related disorder. In one embodiment, the eating-related disorder is obesity-related disorder. In one embodiment, the eating-related disorder is anorexia. In one embodiment, the eating-related disorder is pica. In one embodiment, the eating-related disorder is rumination disorder. In one embodiment, the eating-related disorder is purging disorder. In one embodiment, the eating-related disorder is night eating syndrome. In one embodiment, the eating-related disorder is avoidant restrictive food intake disorder.

[0068] According to some embodiments, the subject is a human subject experiencing unwanted food cravings. In some embodiments, the subject is administered a microbial composition comprising Akkermansia muciniphila in an amount effective to reduce the food cravings in the subject.

[0069] In certain embodiments, an effective amount of the composition is administered to reduce one or more of a particular category of food craving. There are at least four recognized categories of food to determine food craving in subjects - sweets (food with high glucose or sugar content; commonly known as “sugary” foods, e.g., cake, cinnamon rolls, ice cream, candy, sweet drinks, chocolate, cookies, donuts, pastry, brownies, etc. ); high fats (food with high saturated fat content or oily foods, e.g., cheese, bacon, fried foods, chips, fried meat, fried fish, etc.); carbohydrates and starches (food with high carbohydrate content or starchy food; e.g., white rice, noodles, cheese, pizza, pasta, potatoes, hamburger etc.); and fast food fats (junk food or fast-food with a high content of unhealthy fats such as pizza, French fries, and burgers).

[0070] In some instances, the effective amount of the composition is administered to a subject in need thereof, for a period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks or more to reduce the overall food cravings by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the overall food cravings in the absence of the administration of the composition.

[0071] By “effective amount” is meant a dosage sufficient to produce a desired result, e.g., an amount sufficient to effect beneficial or desired therapeutic (including prophylactic) results, such as the reduction in overall food cravings or a specific food craving as compared to a control. According to some embodiments, the effective amount reduces overall food cravings or a specific food craving by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the overall food cravings or a specific food craving in the absence of the administration of the composition.

[0072] According to some embodiments, the effective amount reduces overall food cravings by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the overall food cravings in the absence of the administration of the composition.

[0073] In some embodiments, the subject experiences a specific food craving which is a specific craving for sweets, and the administration reduces the specific food craving for sweets by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the specific food craving for sweets in the absence of the administration of the composition.

[0074] In some embodiments, the subject experiences a specific food craving which is a specific craving for carbohydrates and starch, and the administration reduces the specific food craving for carbohydrates and starch by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the specific food craving for carbohydrates and starch in the absence of the administration of the composition.

[0075] In some embodiments, the subject experiences a specific food craving which is a specific craving for fast-food fats, and the administration reduces the specific food craving for fast-food fats by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the specific food craving for fast-food fats in the absence of the administration of the composition.

[0076] In some embodiments, the subject experiences a specific food craving which is a specific craving for high-fat foods, and the administration reduces the specific food craving for high-fat foods by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the specific food craving for high-fat foods in the absence of the administration of the composition.

[0077] In some embodiments, the food cravings are quantified by the FCI-II scores from a scale of 1 -5, as described in White et al. (2002) Obesity Research 10(2):107-1 14 paper - the disclosure of which is incorporated herein by reference in its entirety for all purposes. As taught therein, 1 = never, 2= rarely, 3=sometimes, 4-often, and 5=always / almost every day. Based on these scores, the foods cravings were binned into three groups - mild (scores of 1 -2); moderate (sometimes experiencing cravings, scores of 2-2.5) and high (almost always / everyday, scores of 2.5-5) for purposes of the study disclosed herein.

[0078] In some embodiments described herein, the subject experiences overall food cravings, and the administration reduces the overall food cravings from high to moderate, moderate to mild, or high to mild, as binned according to scores obtained by the scale of White et al. supra).

[0079] In some embodiments described herein, the subject experiences a specific food craving which is a specific craving for sweets, and the administration reduces the specific food craving for sweets from high to moderate, moderate to mild, or high to mild, as binned according to scores obtained by the scale of White et al. supra).

[0080] In some embodiments described herein, the subject experiences a specific food craving which is a specific craving for carbohydrates and starch, and the administration reduces the specific food craving for carbohydrates and starch from high to moderate, moderate to mild, or high to mild, as binned according to scores obtained by the scale of White et al. {supra).

[0081] In some embodiments described herein, the subject experiences a specific food craving which is a specific craving for fast-food fats, and the administration reduces the specific food craving for fast-food fats from high to moderate, moderate to mild, or high to mild, as binned according to scores obtained by the scale of White et al. {supra).

[0082] In some embodiments described herein, the subject experiences a specific food craving which is a specific craving for high-fat foods, and the administration reduces the specific food craving for high-fat foods from high to moderate, moderate to mild, or high to mild, as binned according to scores obtained by the scale of White et al. {supra).

[0083] An effective amount can be administered in one or more administrations. In some cases, the methods comprise administration of multiple doses over a period of time. In certain embodiments, administration may comprise administration of 1 , 2, 3, 4, 5, 6 or more doses over the period of a day. In some cases, such daily administration may occur 1 day, 2 days, 3 days, 4 days, 5 days, 6 days or 7 days during a week. In some cases, such weekly administration may occur over the course of 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 10 weeks, 12 weeks or longer. In some cases, such longer term administration may occur over the course of 1 month, 2 months, 3 months, 4 months, 5, months, 6 months, 7 months, 8 months, 9 months, 10 months, 1 1 months, 12 months or longer. In some cases, administration may be ongoing in order to maintain effects of such treatment, e.g., with the above described administration occurring over the course of 1 year, 2 years, 3 years or longer. For purposes of completeness, the present disclosure is further defined in the following numbered clauses.

[0084] 1 . A method of reducing food cravings in a human subject in need thereof, the method comprising: administering to the human subject a composition comprising a microbial population, wherein the microbial population comprises Akkermansia muciniphila in an amount effective to reduce the food cravings in the human subject.

[0085] 2. The method of clause 1 , wherein the microbial population consists of Akkermansia muciniphila.

[0086] 3. The method of clause 1 , wherein the microbial population comprises Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

[0087] 4. The method of clause 3, wherein the microbial population consists of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

[0088] 5. The method of any one of clauses 1 -4, wherein the food cravings are unwanted food cravings.

[0089] 6. The method of any one of clauses 1 -5, wherein the subject experiences unwanted food cravings, and wherein the administering reduces the unwanted food cravings.

[0090] 7. The method of any one of clauses 1 -6, wherein the food craving is a craving for a specific group of food.

[0091] 8. The method of clause 7, wherein the food craving is a specific craving for sweets.

[0092] 9. The method of clause 7, wherein the food craving is a specific craving for carbohydrates and starch foods.

[0093] 10. The method of clause 7, wherein the food craving is a specific craving for fast food fats.

[0094] 1 1 . The method of clause 7, wherein the food craving is a specific craving for high-fat foods.

[0095] 12. The method of any one of clauses 1 -1 1 , wherein the subject does not have an eating disorder or an eating-related disorder.

[0096] 13. The method of any one of clauses 1 -1 1 , wherein the subject has been diagnosed as suffering from an eating disorder or an eating-related disorder, and wherein the food cravings are a symptom associated with the eating disorder or the eating-related disorder.

[0097] 14. The method of clause 13, wherein the subject has been diagnosed as suffering from an eating disorder or eating-related selected from the group comprising of: Bulimia Nervosa, Binge Eating Disorder (BED), Other Specified Feeding or Eating Disorders (OSFED), night eating syndrome, pica, rumination disorder, anorexia nervosa, binge-eating disorder, avoidant restrictive food intake disorder, overweight-related disorder, obesity-related disorders and any combination thereof.

[0098] 15. The method of any one of clauses 1 -14, wherein the administering is by oral administration.

[0099] 16. The method of clause 15, wherein the composition is in a pill form.

[0100] 17. The method of clause 16, wherein the pill is a tablet or capsule.

[0101] 18. The method of any one of clauses 1 -17, wherein the Akkermansia muciniphila is not pasteurized.

[0102] 19. The method of clause 18, wherein the Akkermansia muciniphila is viable.

[0103] 20. The method of any one of clauses 1 -19, wherein the microbial population comprises

[0104] 75-125 million AFLIs of Akkermansia muciniphila, optionally wherein the microbial population comprises about 100 million AFLIs of Akkermansia muciniphila.

[0105] 21 . The method of any one of clauses 1 -20, wherein the composition is administered for at least 1 .5 months or at least 3 months, optionally wherein composition is administered daily.

[0106] 22. The method of any one of clauses 1 , 3, or 5-21 , wherein the microbial population comprises one or more additional microbial species selected from the group consisting of; Anaerostipes caccae, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum, Butyrivibrio fibrisolvens, Clostridium acetobutylicum, Clostridium aminophilum, Clostridium beijerinckii, Clostridium colinum, Clostridium indolis, Clostridium orbiscindens, Enterococcus faecium, Eubacterium hallii, Eubacterium rectale,

[0107] Faecal I bacterium prausnitzii, Fibrobacter succinogenes, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus bulgaricus, Lactobacillus easel, Lactobacillus caucasicus, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus lactis, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Oscillospira guilliermondii, Roseburia cecicola, Roseburia inulinivorans, Ruminococcus flavefaciens, Ruminococcus gnavus, Ruminococcus obeum, Streptococcus cremoris, Streptococcus faecium, Streptococcus infantis, Streptococcus mutans, Streptococcus thermophilus, Anaerofustis stercorihominis, Anaerostipes hadrus, Anaerotruncus colihominis, Clostridium sporogenes, Clostridium tetani, Coprococcus eutactus, Eubacterium cylindroides, Eubacterium dolichum, Eubacteriumventriosum, Roseburia faeccis, Roseburia hominis, Roseburia intestinalis, and any combination thereof. 23. The method of any one of clauses 1 -22, wherein the composition further comprises a prebiotic.

[0108] 24. The method of clause 23, wherein the prebiotic is inulin.

[0109] 25. The method of any one of clauses 1 -24, wherein the composition further comprises a preservative.

[0110] 26. The method of any one of clauses 1 -25, wherein the composition further comprises a coating that does not fully degrade until after it exits the stomach of the subject.

[0111] 27. The method of clause 26, wherein the coating is an enteric coating.

[0112] 28. The method of any one of clauses 1 -27, wherein the Akkermansia muciniphila is lyophilized.

[0113] 29. The method of any one of clauses 1 -28, wherein the Akkermansia muciniphila is grown on a vegetable-based-medium.

[0114] 30. The method of any one of clauses 1 -29, wherein the composition is a food, drink, dietary supplement, food supplement, or food additive.

[0115] 31 . A composition comprising a microbial population, wherein the microbial population comprises Akkermansia muciniphila, for use in the method of any one of clauses 1 -30.

[0116] 32. The composition for use of clause 31 , wherein the microbial population consists of Akkermansia muciniphila.

[0117] 33. The composition for use of clause 31 , wherein the microbial population comprises Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

[0118] 34. The composition for use of clause 33, wherein the microbial population consists of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

[0119] The following examples are offered by way of illustration and not by way of limitation.

[0120] EXPERIMENTAL

[0121] Example 1 - Beneficial effects of A. muciniphila on overall food cravings

[0122] In this example, subjects experiencing food cravings were administered an oral composition comprising a microbial population consisting of Akkermansia muciniphila.

[0123] The oral composition comprises between 1 x106and 1x1015CFLIs (Colony Forming Units) or AFUs (Active Fluorescent Units) of the Akkermansia muciniphila, and one or more doses of the oral composition are administered per day. In some embodiments, the composition comprises one million AFUs of Akkermansia muciniphila per pill. The delivery form of the oral composition is an enteric-coated (e.g., pH sensitive polymer) capsule or tablet that does not fully degrade until after it exits from the stomach of the subject. In some embodiments, the oral composition can be administered as a capsule comprising a powdered Akkermansia muciniphila composition.

[0124] For purposes of this example, the participating subjects were administered one capsule per day, wherein the capsule comprised a microbial population consisting of one hundred million AFLIs of Akkermansia muciniphila. Administration of the oral composition comprising Akkermansia muciniphila is scheduled for 3 months. The food cravings in participating subjects are studied using a validated questionnaire called the Food-Craving Inventory II (FCI-I I) (Licensed from the Pennington Biomedical Research Center; White et al. (2002) Obesity Research 10(2):107-114, the disclosure of which is incorporated herein by reference in its entirety for all purposes), which featured a 28-item food list and asked participants to indicate how often they experienced a craving for that food over the past month. (See the full FCI-II in the appendix I). The FCI-I I defined food craving as an intense desire to consume a particular food (or food type) that is difficult to resist.

[0125] Participants indicated the frequency of cravings over the past month for individual food items using a 5-point Likert scale (Never, Rarely, Sometimes, Often, or Always). Foods were grouped into four scales for purposes of this study into: sweets (with foods such as cinnamon rolls and ice cream), high fats (with foods such as bacon and fried chicken), carbohydrates / starches (with foods such as baked potatoes and pasta), and fast-food fats (with foods such as pizza, French fries, and hamburgers).

[0126] The change in participants’ FCI-II scores from baseline (when the participants started taking the oral composition comprising Akkermansia muciniphila) to follow-up (when 3 months were completed) was recorded and analyzed at the end of 3 months of oral administration of the composition.

[0127] The results show, as demonstrated in Fig. 1 , that administration of the composition comprising a microbial population consisting of Akkermansia muciniphila resulted in significant decrease in Food Craving Inventory scores, independent of age and gender (p < 0.001 ). Overall, 91 % of participants experienced reduced overall food cravings.

[0128] Methods for Example 1

[0129] Strain, growth medium and conditions

[0130] A. muciniphila (ATCC Accession No. BAA-835) was grown in a medium having the ingredients and concentrations listed in the following table.

[0131] Supernatant obtention and lyophilization

[0132] A. muciniphila supernatant was collected approximately 16 days post growth under manufacturing conditions in Fed Batch fermentation settings. The OD6oo nm at the time of sample collection was ~ 8 (late exponential to early stationary phase). The culture samples were collected and dispensed into sterile 50mL Eppendorf conical tubes and stored at -80°C. Prior to mass spectrometry analysis, the Akkermansia muciniphila cultures were thawed and filtered-sterilized by a 0.2 pM filter.

[0133] The lyophilization process includes a first cycle at -40°C (5 min at 500mTorr), followed by a primary dry step at 20°C (10 min at 500mTorr), followed by a second cycle at -40°C (999 min at 3000 mTorr).

[0134] Example 2 - Reduction in food cravings is proportional to score at baseline

[0135] In this example, subjects experiencing from food craving are divided into three groups based on their baseline craving scores - (a) Mild category - subjects with baseline score of 1 -2 and experiencing food cravings rarely; (b) Moderate category - subjects with baseline score 2-2.5 and experiencing food cravings sometimes; (c) Higher category - subjects with baseline score 2.5 - 5 and experiencing very high food cravings quite often. None of these categories were limited to subjects experiencing food craving to a particular group of subcategories of foods but was scored for overall food cravings in these subjects. These subjects were administered an oral composition comprising a microbial population consisting of Akkermansia muciniphila.

[0136] The subjects were given an oral composition comprising a microbial population consisting of Akkermansia muciniphila for 3 months at a dosage of one pill daily. Each pill comprised 100 million AFLIs of the Akkermansia muciniphila.

[0137] In one embodiment, the composition of the capsule comprises of Akkermansia muciniphila, inulin, hypermellose and magnesium stearate.

[0138] Data as demonstrated in Fig. 2, groups the results of these three categories based on the scores as shown on the top of each rectangular box. The box on the extreme left is for the mild category (score 1 -2); the middle box is for the moderate category (2- 2.5) and the right most box is for the higher category (score 2.5 - 5).

[0139] The results show that subjects with higher baseline cravings score experienced a greater reduction in their scores at the end of the 3-month study period compared to the other two groups.

[0140] Example 3 - Reduction of specific categories of food cravings by administration of a composition comprising a microbial population consisting of A. muciniphila

[0141] Method: Subjects were provided with a validated questionnaire called the Food- Craving Inventory II and asked participants to indicate how often they experienced a craving for any of the foods listed in the inventory. The foods were categorized as four main groups of food - sweets (with foods such as cinnamon rolls and ice cream), high fats (with foods such as bacon and fried chicken), carbo hydrates / starches (with foods such as baked potatoes and pasta), and fast-food fats (with foods such as pizza, French fries, and hamburgers). See the full FCI-II in appendix I.

[0142] Based on their input, scores were given to the subjects. This score is termed as the baseline score.

[0143] Next, the subjects were administered a composition comprising a microbial population consisting of Akkermansia muciniphila. The subjects were given an oral composition comprising a microbial population consisting of Akkermansia muciniphila for 3 months at a dosage of one pill daily. Each pill comprised a microbial population consisting of 100 million AFLIs of Akkermansia muciniphila.

[0144] At the end of 3 months, the same survey questionnaire was again provided to the participating subjects and scored based on their responses. The scores this time are called follow-up scores.

[0145] Results: A significant reduction of cravings was observed for each of the four specific food categories in the FCI-II survey. That is, food cravings were reduced in all four of the following categories: • Sweets

[0146] • Fast-Food Fats

[0147] • Carbohydrates and starches

[0148] • High Fats

[0149] Scores in the subcategories Fast Food Fats and Sweets showed a slightly greater reduction than scores in the other two subcategories. The results for each food subcategory carbohydrates and starch, fast-food fats, high fats, and sweets, are shown in Figure 3A-3D, respectively.

[0150] Example 4 - Reduction of overall / total and specific food cravings by administration of a composition comprising A. muciniphila, Clostridium butyricum and Bifidobacterium infantis

[0151] In this example, subjects experiencing food cravings were administered an enterically coated oral composition (capsule) comprising a microbial population consisting of Akkermansia muciniphila Clostridium butyricum, and Bifidobacterium infantis. In particular, the microbial population consisted of 6.4 x 10A8 AFLIs of Akkermansia muciniphila, 1.9 x 10A9 AFLIs of Bifidobacterium infantis, and 4.3 x 10A8 AFLIs of Clostridium butyricum per dose. The composition was administered daily for six weeks.

[0152] 274 novel participants completed baseline & six week surveys. Shown in FIG. 4A is mean-change (+ / - 28SE) in food cravings from baseline to six weeks upon administration of the composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. Shown in FIG. 4B is reported pills per week over the past month.

[0153] The food cravings in participating subjects were studied using the validated questionnaire called the Food-Craving Inventory II (FCI-II) (Licensed from the Pennington Biomedical Research Center; White et al. (2002) Obesity Research 10(2):107-1 14), which featured a 28-item food list and asked participants to indicate how often they experienced a craving for that food over the past month, as used in Example 1 above. The FCI-II defined food craving as an intense desire to consume a particular food (or food type) that is difficult to resist.

[0154] The 274 participants indicated the frequency of food cravings over the past 1 .5 months using a 5-point Likert scale (Never (1 ), Rarely (2), Sometimes (3), Often (4), or Always(5)). As shown in FIG. 5A, total food cravings were significantly reduced (p < 0.001 ) after six weeks of daily administration of the composition. Of the 274 participants, 91% experienced reduced overall / total food cravings.

[0155] The 274 participants were divided into groups based on their baseline craving scores - (a) Mild category - subjects with baseline score of 1 -2 and experiencing food cravings rarely; (b) Moderate category - subjects with baseline score 2-2.5 and experiencing food cravings sometimes; (c) Higher category - subjects with baseline score 2.5 - 3; and (d) Highest category - subjects with baseline score 3-5 and experiencing very high food cravings quite often. None of these categories were limited to subjects experiencing food craving to a particular group of subcategories of foods but was scored for overall food cravings. As shown in FIG. 5B, participants with higher total food cravings at baseline experienced a greater reduction of cravings at the six week timepoint, i.e., the reduction in food cravings was proportional to the baseline score.

[0156] The 274 participants indicated the frequency of cravings over the past 1 .5 months for individual food items using the 5-point Likert scale. Foods were grouped into four types / categories for purposes of this study: sweets (with foods such as cinnamon rolls and ice cream), high fats (with foods such as bacon and fried chicken), carbohydrates / starches (with foods such as baked potatoes and pasta), and fast-food fats (with foods such as pizza, French fries, and hamburgers). As shown in FIG. 5C, participants experienced significant reductions (p < 0.001 ) in food cravings across all four categories tested: 88% reduction in cravings for sweets; 87% reduction in cravings for carbs / starches; 85% reduction in cravings for fast food fats; and 82% reduction in cravings for high fat food. Here as well, and as shown in FIG. 5D, higher total cravings at baseline experienced greater reduction of cravings at six weeks. Shown in FIG. 6 is binning by baseline total cravings value and % improvements.

[0157] As shown in FIG. 7, subdomain food cravings were reduced after six weeks of administration of the composition, where subjects experiencing higher subdomain cravings at baseline exhibited greater reduction of subdomain cravings at six weeks. Binning by subdomain baseline values, p values and % improvements is shown in FIG. 8.

[0158] Example 5 - Reduction of overall / total and specific food cravings by administration of a composition comprising A. muciniphila, Clostridium butyricum and Bifidobacterium infantis for 12 weeks

[0159] In this example, as in the previous Example 4, subjects experiencing food cravings were administered an enterically coated oral composition (capsule) comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. In particular, the microbial population consisted of 6.4 x 10A8 AFLIs of Akkermansia muciniphila, 1 .9 x 10A9 AFLIs of Bifidobacterium infantis, and 4.3 x 10A8 AFLIs of Clostridium butyricum per dose. The composition was administered daily for twelve weeks.

[0160] 310 novel participants completed baseline & twelve-week surveys. Shown in FIG. 9A is mean-change (+ / - 2*SE) in food cravings from baseline to twelve weeks upon administration of the composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. Shown in FIG. 9B is reported pills per week over the past month for all 310 participants.

[0161] The food cravings in participating subjects were studied using the validated questionnaire called the Food-Craving Inventory II (FCI-II) (Licensed from the Pennington Biomedical Research Center; White et al. (2002) Obesity Research 10(2):107-1 14), which featured a 28-item food list and asked participants to indicate how often they experienced a craving for that food over the past month, as used in Example 1 above. The FCI-II defined food craving as an intense desire to consume a particular food (or food type) that is difficult to resist.

[0162] The 310 participants indicated the frequency of food cravings over the past 3 months using a 5-point Likert scale (Never (1 ), Rarely (2), Sometimes (3), Often (4), or Always (5)). As shown in FIG. 10A, total food cravings were significantly reduced (p < 0.001 ) after twelve weeks of daily administration of the composition. Of the 310 participants, 93% experienced reduced overall / total food cravings.

[0163] The 310 participants were divided into groups based on their baseline craving scores - (a) Mild category - subjects with baseline score of 1 -2 and experiencing food cravings rarely; (b) Moderate category - subjects with baseline score 2-2.5 and experiencing food cravings sometimes; (c) Higher category - subjects with baseline score 2.5 - 3; and (d) Highest category - subjects with baseline score 3-5 and experiencing very high food cravings quite often. None of these categories were limited to subjects experiencing food craving to a particular group of subcategories of foods but was scored for overall food cravings. As shown in FIG. 10B, participants with higher total food cravings at baseline experienced a greater reduction of cravings at the twelve-week timepoint, i.e., the reduction in food cravings was proportional to the baseline score.

[0164] The 310 participants indicated the frequency of cravings over the past 3 months for individual food items using the 5-point Likert scale. Foods were grouped into four types / categories for purposes of this study: sweets (with foods such as cinnamon rolls and ice cream), high fats (with foods such as bacon and fried chicken), carbohydrates / starches (with foods such as baked potatoes and pasta), and fast-food fats (with foods such as pizza, French fries, and hamburgers). As shown in FIG. 10C, participants experienced significant reductions (p < 0.001 ) in food cravings across all four categories tested: 88% reduction in cravings for sweets; 88% reduction in cravings for carbs / starches; 87% reduction in cravings for fast food fats; and 80% reduction in cravings for high fat food. Here as well, and as shown in FIG. 10D, higher total cravings at baseline experienced greater reduction of cravings at six weeks. Example 6 - Reduction of overall / total and specific food cravings by administration of a composition comprising A. muciniphila, Clostridium butyricum and Bifidobacterium infantis for 6 weeks and 12 weeks

[0165] In this example, as in the previous Examples 4 and 5, subjects experiencing food cravings were administered an enterically coated oral composition (capsule) comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. In particular, the microbial population consisted of 6.4 x 10A8 AFUs of Akkermansia muciniphila, 1.9 x 10A9 AFUs of Bifidobacterium infantis, and 4.3 x 10A8 AFUs of Clostridium butyricum per dose. The composition was administered daily for six weeks and then continued upto twelve weeks.

[0166] 222 novel participants completed baseline, six-week & twelve-week surveys. Shown in FIG. 11 A is mean-change (+ / - 2*SE) in food cravings from baseline to six weeks and twelve weeks, respectively, upon administration of the composition comprising a microbial population consisting of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis. Shown in FIG. 1 1 B is reported pills per week over the past month for all 222 participants.

[0167] The food cravings in participating subjects were studied using the validated questionnaire called the Food-Craving Inventory II (FCI-II) (Licensed from the Pennington Biomedical Research Center; White et al. (2002) Obesity Research 10(2):107-1 14), which featured a 28-item food list and asked participants to indicate how often they experienced a craving for that food over the past month, as used in Example 1 above. The FCI-II defined food craving as an intense desire to consume a particular food (or food type) that is difficult to resist.

[0168] The 222 participants indicated the frequency of food cravings over the past 1 .5 months and 3 months using a 5-point Likert scale (Never (1 ), Rarely (2), Sometimes (3), Often (4), or Always (5)). As shown in FIG. 12A, total food cravings were significantly reduced (p < 0.001 ) after twelve weeks of daily administration of the composition. Of the 222 participants, 90% experienced reduced overall / total food cravings at 6 weeks while 93% experienced reduced overall / total food cravings at 12 weeks.

[0169] The 222 participants were divided into groups based on their baseline craving scores - (a) Mild category - subjects with baseline score of 1 -2 and experiencing food cravings rarely; (b) Moderate category - subjects with baseline score 2-2.5 and experiencing food cravings sometimes; (c) Higher category - subjects with baseline score 2.5 - 3; and (d) Highest category - subjects with baseline score 3-5 and experiencing very high food cravings quite often. None of these categories were limited to subjects experiencing food craving to a particular group of subcategories of foods but was scored for overall food cravings. As shown in FIG. 12B, participants with higher total food cravings at baseline experienced a greater reduction of cravings at the twelve-week timepoint, i.e., the reduction in food cravings was proportional to the baseline score.

[0170] The 222 participants indicated the frequency of cravings over the past, 1 .5 months continued upto 3 months, for individual food items using the 5-point Likert scale. Foods were grouped into four types / categories for purposes of this study: sweets (with foods such as cinnamon rolls and ice cream), high fats (with foods such as bacon and fried chicken), carbohydrates / starches (with foods such as baked potatoes and pasta), and fast-food fats (with foods such as pizza, French fries, and hamburgers). As shown in FIG. 12C, participants experienced significant reductions (p < 0.001 ) in food cravings across all four categories tested: all 222 participants showed sustained improvements by showing reduced food cravings over the 6 week and 12 weeks periods across all four food categories, as shown. Further, it was also observed as in the Examples 4 and 5 above, participants with higher total cravings at baseline experienced greater reduction of cravings at six weeks and consecutively at twelve weeks.

[0171] Methods for Examples 4, 5, and 6

[0172] Akkermansia muciniphila (ATCC Accession No. BAA-835) was prepared as described for Example 1 .

[0173] For preparation of Clostridium butyricum (CBUT), the fermentation process begins with the resuspension of CBUT powder seed stock in media, followed by streak plating onto agar plates. After 24 hours of incubation at 37°C, colonies are inoculated into tubes of media. Incubation at 37°C ensues, during which the optical density (OD) is monitored. When one of the tubes reaches an OD between 0.7-1.0, the culture is aseptically transferred to a bottle containing sterile media. The growth is monitored until OD is between 0.7 and 1.0. The entire culture is aseptically transferred to a bioreactor and further incubated at 37°C. Growth is monitored, and the harvest centrifuge process involves transferring the microbial culture from the fermentor to a centrifuge. Homogenous biomass accumulates within the centrifuge, cryopreservative is added, and is discharged into a collection vessel. Discharge is collected, weighed, and placed in liquid nitrogen, then stored until lyophilization.

[0174] Bifidobacterium infantis was obtained from Dupont Danisco as Bifidobacterium longum subsp. infantis Bi-26 (DGCC1 1473).

[0175] Supplemental figures, tables and information are provided in the Appendix to the Specification filed herewith, the disclosure of which is incorporated herein by reference in its entirety. Accordingly, the preceding merely illustrates the principles of the present disclosure. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein.

Claims

WHAT IS CLAIMED IS:1 . A method of reducing food cravings in a human subject in need thereof, the method comprising: administering to the human subject a composition comprising a microbial population, wherein the microbial population comprises Akkermansia muciniphila in an amount effective to reduce the food cravings in the human subject.

2. The method of claim 1 , wherein the microbial population consists of Akkermansia muciniphila.

3. The method of claim 1 , wherein the microbial population comprises Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

4. The method of claim 3, wherein the microbial population consists of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

5. The method of any one of claims 1-4, wherein the food cravings are unwanted food cravings.

6. The method of any one of claims 1-5, wherein the subject experiences unwanted food cravings, and wherein the administering reduces the unwanted food cravings.

7. The method of any one of claims 1-6, wherein the food craving is a craving for a specific group of food.

8. The method of claim 7, wherein the food craving is a specific craving for sweets.

9. The method of claim 7, wherein the food craving is a specific craving for carbohydrates and starch foods.

10. The method of claim 7, wherein the food craving is a specific craving for fast food fats.1 1 . The method of claim 7, wherein the food craving is a specific craving for high-fat foods.

12. The method of any one of claims 1-11 , wherein the subject does not have an eating disorder or an eating-related disorder.

13. The method of any one of claims 1-11 , wherein the subject has been diagnosed as suffering from an eating disorder or an eating-related disorder, and wherein the food cravings are a symptom associated with the eating disorder or the eating-related disorder.

14. The method of claim 13, wherein the subject has been diagnosed as suffering from an eating disorder or eating-related selected from the group comprising of: Bulimia Nervosa, Binge Eating Disorder (BED), Other Specified Feeding or Eating Disorders (OSFED), night eating syndrome, pica, rumination disorder, anorexia nervosa, binge-eating disorder, avoidant restrictive food intake disorder, overweight-related disorder, obesity-related disorders and any combination thereof.

15. The method of any one of claims 1-14, wherein the administering is by oral administration.

16. The method of claim 15, wherein the composition is in a pill form.

17. The method of claim 16, wherein the pill is a tablet or capsule.

18. The method of any one of claims 1-17, wherein the Akkermansia muciniphila is not pasteurized.

19. The method of claim 18, wherein the Akkermansia muciniphila is viable.

20. The method of any one of claims 1-19, wherein the microbial population comprises 75-125 million AFUs of Akkermansia muciniphila, optionally wherein the microbial population comprises about 100 million AFUs of Akkermansia muciniphila.21 . The method of any one of claims 1-20, wherein the composition is administered for at least 1 .5 months or at least 3 months, optionally wherein composition is administered daily.

22. The method of any one of claims 1 , 3, or 5-21 , wherein the microbial population comprises one or more additional microbial species selected from the group consisting of: Anaerostipes caccae, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum, Butyrivibrio fibrisolvens, Clostridium acetobutylicum, Clostridium aminophilum, Clostridium beijerinckii, Clostridium colinum, Clostridium indolis, Clostridium orbiscindens, Enterococcus faecium, Eubacterium hallii, Eubacterium rectale, Faecalibacterium prausnitzii, Fibrobacter succinogenes, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus caucasicus, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus lactis, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Oscillospira guilliermondii, Roseburia cecicola, Roseburia inulinivorans, Ruminococcus flavefaciens, Ruminococcus gnavus, Ruminococcus obeum, Streptococcus cremoris, Streptococcus faecium, Streptococcus infantis, Streptococcus mutans, Streptococcus thermophilus, Anaerofustis stercorihominis, Anaerostipes hadrus, Anaerotruncus colihominis, Clostridium sporogenes, Clostridium tetani, Coprococcus eutactus, Eubacterium cylindroides, Eubacterium dolichum, Eubacteriumventriosum, Roseburia faeccis, Roseburia hominis, Roseburia intestinalis, and any combination thereof.

23. The method of any one of claims 1-22, wherein the composition further comprises a prebiotic.

24. The method of claim 23, wherein the prebiotic is inulin.

25. The method of any one of claims 1-24, wherein the composition further comprises a preservative.

26. The method of any one of claims 1-25, wherein the composition further comprises a coating that does not fully degrade until after it exits the stomach of the subject.

27. The method of claim 26, wherein the coating is an enteric coating.

28. The method of any one of claims 1-27, wherein the Akkermansia muciniphila is lyophilized.

29. The method of any one of claims 1-28, wherein the Akkermansia muciniphila is grown on a vegetable-based-medium.

30. The method of any one of claims 1-29, wherein the composition is a food, drink, dietary supplement, food supplement, or food additive.31 . A composition comprising a microbial population, wherein the microbial population comprises Akkermansia muciniphila, for use in the method of any one of claims 1 -30.

32. The composition for use of claim 31 , wherein the microbial population consists ofAkkermansia muciniphila.

33. The composition for use of claim 31 , wherein the microbial population comprisesAkkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

34. The composition for use of claim 33, wherein the microbial population consists of Akkermansia muciniphila, Clostridium butyricum, and Bifidobacterium infantis.

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