BE1028330B1 - Composition comprising Akkermansia muciniphila and camu camu extract - Google Patents

Abstract

Disclosed are compositions comprising Akkermansia muciniphila and camu camu extract. The composition is useful for the treatment of prediabetes, type 1 diabetes or type 2 diabetes. A method of making the composition and its use for controlling the blood glucose level of a subject in need thereof are also disclosed.

Description

FIELD OF THE INVENTION The present invention relates to a composition comprising Akkermansia muciniphila and a camu camu extract. In particular, the present invention relates to a composition comprising Akkermansia muciniphila and an extract of camu camu, in which Akkermansia muciniphila is pasteurized. The compositions of the present invention are useful in the prevention or treatment of a metabolic disorder selected from the group comprising obesity, metabolic syndrome, disorders related to insulin deficiency or insulin resistance, diabetes mellitus, glucose intolerance, abnormal lipid metabolism, hyperglycemia, dyslipidemia, barrier function diseases including inflammatory bowel disease, Crohn's disease and ulcerative colitis, irritable bowel, disorder of intestinal contractility, hypercholesterolemia and atherogenic dyslipidemia, and in particular insulin resistance or obesity. Additionally, compositions of the invention are useful for promoting weight loss, reducing visceral fat, or improving gut barrier function. Additionally, the compositions of the invention are useful for controlling or normalizing blood glucose levels. Finally, the compositions of the invention are useful in the treatment of insulin resistance, prediabetes, type 1 diabetes or type 2 diabetes.

BACKGROUND Diabetes, obesity and overweight are a global trend, with the number of obese adults estimated at around 600 million. The prevalence of obesity-related disorders, such as, for example, diabetes, hypertension, heart disease and liver disease. Due to these highly debilitating pathologies, obesity is currently considered in Western countries as one of the most important public health problems. At the same time, the prevalence of type 2 diabetes and prediabetes is reaching pandemic proportions worldwide. Both the number of cases and the prevalence of diabetes have steadily increased over the past decades. About 422 million people worldwide have diabetes. Diabetes of all types can lead to complications, including kidney failure, leg amputation, loss of sight and nerve damage.

Obesity is associated with metabolic dysfunctions having an impact on glucose homeostasis and lipid metabolism, for example.

Another comorbidity of obesity and diabetes is a low-grade inflammatory condition associated with higher levels of blood lipopolysaccharides (LPS), also called metabolic endotoxemia.

Another comorbidity of obesity and diabetes is impaired or reduced gut barrier function.

Increased intestinal permeability leads to the translocation of bacteria and/or microorganism components into the bloodstream and into organs such as the liver or fatty tissue.

Intestinal inflammation, LPS and bacterial translocation are also seen in inflammatory bowel disease.

Examples include Crohn's disease, colitis, ulcerative colitis, bowel or colic pain, and other inflammatory bowel diseases.

Interestingly, both inflammatory bowel disease and obesity-related disease are associated with changes in the composition of the gut microbiota.

The human intestine is colonized by a diverse, complex and dynamic community of microbes representing more than 1000 different species, which constantly interact with the host.

The intestinal microbiota depends on the characteristics of the host such as age, sex, genetic background.

Other factors include environmental conditions such as stress, medications, gastrointestinal surgery, infectious and toxic agents, and daily dietary changes.

The gut microbiota impacts the development of diabetes, obesity and related disorders, overweight and gut inflammation, or gut pain.

Dysbiosis of the microbiota can further disrupt crosstalk between organs and gut barrier integrity, leading to symptoms.

Camu camu has also been suggested in the treatment of obesity, insulin resistance and steatosis.

In particular, camu camu would be associated with better health and greater bacterial diversity in the intestinal microbiota.

Camu camu is suggested to prevent visceral and hepatic fat deposition through activation of brown adipose tissue and increased energy expenditure. see: Anhê et al, Treatment with camu camu (Myrciaria dubia) prevents obesity by altering the gut microbiota and increasing energy expenditure in diet-induced obese mice, Gut, 68(3), 453 - March 2019, https://doi. org/10.1136/gutjnl-2017-315565. However, this document does not reveal a composition comprising Akkermansia muciniphila and camu camu extract.

WOO2013130773A2 to Kaplan discloses a method for altering the relative abundance of microbiota in a subject comprising administering to the subject an effective dose of a composition consisting essentially of substantially purified Verrucomicrobia for the treatment of obesity and metabolic syndrome.

WO2014076246 at the Catholic University of Louvain and Wageningen Universiteit describes the use of Akkermansia muciniphila for the treatment of obesity and related disorders. However, this document does not reveal a composition comprising Akkermansia muciniphila and camu camu extract.

EP3347030 at the Catholic University of Leuven and Wageningen Universiteit describes Akkermansia muciniphila for use in the treatment of a metabolic disorder in a subject in need thereof, wherein Akkermansia muciniphila is pasteurized. This document does not describe the combination of Akkermansia muciniphila with a plant extract.

However, there is still an urgent need to provide improved compositions for the treatment of metabolic disorders, in particular prediabetes, type 1 diabetes or type 2 diabetes.

BRIEF DESCRIPTION OF THE INVENTION The current inventors have surprisingly discovered that the combination of Akkermansia muciniphila and an extract of camu camu gives superior results in glucose control, disorders of insulin sensitivity or weight loss compared to other herbal extracts and is therefore synergistic. This is particularly the case when Akkermansia muciniphila is pasteurized.

Accordingly, a first aspect of the invention is a composition comprising Akkermansia muciniphila and a camu camu extract.

In another aspect of the invention, Akkermansia muciniphila is pasteurized.

Another aspect of the invention is a composition for use in the prevention or treatment of a metabolic disorder selected from the group comprising obesity, metabolic syndrome, disorders related to insulin deficiency or insulin resistance. to insulin, diabetes mellitus, glucose intolerance, abnormal lipid metabolism, hyperglycemia, dyslipidemia, barrier function diseases including inflammatory bowel disease, Crohn's disease and ulcerative colitis, irritable bowel syndrome, intestinal contractility disorder, hypercholesterolemia, intestinal barrier function disorder and atherogenic dyslipidemia. In another aspect of the invention, the metabolic disorder is prediabetes, type 1 diabetes or type 2 diabetes. In another aspect of the invention, the composition reduces the development of fat mass, in particular the visceral adipose tissue, compared to an untreated subject. In another aspect of the invention, the composition further comprises one or more ingredients selected from the group consisting of probiotics, bacteria, yeasts, microorganisms, prebiotics or a combination thereof. In another aspect of the invention, the composition further comprises a mineral or vitamin or a combination thereof. In another aspect of the invention, the composition further comprises B2, B3, B12 or a combination thereof. In another aspect of the invention, the composition further comprises a pharmaceutically acceptable carrier or a food grade carrier. In another aspect of the invention, the composition is administered orally. In another aspect of the invention, the pasteurized Akkermansia muciniphila is administered in an amount of 1.10% cells to 1.10"? cells per day, preferably 1.105 cells to 1.107" cells per day, and even more preferably from 1.106 to

1.10"° cells per day, preferably as expressed in TFU by flow cytometry. In another aspect of the invention, the camu camu extract is administered in an amount of 50 mg to 3000 mg per day, preferably , 100 mg to 2000 mg, even more preferably 200 mg to 1000 mg per day, and even more preferably 250 mg to 500 mg per day.

In another aspect of the invention, the composition is a cosmetic composition, a nutritional composition, a food product, a food supplement, a medical food or a drug.

In another aspect of the invention, the composition further comprises a second plant extract.

In another aspect of the invention, the second plant extract is chosen from the group consisting of Aronia melanocarpa, Emblica officinalis, Olea Europa, Citrus bergamia, Vaccinium macrocarpon, Myrciaria dubia, Panax ginseng rouge, Vaccinium oxycoccos, Vaccinium macrocarpon.

Another aspect of the invention is a method of producing the composition of the invention comprising the following steps a.

Cultures of cells of the genus Akkermansia; b.

Pasteurization of the cultured cells obtained in step a); vs.

Add cryoprotectant and pH buffer; d. lyophilization of the pasteurized cells from step c); e. mixing the freeze-dried powder with an extract of camu camu f.

Optionally, produce capsules, gel capsules or soft gel capsules from the mixture from step e). Another aspect of the invention is a composition according to one of the preceding claims for promoting weight loss or intestinal barrier function in a subject in need thereof.

Another aspect of the invention is a method of treating or preventing a metabolic disorder in a subject in need thereof, wherein the method comprises administering to the subject an amount effective, particularly without limiting side effects the treatment, of a composition comprising Akkermansia muciniphila and an extract of camu camu. In a preferred embodiment, Akkermansia muciniphila is pasteurized.

DEFINITIONS “Treatment” means the reduction or alleviation of one or more adverse effects or symptoms of a disease, disorder or condition. This term therefore designates both therapeutic treatment and prophylactic or preventive measures. “Prevention” means preventing a disease or condition from occurring or reducing the risk of it occurring. By "effective amount" or "therapeutically effective amount" is meant the level or amount of agent that is targeted, without causing significant negative or adverse side effects to the target, delaying or preventing the onset of a disorder metabolic, slowing or stopping the progression, aggravation or deterioration of one or more symptoms of the metabolic disorder; cause an improvement in the symptoms of the metabolic disorder; reduce the severity or incidence of the metabolic disorder; cure the metabolic disorder; or restore the normal quantity and/or proportion of Akkermansia muciniphila in the intestine of the subject to be treated. "Akkermansia muciniphila' refers to the mucin-degrading bacterium identified by Derrien (Derrien et al., 2004. Int. J. Syst. Evol. Microbiol. 54:1469-1476). The cells are oval-shaped, non-motile and Gram negative. Akkermansia muciniphila may also be referred to as Akkermansia spp. or Akkermansia-like bacteria. It belongs to the phylum Verrucomicrobia. It is generally accepted that strains with an experimentally determined nucleotide similarity determined by DNA-DNA hybridization of approximately 70% can be considered as the same species - corresponding to an average nucleotide identity (ANI) of approximately 95%.In one embodiment, Akkermansia muciniphila also includes fragments of Akkermansia muciniphila. "Pasteurized Akkermansia muciniphila" means Akkermansia muciniphila subjected to heat treatment In one version, pasteurized Akkermansia muciniphila means Akkermansia muciniphila that has been heated to a temperature in between 50°C and 100°C for at least 10 minutes.

"TFU" or "Total Fluorescent Units" means the number of cells as determined by their fluorescence brightness after staining. In a preferred embodiment, TFU is measured by flow cytometry. For example, in a first step, aliquots of batches of biomass are rehydrated in PBS and stained with Syto 9 and propidium iodide according to the manufacturer's protocol (LIVE/DEAD® BacLight TM Bacterial Viability Kit, Thermofisher). The TFU can then be obtained by analyzing the stained samples on the Attune NxT flow cytometer. In general, TFUs are expressed in total fluorescent units per gram or per ml, preferably per gram.

"Camu camu extract" means any product processed from any part of the plant, in particular the fruit of camu camu (Myrciaria dubia). Camus camu is an Amazonian fruit that is used as an antioxidant in foodstuffs. Camu camu extract can be obtained by drying, alcoholic extraction or water extraction, concentrated or not and transformed into liquid or powder. Water extraction is preferred. In another variant, the camu camu extract is a crude extract. In another variation, the camu camu extract has a vitamin C content of 10% or more by dry weight. In another variant, the camu-camu extract comprises polyphenols chosen from the group consisting of proanthocyanidins, flavanols, flavonoids and ellagitannins. The camu-camu extract may also include fiber.

“Probiotics” are live microorganisms which, when administered in effective amounts, have a beneficial effect on the health or well-being of a subject. In one instance, these health benefits are associated with improving the balance of human or animal microbiota in the gastrointestinal tract, or restoring a normal microbiota.

The term "prebiotic" refers to a substance, such as a substance which cannot be digested by humans, but which modulates the composition and/or activity of the intestinal microbiota by its metabolism by microorganisms in the intestine. , thereby conferring a beneficial physiological effect on the host.

The "subject" means an animal, preferably a mammal, more preferably a human or an animal.

The term "overweight" refers to a situation in which the subject has a body mass index (BMI) of 25 or more.

In this context, BMI is defined as an individual's body mass (in kg) divided by the square of their height (in meters). “Obesity” refers to a condition in which the subject has a BMI greater than or equal to 30. “Type | little or no insulin secretion.

"Prediabetes" means a condition where said subject has an elevated blood glucose level (giyceria) with a fasting glucose level greater than 100 mg/dl but less than 126 mg/dl or a 2-hour OGTT plasma glucose level greater than 140 mg/dl but less than 200 mg/dl.

"Type II diabetes" means a condition in which said subject has an elevated blood glucose (or blood sugar) level with a fasting blood glucose level greater than or equal to 126 mg/dl or a 2 hour blood glucose level on the glucose tolerance test orally (TGC) greater than or equal to 200 mg/dl. "Fragment" may refer to cellular components, metabolites, secreted molecules, vesicles and compounds resulting from the metabolism of Akkermansia muciniphila and others.

The fragments can be obtained, for example, by recovering supernatant from a culture of Akkermansia muciniphila or by extracting cell components or cell fractions, metabolites or secreted compounds from a culture of Akkermansia muciniphila.

DETAILED DESCRIPTION OF THE INVENTION The applicant demonstrates here the beneficial effects on the metabolism observed after administration of a composition comprising Akkermansia muciniphila and an extract of camu camu.

Accordingly, the present invention relates to a composition comprising Akkermansia muciniphila and a camu camu extract for the treatment, or for use in the treatment, of metabolic disorders in a subject in need thereof.

In the present document, a metabolic disorder is a disorder related to an alteration in metabolic homeostasis, such as, for example, an alteration in glucose or lipid homeostasis.

In one embodiment, said metabolic disorder is prediabetes, type 1 diabetes, or type 2 diabetes. insulin resistance, diabetes mellitus including type 1 diabetes or type 2 diabetes, glucose intolerance, abnormal lipid metabolism, atherosclerosis, hypertension, preeclampsia, heart disease, stroke, disease non-alcoholic fatty liver disease, hyperglycaemia, hepatic steatosis, dyslipidaemia, immune system dysfunction associated with overweight and obesity, liver diseases such as, for example, fibrosis associated with obesity, or liver function abnormalities including changes in bile production, inflammatory, immune and barrier function diseases such as, for example, inflammatory bowel disease, including disease Crohn's disease and ulcerative colitis, and irritable bowel syndrome, cardiovascular disease, hypercholesterolemia, elevated triglycerides, asthma, sleep apnea, osteoarthritis, neurodegeneration, gallbladder disease , syndrome X, inflammatory and immune disorders, atherogenic dyslipidemia and cancer.

In one case, hypercholesterolemia corresponds to a plasma cholesterol concentration greater than or equal to 2 g/L or 5 mmol/L.

In another variant, hypercholesterolemia corresponds to a ratio between the plasma concentration of total cholesterol and the plasma concentration of HDL (high density lipoprotein cholesterol) greater than or equal to 4.5:1, preferably 5:1. In one embodiment, the pasteurized Akkermansia muciniphila of the invention are non-viable cells.

In this document, “non-viable cells” refer to cells that are not able to proliferate.

Examples for visualizing or counting Akkermansia muciniphila cells were provided by Derrien et al. (2008. Appl.

About.

Microbiol. 74:1646-8), Derrien et al. (2011. Frontiers Microbiol. 2:166-175) or Reunanen et al. (2015. Appl.

About.

Microbiol. 81(11):3655-62).

The composition of the invention may also include a pharmaceutically acceptable excipient or food grade carrier, for example solvents, dispersing media, coatings, isotonic agents and absorption delaying agents, etc. For human administration, preparations must meet general standards of safety and purity, as required by FDA Office of Biologics standards. The present invention also relates to a drug, a medical food, a food or a dietary supplement comprising an effective amount of Akkermansia muciniphila or a fragment thereof and a camu camu extract.

The present invention also relates to a method for treating or preventing a metabolic disorder in a subject in need thereof, wherein said method comprises administering an effective amount of Akkermansia muciniphila or a fragment thereof and an excerpt from camu camu about who needs it.

Another object of the invention is a method for restoring a normal proportion of Akkermansia muciniphila, fragments or other active compounds of Akkermansia muciniphila in the intestine of a subject in need thereof, in which said method comprises administering an effective amount of Akkermansia muciniphila or a fragment thereof and a camu camu extract to the subject.

The composition of the invention is administered at least once a week, preferably at least twice a week, more preferably at least three times a week, and even more preferably at least four times a week. In another form, the composition of the invention is administered at least once a day, and preferably at least twice a day.

In one embodiment, the composition of the invention is administered for 1 week, preferably for 2, 3, 4, 5, 6, 7 or 8 weeks or more, or even permanently.

In one embodiment, the composition of the invention is administered for a period that lasts until the desired result is achieved, e.g. normal blood glucose levels, reduced insulin sensitivity, reduced prediabetes , reduced insulin resistance as expressed in HOMA-IR, reduced type 1 diabetes or reduced type 2 diabetes, weight loss, treatment of metabolic disorders or lower cholesterol level in the plasma. In one embodiment, pasteurized Akkermansia muciniphila is administered in an amount of 1.10* cells to 1.10"? cells per day, preferably 1.105 cells to 1.101! cells per day, and even more preferably 1.106 to 1.10 "° cells per day. In another version of the invention, the daily dose of pasteurized Akkermansia muciniphila is from 1.10° to around 1.10"? cells/day, preferably from around 1.108 to 1.10"° cells/day, and, even more, from preference of 1.10° at 1.107 cells/day.

The present invention also relates to the cosmetic use of pasteurized Akkermansia muciniphila or a fragment thereof to promote weight loss in a subject.

Another object of the invention is therefore a cosmetic composition comprising a cosmetically effective amount of pasteurized Akkermansia muciniphila or a fragment thereof and camu camu extract, and its use for promoting weight loss in a topic. By "cosmetically effective amount" is meant here the amount of a cosmetic composition necessary and sufficient to promote a cosmetic effect, such as for example to induce weight loss in a subject.

In one of the embodiments of the invention, the composition, the pharmaceutical composition, the cosmetic composition or the medicament further comprises additional probiotic strains or species, such as, for example, bacterial probiotic strains or species. In another form, the other strains are pasteurized. These probiotics include strains or species of bacteria or fungi, preferably strains or species of yeast. In a first variant, these additional probiotic strains or species are selected from those which are naturally present in the intestine of the subject, preferably in the human intestine, and more preferentially in the intestine of healthy human subjects.

Examples of bacterial probiotic strains or species that can be used in the present invention include, but are not limited to, Lactobacillus, Lactococcus, Bifidobacterium, Veillonella, Desemzia, Christensenella, Allobaculum, Coprococcus, Collinsella, Citrobacter, Turicibacter, Sutterella, Subdoligranulum, Streptococcus, Sporobacter, Sporacetigenium, Ruminococcus, Roseburia, Proteus, Propionobacterium, Leuconostoc, Weissella, Pediococcus, Streptococcus, Prevotella, Parabacteroides, Papillibacter, Oscillospira, Melissococcus, Dorea, Dialister, Clostridium, Cedecea, Catenibacterium, Butyrivibrio, Buttiauxella, Bulleidia, Bilophila, Bacteroides, Anaerovorax, Anaerostipes, Anaerofilum, _Enterobacteriaceae, Fermicutes, Atopobium, Alistipes, Acinetobacter, Slackie, Shigella, Shewanella, Serratia, Mahella, Lachnospira, Klebsiella, Idiomarina, Fusobacterium, Faecalibacterium, _Eubacterium, … Enterococcus, Enterobacter, Eggerosmobbella, Dygerosthella or Anaerobacterium.

Preferred probiotic strains are Lactobacillus, Lactococcus, Bifidobacterium, Christensenella, Clostridium, Anaerostipes, Faecalibacterium, Eubacterium, Enterococcus, Enterobacter, Eggerthella, Dysosmobacter or Anaerobacterium.

The strains or species of prokaryotes which can be used in the present invention include, among others, Archaea, Firmicutes, Verrucomicrobia, Christensenella, Bacteroidetes (such as, for example, Allistipes, Bacteroides ovatus, Bacteroides splachnicus, Bacteroides stercoris, Parabacteroides, Prevotella ruminicola, Porphyromondaceae, and related genera), Proteobacteria, Beta-proteobacteria (such as, for example, Aquabacterium and Burkholderia), Gamma-proteobacteria (such as, for example, Xanthomonadaceae), Actinobacteria (such as, for example, Actinomycetaceae and Atopobium), Fusobacteria, Methanobacteria , Spirochetes, Fibrobacteria, Deferribacteria, Deinococcus, Thermus, Cyanobacteria, Methanobrevibacteria, Peptostreptococcus, Ruminococcus, Coprococcus, Subdolingranulum, Dorea, Bulleidia, Anaerofustis, Gemella, Roseburia, Dialister, Anaerotruncus, Staphylococcus, Micrococcus, Propionobacteria, Enterobacteriaceae, Faecalibacterium, Bacteroides, Bacteroides , Prevotell a, Eubacterium, Bacilli (such as, for example, Lactobacillus salivarius and related species, Aerococcus, Granulicatella, Streptococcus bovis and related genus and Streptococcus intermedius and related genus), Clostridium (such as, for example, Anaerobutyricum hallii,

Eubacterium limosum, Anaerobutyricum soehngenii and related genus) and Butyrivibrio. Examples of fungal probiotic strains or species, preferably yeast probiotic strains or species, which may be used in the present invention include, but are not limited to, Ascomycetes, Zygomycetes and Deuteriomycetes, preferably Aspergillus groups, | Torulopsis, Zygosaccharomyces, Hansenula, Candida, Saccharomyces, Clavispora, Bretanomyces, Pichia, Amylomyces, Zygosaccharomyces, Endomycess, Hyphopicia, Zygosaccharomyces, Kluyveromyces, Mucor, Rhizopus, Yarrowia, Endomyces, Debaryomyces, and/or Penicillium.

In one embodiment, the only microbial strain or species, preferably bacterial, included in the composition, the pharmaceutical composition, the cosmetic composition or the drug is Akkermansia muciniphila.

In one embodiment, the composition, pharmaceutical composition, cosmetic composition or drug consists of pasteurized Akkermansia muciniphila.

In one embodiment of the invention, the composition, pharmaceutical composition, cosmetic composition or medicament further comprises a prebiotic.

Examples of prebiotics that can be used in the present invention include, but are not limited to, plant extracts containing polyphenols, linulin and inulin-like fructans, oligofructose, beta-glucans, xylose, arabinose, arabinoxylan, ribose, galactose, rhamnose, cellobiose, fructose, lactose, salicin, sucrose, glucose, aesculin, tween 80, trehalose, maltose, mannose, melibiose, mucus or mucins, raffinose, fructooligosaccharides, galacto-oligosaccharides, amino acids, alcohols, fermentable carbohydrates and any combinations thereof.

Other non-limiting examples of prebiotics include water-soluble cellulose derivatives, water-insoluble cellulose derivatives, unprocessed rolled oats, metamucil, bran, and any combination thereof.

Examples of water-soluble cellulose derivatives include, but are not limited to, methylcellulose, methylethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, cationic hydroxyethylcellulose, hydroxypropylcellulose, hydroxyethylmethylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose.

The composition of the invention can be administered orally, rectally, oesophagogastroduodenoscopy, colonoscopy, nasogastric or orogastric tube.

The composition can be administered orally in the form of tablets, pills, capsules, soft gelatin capsules, sugar coated pills, or as dispersing tablets, effervescent tablets or other solids.

The composition can be administered orally in liquid, drinkable solution or liposome form.

In one embodiment, the composition of the invention further comprises excipients, diluents and/or carriers chosen according to the intended route of administration.

Examples of excipients, diluents and/or carriers include, but are not limited to, water, phosphate buffered saline, anaerobic phosphate buffered saline, sodium bicarbonate, juice, milk , yoghurt, infant formula, dairy products, colorants, such as, for example, titanium dioxide (E171), iron dioxide (E172) and brilliant black BN (E151); flavoring agents; thickeners, such as for example glycerol monostearate; sweeteners; coating agents, such as for example refined rapeseed oil, soybean oil, peanut oil, soy lecithin or fish gelatin; diluents, such as, for example, lactose, lactose monohydrate or starch; binders, such as for example povidone, pregelatinized starch, gums, sucrose, polyethylene glycol (PEG) 4000 or PEG 6000; disintegrating agents, such as for example microcrystalline cellulose or sodium carboxymethyl starch, such as for example sodium carboxymethyl starch type A; lubricating agents, such as for example magnesium stearate; flow agent, such as anhydrous colloidal silica, etc.

The composition of the invention is a pharmaceutical composition.

In another case, the composition of the invention is a food additive, a beverage additive, a food supplement, a nutritional product, a medical food or a nutraceutical composition. Disorders of glucose homeostasis are associated with increased intestinal permeability and impaired mucus production, epithelial barrier, immune system and/or production of antibacterial compounds by the subject and inflammation of low intensity. Accordingly, the composition of the present invention can be used to decrease intestinal permeability, to restore impaired mucus production, to restore the epithelial barrier, to restore the immune system, or to restore the production of antibacterial compounds or a combination thereof. -this.

Additionally, the composition of the invention may be useful in controlling gut barrier function. Accordingly, another aspect is a method of controlling intestinal barrier function comprising administering an effective or cosmetically effective amount of the composition of the invention to a subject in need thereof. In one of these versions, the composition of the invention regulates the thickness of the mucus layer which can be reduced in case of obesity or other metabolic disorders. In another, the administration of the composition of the invention induces the production of colonic antimicrobial peptides, such as Reglllgamma, for example. In another case, the administration of the composition of the invention induces the production of compounds of the endocannabinoid family, such as for example the acylglycerols chosen from the group comprising 2-oleoylglycerol, 2-palmitoylglycerol and 2-arachidonoylglycerol . In another variation, administration of the composition of the invention regulates mucus turnover.

In one embodiment, the administration of the composition of the invention to a subject increases the satiety of said subject. Accordingly, according to this embodiment, the method of the invention increases satiety or decreases food intake in a subject, thereby inducing sustained weight loss in the subject, and thereby treating metabolic disorders in said subject, such as as, for example, obesity-related metabolic disorders.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Accelerated blood glucose at the end of treatment.

Effects of oral administration of vehicle or A. muciniphila pasteurized or with A. muciniphila and Aronia or A. muciniphila and Camu Camu in mice fasted for 6 hours on glycemia at the 11th week of treatment. **p<0.01 vs HFD vehicle (p-values were obtained using one-way ANOVA followed by Bonferroni's post-hoc test). Figure 2: Insulinemia at the end of treatment.

Effects of oral administration of vehicle or A. muciniphila pasteurized or A. muciniphila and Aronia or A. muciniphila and Camu Camu in mice fixed at 6h on plasma insulin level at week 11. **p<0.01 compared to vehicle HFD; Sp < 0.05 between 2 groups (*p values were obtained using one-way ANOVA followed by Bonferroni's post-hoc test; Sp values were obtained using t-test comparing 2 groups). Figure 3: HOMA index at the end of treatment.

Effects of oral administration of vehicle or A. muciniphila pasteurized or A. muciniphila and Aronia or A. muciniphila and Camu Camu in mice fixed at 6 h on the HOMA IR index calculated as insulinemia (mU/L) x blood sugar (mmol/L) / 22.5 at the start of the protocol at week 11 of treatment. **p<0.01 vs HFD vehicle; $$p < 0.01 between 2 groups (p-values were obtained using 1 or 2-way ANOVA followed by Bonferroni post-hoc test; Sp values were obtained using t-test comparing 2 groups ). Figure 4: Relative mRNA expression of gut barrier integrity biomarkers and antimicrobial peptides in the colon at the end of treatment.

The effects of oral administration of vehicle or pasteurized A. muciniphila or A. muciniphila and Aronia or A. muciniphila and Camu Camu at week 12 on various — markers of gut barrier integrity and antimicrobial peptides in the colon (RCPQ analysis of fold change versus mRNA analysis) are depicted in Figure 4. HFD Vehicle group) mRNA expression ZO-1 (A), Reg3y expression of mRNA (B), Intectin mRNA expression (C), Occludin mRNA expression (D), Claudin3 mRNA expression (E), Muc2 mRNA expression (F). *p < 0.05, *p < 0.01, ***p < 0.001 vs HFD Vehicle, #p < 0.05, ##p < 0.001 vs HFD pAkk, Sp between 2 groups (the values *p and #p were obtained using 1-way ANOVA followed by Bonferroni post-hoc, Sp values were obtained using t-test comparing 2 groups).

DETAILED DESCRIPTION OF THE DRAWING Figures 1, 2, 3 and 4 show the in vivo effect of pasteurized Akkermansia muciniphila (referred to as "pAkk" in the figures) alone or in combination with plant extracts on fasting blood glucose, fasting insulinemia, HOMA index and intestinal barrier function in mice fed a high fat diet, hereinafter referred to as "HFD". The nine-week-old male C57BL/6J mice (Charles River Laboratory, l'Arbresle, France) benefited from an acclimatization period of at least 5 days after their arrival. The animals were housed in ventilated and enriched cages (48 x 37.5 x 21 cm 3 ) throughout the experimental phase. Animal cage bedding was changed once a week. The mice were housed in groups of 5 animals according to a normal light cycle (at 7 p.m., lights off), 22 ± 2° C. and 50 ± 10% relative humidity. Hosting settings were recorded daily. During the acclimatization phase, the standard diet (RM1(E)801492, SDS) and tap water were provided ad libitum. During the experimental phase (12 weeks), a 45% high fat diet (Research Diet #12451) (HFD group) and tap water were provided ad libitum. Mice were treated daily with an oral gavage of 180 µl of vehicle, or 180 µl of A. muciniphila alone or 180 µl of solution of A. muciniphila + each plant extract (Aronia: 100 mg/kg; Camu Camu: 250 mg/kg) during the 12 weeks of HFD treatment. Before the start of the protocol, at weeks 0, 4, 8 and 11, blood glucose was assessed with a glucometer (Accu-Chek Active, Roche), and a blood sample was taken to perform an insulin test (method HTRF) after 6 hours of fasting. The homeostasis index was calculated as follows: blood insulin (mU/L) x blood sugar (mmol/L) / 22.5. At week 11, fasted mice were orally loaded with glucose (2 g/kg body weight), approximately 24 hours after the last oral administration of vehicle or treatments. Blood glucose was measured at -30.0 (time of oral glucose loading), and 15, 30, 60, 90, and 120 min with a glucometer (Accu-Chek Active, Roche) on blood samples taken at the end of the tail vein.

Blood was also collected from the tail vein at −30 and 15 min with heparinized capillaries (=60 μl) (sampling of two capillaries per mouse). The blood was centrifuged at 13000g for 5 min at 4°C.

At least 15 µL of plasma was stored in a 0.5 ml tube at -80°C until further analyzes were performed.

The proximal colon was individually homogenized and tissue total RNA was prepared using TRIReagent (Sigma Aldrich) and QlAcube RNeasy 96 HT Kit (Qiagen). Total RNA quantification analysis was performed by analyzing 1 µl of each sample in Nanodrop (Ozyme). cDNA was prepared by reverse transcription of 1 µg of total RNA using an iScript RT kit (Biorad). Real-time PCR was performed with the Via7 real-time PCR system and software (applied biotechnology) using SYBR Green real-time PCR master mixes (Biorad) for detection.

The RPL19 gene was chosen as a maintenance gene.

All samples were run in duplicate, and data were analyzed using the 2AACT method.

The identity and purity of the amplified product was assessed by analysis of the melting curve at the end of the amplification.

It is surprising to note that among the plant extracts tested, Camu Camu is the one which, combined with pasteurized Akkermansia muciniphila, gave superior results on fasting glycaemia, insulinaemia and the HOMA index (figures 1 , 2 and 3), as well as on intestinal barrier function (Figure 4). The combination of pasteurized Akkermansia muciniphila and an extract of Camu Camu also improved the barrier function with a more pronounced effect on the expression of Reg-3-, claudin-3 and Muc2 mRNA (see figure 4) . These results indicate a potentiating or synergistic effect of Camu Camu extract when combined with pasteurized Akkermansia muciniphila.

Claims (15)

1. Composition comprising Akkermansia muciniphila and a camu camu extract. 2. Composition according to claim 1, in which Akkermansia muciniphila is pasteurized. 3. Composition according to one of the preceding claims, intended for use in the prevention or treatment of a metabolic disorder chosen from the group comprising obesity, metabolic syndrome, disorders linked to insulin deficiency or insulin resistance, diabetes mellitus, glucose intolerance, abnormal lipid metabolism, hyperglycemia, dyslipidemia, barrier function diseases including inflammatory bowel disease, Crohn's disease and ulcerative colitis , irritable bowel syndrome, intestinal contractility disorder, hypercholesterolemia and atherogenic dyslipidemia. 4. Composition according to one of the preceding claims, in which the metabolic disorder is prediabetes, type 1 diabetes or type 2 diabetes. 5. Composition according to one of the preceding claims, further comprising one or more ingredients chosen from the group consisting of probiotics, bacteria, yeasts, microorganisms, prebiotics or a combination thereof. 6. Composition according to one of the preceding claims, in which the composition further comprises a mineral or a vitamin or a combination thereof. 7. Composition according to one of the preceding claims, in which the mineral is chromium. 8. Composition according to one of the preceding claims, in which the vitamin is chosen from the group consisting of B2, B3, B12, or a combination thereof. 9. Composition according to one of the preceding claims, in which the composition further comprises a pharmaceutically acceptable carrier or a food-grade carrier. 10.Composition according to one of the preceding claims, in which the composition is administered orally. 11.Composition according to one of the preceding claims, in which the pasteurized Akkermansia muciniphila is administered in an amount of 1.10° cells to 1.10 cells per day, preferably from 1.10° cells to 1.101! cells per day, and more preferably from 1.106 to 1.101° cells per day. 12. Composition according to one of the preceding claims, in which the camu camu extract is administered in an amount of 50 mg to 3000 mg per day, preferably 100 mg to 500 mg, even more preferably 200 mg to 300 mg per day. 13.Composition according to one of the preceding claims, in which the composition is a cosmetic composition, a nutritional composition, a food product, a food supplement, a medical food or a drug. 14. Method for producing a composition according to one of the preceding claims comprising the following steps a. Cultures of cells of the genus Akkermansia; b. Pasteurization of the cultured cells obtained in step a); vs. Addition of cryoprotectant and pH buffer; d. lyophilization of the pasteurized cells from step c); e. mixing the freeze-dried powder with an extract of camu camu f. Optionally, produce capsules or gelatin capsules from the mixture from step e). 15.Use of the composition according to one of the preceding claims for controlling the blood glucose level in a subject who needs it.