Composition for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota

Abstract

The invention relates to the use of compositions comprising several specific molecules to treat pathological dysbiosis of the gut microbiota. Pathological dysbiosis of the gut microbiota can be characterized by an excess of sulfide-producing bacteria and a deficiency of butyrate-producing bacteria.

Description

Title of the invention: Composition for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota technical field

[0001] The invention relates to the prevention and / or treatment of pathological dysbiosis of the gut microbiota. In particular, the invention relates to the use of compositions comprising several specific molecules to treat pathological dysbiosis of the gut microbiota. Previous art

[0002] The microbiota is the collection of microorganisms (bacteria, archaea; viruses and eukaryotes) that develop in a specific environment.

[0003] The human body contains a multitude of microbiomes, particularly in the skin, mouth, vaginal cavity, throat and digestive system.

[0004] The microbiota present in the digestive system, also called intestinal microbiota, is distributed from the stomach to the colon and represents between 35 and 50% of the volume of intestinal contents for a weight greater than 1 kilogram.

[0005] The intestinal microbiota is the microbiota that houses a large part of the 10 trillion microorganisms populating the human body.

[0006] Although discovered in the 19th century, it was only with the rise of high-throughput sequencing in the 2000s that scientists were able to identify all the genes of the gut microbiota. The characterization of all the microbial genomes found in the gut led to the discovery of more than a thousand different species, the vast majority of which are bacterial.

[0007] In-depth analysis of the gut microbiota has demonstrated that, like a fingerprint, the gut microbiota is unique to each individual and depends on their environment as well as their genetic heritage.

[0008] Once established, the composition of the microbiota remains relatively stable throughout an individual's life. Variations are generally less than 5% compared to the overall microbiota.

[0009] Thus, pathogenic states of the intestinal microbiota are difficult to detect because they originate from these particular species.

[0010] When a patient has a disease, an imbalance of the pathogenic intestinal flora called pathological dysbiosis may be found; the threshold of imbalance is difficult to fix, as are the variations of genera and species unless there is a biomarker of the disease.

[0011] Thanks to the removal of technical barriers, the analysis of the intestinal microbiota has been able to reveal its major role both in maintaining the good health of the body and in the onset of many diseases.

[0012] Indeed, the gut microbiota plays an essential role in the functioning of the body. The microorganisms that compose it have a wide range of action, from the breakdown of food to epithelial repair and the regulation of metabolism.

[0013] Nevertheless, through dysbiosis, that is to say a change in the composition of the microbiota, the microbiota can be associated with derived or induced pathological disorders such as neurodegenerative diseases, intestinal diseases and certain cancers.

[0014] The interaction between the central nervous system and the gut is a major component in the development of certain pathologies. In Parkinson's disease, a link has been demonstrated between pathological dysbiosis of the gut microbiota and the onset of motor disorders as well as neuroinflammation.

[0015] Pathological dysbiosis of the intestinal microbiota has been demonstrated in other neurodegenerative diseases, particularly in patients with amyotrophic lateral sclerosis.

[0016] Furthermore, it is now known that intestinal diseases and more particularly inflammatory bowel diseases, Crohn's disease and food intolerance, especially gluten intolerance, are linked to an intestinal microbiota that is very depleted in microorganisms and therefore the presence of pathological dysbiosis.

[0017] Currently, the solutions proposed to prevent or combat dysbiosis consist mainly of administering probiotics and / or prebiotics, particularly to treat intestinal diseases, but their effectiveness is highly variable and depends on the patient's basic intestinal flora.

[0018] Another promising solution for combating pathological dysbiosis is fecal microbiota transplantation. However, the effect on the gut microbiota requires at least two transplants per week to achieve a lasting effect.

[0019] Furthermore, with regard to degenerative diseases specifically, these are often very debilitating for the patient. Some symptoms can be alleviated, but these diseases are never completely cured, and there is currently no satisfactory treatment. Many chemical agents have therapeutic properties, but their toxicity, half-life, or rapid elimination make them unsuitable for treating degenerative diseases.

[0020] Indeed, while these diseases are multifactorial, current treatments target only one of the factors causing them and do not seek to act more holistically with a systemic approach that directly targets the causes of the disease. For example, Charcot's disease, or amyotrophic lateral sclerosis, is linked to Numerous factors, particularly oxidative stress, mitochondrial dysfunction, neuroinflammation, excitotoxicity, oligodendrocyte dysfunction and degeneration, impaired proteostasis, an impaired DNA repair system, alterations in nucleocytoplasmic RNA and RNA bound to transport proteins, defective axonal transport, defective vesicular transport, and pathological dysbiosis of the gut microbiota, are involved. However, currently, the recommended treatment for ALS (Lou Gehrig's disease) is Riluzole®, which acts solely on inhibiting glutamate release to combat excitotoxicity, and no action on the gut microbiota is considered. The same observation applies to other neurodegenerative diseases.

[0021] There is therefore an important need for a solution to prevent or treat pathological dysbiosis of the intestinal microbiota, in particular to treat or prevent neurodegenerative or intestinal diseases, to meet an essential demand for new therapies.

[0022] The objective of the invention is therefore to meet all of these needs and to overcome the disadvantages and limitations of the prior art. Summary of the invention

[0023] To meet this objective, the invention proposes the use of particular compositions for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota, and adapted for uses to treat or prevent diseases associated with pathological dysbiosis of the intestinal microbiota, in particular neurodegenerative or intestinal diseases.

[0024] To this end, the invention relates to a composition comprising at least: - lactic acid, and / or a salt and / or an ester and / or anhydride of lactic acid, - butyric acid, and / or a salt and / or an ester and / or anhydride of butyric acid, and - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid,

[0025] for its use in humans or animals in the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0026] Advantageously, the composition according to the invention is capable of restoring the balance of the microbiota of an individual with pathological dysbiosis of the intestinal microbiota.

[0027] Preferably, the composition according to the invention is used to treat a pathological dysbiosis of the intestinal microbiota characterized in the intestinal microbiota by an excess of sulfur-producing bacteria and a deficiency of butyrate-producing bacteria.

[0028] Said treatment according to the invention is thus preferentially characterized in the intestinal microbiota by an increase in the proportion of butyrate-producing bacteria and a decrease in the proportion of sulfide-producing bacteria.

[0029] Advantageously, the effect on butyrate-producing bacteria and sulfide-producing bacteria provides a synergistic action to treat pathological dysbiosis and in particular pathological dysbioses associated with pathologies.

[0030] More specifically, the composition can be used in humans or animals for the prevention and / or treatment of at least one neurodegenerative disease and / or an intestinal disease associated with pathological dysbiosis of the gut microbiota. According to a particularly suitable variant, the composition can be used in humans or animals for the prevention and / or treatment of at least one neurodegenerative disease and / or an intestinal disease associated with pathological dysbiosis of the gut microbiota, in which the human or animal presents with intestinal dysbiosis characterized in the gut microbiota by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria.

[0031] Surprisingly, the inventors discovered that the composition according to the invention made it possible to treat or prevent diseases associated with pathological dysbiosis, and in particular neurodegenerative diseases or intestinal diseases associated with pathological dysbiosis such as lateral sclerosis or intestinal diseases such as Crohn's disease.

[0032] The composition according to the invention can thus be used as a drug or food supplement in humans or animals.

[0033] According to one variant, the composition according to the invention can be used to prevent or treat a neurodegenerative disease associated with a pathological dysbiosis of the intestinal microbiota chosen from amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease.

[0034] According to another variant, the composition according to the invention can be used to prevent or treat an intestinal disease associated with a pathological dysbiosis of the intestinal microbiota chosen from Crohn's disease, chronic inflammatory bowel diseases, ulcerative colitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis and food intolerance in particular gluten intolerance.

[0035] According to a particularly suitable embodiment, the composition for its use according to the invention, in addition to lactic acid, butyric acid and propionic acid, and / or a salt and / or an ester and / or an anhydride of these molecules, may also comprise at least one molecule selected from: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - famesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - acetic acid, and their combination the said molecule(s) may be in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0036] Preferably, the composition for its use comprises the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - famesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - acetic acid, - butyric acid, -lactic acid, - propionic acid and / or a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0037] Advantageously, each of these molecules acts on different factors, allowing for a multifactorial action on the gut microbiota. These molecules will thus help to better balance bacterial populations that are in excess or deficient, particularly by reducing intestinal inflammation.

[0038] Advantageously, the molecules of the composition for its use according to the invention can be conjugated to specific polymers, such as poly- lysine, thus increasing the efficacy and bioavailability of these molecules.

[0039] Thus, according to a particularly preferred embodiment, the composition for its use according to the invention comprises at least the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine: - one or more Butyrate-Poly-L-Lysine conjugates - one or more Lactate-Poly-L-Lysine conjugates - one or more Propionate-Poly-L-Lysine conjugates.

[0040] According to one variant, the composition for its use according to the invention comprises at least: - A. the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine: - one or more Oleyl-Poly-L-Lysine conjugates - one or more Palmitic-Poly-L-Lysine conjugates - one or more Lauryl-Poly-L-Lysine conjugates - one or more Azelayl-Poly-L-Lysine conjugates - one or more Palmitoleyl-Poly-L-Lysine conjugates - one or more Thioctyl-Poly-L-Lysine conjugates - one or more Myristyl-Poly-L-Lysine conjugates - one or more Orotyl-Poly-L-Lysine conjugates - one or more Acetate-Poly-L-Lysine conjugates - one or more Butyrate-Poly-L-Lysine conjugates - one or more Lactate-Poly-L-Lysine conjugates - one or more Propionate-Poly-L-Lysine conjugates - one or more Linoleyl-Poly-L-Lysine conjugates, and - B. of famesyl cysteine ​​and cholesterol, and / or an ester of these molecules, encapsulated in micelles, preferably in micelles formed by one or more of the conjugates listed in list A.

[0041] The invention also relates to a method for manufacturing these compositions.

[0042] Other features and advantages will become apparent from the detailed description of the invention and the examples that will follow. Brief description of the figures

[0043] [Fig-1] is a graphical representation of a PLS-DA analysis comparing the microbiota of WT and SOD1 mice

[0044] [Fig.2] is a graphical representation of a comparative analysis of the beta diversity of the intestinal microbiota of mice via the MDS-Jaccard method as a function of treatment.

[0045] [Fig.3] is a graphical representation of a comparative analysis of the beta diversity of mouse intestinal microbiota via the MDS - Bray Curtis method as a function of treatment.

[0046] [Fig.4] is a graphical representation of a comparative analysis of the microbiota in mouse testinal status as a function of treatment via PLS-DA analysis.

[0047] [Fig.5] is a graphical representation of a correlation analysis between mice WT or SOD1 depending on the bacterial families.

[0048] [Fig.6] is a graphical representation of a correlation analysis between mice SOD1 treated or untreated at low dose Detailed description of the invention

[0049] Definitions

[0050] For the purposes of this invention, "Animal" means any animal except for human beings (Man). This includes, in particular, mammals.

[0051] By "amphiphilic conjugate" in the sense of the invention is meant a conjugate formed by a hydrophobic molecule X and a hydrophilic polymer Y, the conjugate thus exhibiting an amphiphilic characteristic.

[0052] By "molecule X conjugated to a polymer Y" in the meaning of the invention, we mean a molecule X linked by a covalent bond to a polymer Y, said bond being preferentially an amide, urea or carbamate bond depending on the chemical nature of molecule X and polymer Y.

[0053] For the purposes of this invention, "medical nutrition composition," "medical nutrition product," "medical food," "food for special medical purposes" (FSMP), or "dietary food for special medical purposes" (DFSP) refers to a food intended for therapeutic prevention and / or treatment, used alone or in combination with other therapies. It is a nutritional compound, designed to address a specific clinical situation, that can constitute the sole or partial diet of the patients for whom it is intended.

[0054] By "pathological dysbiosis" in the sense of the invention is meant a pathological state characterized by an imbalance in the distribution of bacteria of the intestinal microbiota at the level of phyla, classes, orders, families, genera or species, leading to a risk of other diseases whose origin is metabolic.

[0055] For the purposes of this invention, "medicine" means a product which has been granted marketing authorization as a product for the prevention and / or treatment of a disease.

[0056] For the purposes of this invention, "microbiota" means the intestinal microbiota.

[0057] For the purposes of this invention, "butyrate-producing bacteria" means bacteria capable of producing butyrate through the anaerobic fermentation of indigestible dietary fiber. Butyrate-producing bacteria belong primarily to the following group: tarity to the phylum of Firmicutes.

[0058] For the purposes of this invention, "sulfide-producing bacteria" means bacteria capable of reducing sulfates into sulfides.

[0059] By "alpha diversity" in the sense of the invention, we mean the measure of the number of species present in a given sample.

[0060] By "beta diversity" in the sense of the invention, we mean the measure of the diversity of species in a given sample.

[0061] By "excess of sulfide-producing bacteria" in the sense of the invention, we mean a proportion of sulfide-producing bacteria greater than normal, preferably greater than the proportion of sulfide-producing bacteria in an individual not exhibiting pathological dysbiosis.

[0062] By "butyrate-producing bacteria deficiency" in the sense of the invention, we mean a proportion of butyrate-producing bacteria lower than normal, preferably lower than the proportion of butyrate-producing bacteria in an individual not exhibiting pathological dysbiosis.

[0063] Composition for its use

[0064] The invention therefore relates to a composition comprising at least: - lactic acid, and / or a salt and / or an ester and / or anhydride of lactic acid, - butyric acid, and / or a salt and / or an ester and / or anhydride of butyric acid, and - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid, for its use in humans or animals for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0065] Preferably, the composition is particularly suitable for use in humans for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0066] Advantageously, the combination of these three molecules ensures a significant preventive and / or curative effect on pathological dysbiosis of the gut microbiota. Lactate, butyrate, and propionate are the main short-chain fatty acids produced from the breakdown of fiber by gut microbiota bacteria. These three fatty acids are involved in numerous cellular functions and are the main energy source for the intestinal epithelium, particularly the epithelial cells of the intestine and colon. Insufficient levels of short-chain fatty acids can induce pathological dysbiosis.

[0067] In the context of the invention, pathological dysbiosis of the intestinal microbiota is preferentially characterized by an excess of sulfide-producing bacteria and a deficiency in butyrate-producing bacteria.

[0068] The presence of an excess of sulfur-producing bacteria is characteristic of pathological dysbiosis. Indeed, a decrease in the proportion of beneficial microorganisms within the microbiota leads to the emergence of bacterial populations that have a harmful effect or produce metabolites that are detrimental to the body. Thus, patients with pathological dysbiosis exhibit high levels of sulfur-reducing bacteria associated with an increased production of toxic substances such as hydrogen sulfide, known for its pro-inflammatory activity. The presence of these metabolites causes inflammation and increased intestinal permeability.

[0069] The use of a composition according to the invention is thus characterized in the intestinal microbiota by an increase in the proportion of butyrate-producing bacteria and a decrease in the proportion of sulfide-producing bacteria.

[0070] Advantageously, reducing the proportion of sulfur-producing bacteria helps to reduce intestinal inflammation by treating pathological dysbiosis. On the other hand, increasing the proportion of butyrate-producing bacteria allows the epithelial cells of the intestine and colon to restore membrane permeability. This increase stimulates the microbiota, particularly beneficial bacterial species. Thus, the simultaneous action on butyrate- and sulfur-producing bacteria results in a synergistic effect on pathological dysbiosis of the intestinal microbiota. The composition ensures a long-term effect on the intestinal microbiota by targeting the causes of pathological dysbiosis.

[0071] The composition can be used to prevent or treat any diseases that have caused or been caused by pathological dysbiosis of the intestinal microbiota, preferably pathological dysbiosis of the intestinal microbiota characterized in the microbiota by an increase in the proportion of sulfur-producing bacteria and a decrease in the proportion of butyrate-producing bacteria, and also preferably characterized by an increase in intestinal inflammation.

[0072] More specifically, the composition comprising at least: - lactic acid, and / or a salt and / or an ester and / or anhydride of lactic acid, - butyric acid, and / or a salt and / or an ester and / or anhydride of butyric acid, and - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid can be used in humans or animals for the prevention and / or treatment of at least one disease associated with pathological dysbiosis of the gut microbiota, of at least one disease associated with pathological dysbiosis of the gut microbiota in particular in which the human or animal presents intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfur-producing bacteria and a decrease in the proportion of butyrate-producing bacteria, and in particular at least one neurodegenerative disease and / or intestinal disease associated with pathological dysbiosis of the intestinal microbiota, such as at least one neurodegenerative disease and / or intestinal disease associated with pathological dysbiosis of the intestinal microbiota in which the human or animal presents intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfur-producing bacteria and a decrease in the proportion of butyrate-producing bacteria.

[0073] In the context of the invention, neurodegenerative diseases can be chosen from multiple sclerosis, amyotrophic lateral sclerosis, Parkinson's disease and Alzheimer's disease.

[0074] Furthermore, intestinal diseases can be chosen from Crohn's disease, chronic inflammatory bowel diseases, ulcerative colitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis and food intolerance, including gluten intolerance.

[0075] According to one variant, the composition can be used to treat or prevent autism spectrum disorders or spondyloarthritis, in particular in which the human or animal has intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfur-producing bacteria and a decrease in the proportion of butyrate-producing bacteria.

[0076] The composition thus treats pathological dysbiosis in particular by relatively increasing the proportion of butyrate-producing bacteria in the intestinal microbiota combined with a decrease in the proportion of sulfide-producing bacteria.

[0077] The composition thus acts on the intestinal epithelium by providing short-chain fatty acids essential for the functioning of epithelial cells, such as butyric acid. The epithelial cells will act on inflammation and treat pathological dysbiosis by promoting the growth of butyrate-producing bacteria while restoring membrane permeability.

[0078] The composition can thus be used as a drug, medical nutrition product or food supplement to rebalance the intestinal microbiota by increasing the proportion of sulfur-producing bacteria and decreasing the proportion of butyrate-producing bacteria and / or to prevent and combat diseases arising from or caused by this pathological imbalance of the intestinal microbiota.

[0079] In the context of the invention, sulfide-producing bacteria can be bacteria belonging to the Desulfovibrionaceae family and / or proteolytic bacteria.

[0080] Sulfur-producing bacteria belonging to the Desulfovibrionaceae family may belong to at least one bacterial genus selected from: - Alteridesulfovibrio - Aminidesulfovibrio - Bilophila - Desulfocurvus - Desulfovibrio - Frigididesulfovibrio - Fundidesulfovibrio - Halodesulfovibrio - Humidesulfovibrio - Lawsonia - Maridesulfovibrio - Megalodesulfovibrio - Nitratidesulfovibrio - Oleidesulfovibrio - Paradesulfovibrio - Paucidesulfovibrio - Pseudodesulfovibrio - Solidesulfovibrio.

[0081] Preferably, the sulfide-producing bacteria belong to the family Desulfovibrionaceae, genus Bilophila, and species wadworthia.

[0082] Thus, the composition according to the invention can be used in humans or animals to decrease the proportion of the bacterial species Bilophila wadworthia in humans or animals exhibiting intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfide-producing bacteria.

[0083] The composition also acts on at least one family of butyrate-producing bacteria selected from: - The Lachnospiraeceae - The Ruminococcacea

[0084] Butyrate-producing bacteria belonging to the Lachnospiraeceae family may belong to at least one bacterial genus selected from: - Lachnospira - Roseburria - Eubacterium - Anaerostipes.

[0085] Butyrate-producing bacteria belonging to the Ruminococcacea family may belong to the bacterial genus Faecalibacterium.

[0086] In a particularly preferred embodiment, the composition can be used to increase the proportion of bacteria from the Lachnospiraeceae family and decrease the proportion of bacteria from the Desulfovibriaceae family preferentially so that the Desulfovibriaceae represent less than 0.01% of the total bacteria present in the intestinal microbiota.

[0087] Advantageously, the composition makes it possible to reduce inflammation of the digestive system by restoring sulfide and butyrate concentrations similar to those of a healthy patient.

[0088] Preferably, the composition, in addition to lactic acid, butyric acid and propionic acid, and / or a salt and / or an ester and / or an anhydride of these molecules, also comprises at least one molecule selected from: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - Farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid - acetic acid, and their combination, the said molecule(s) may be in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0089] Preferably the composition comprises at least 2 molecules, in particular at least 3, even more preferably at least 4 chosen from: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - Farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid - acetic acid, and their combination, the said molecule(s) being in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0090] According to a particular embodiment of the invention, the composition, in addition to lactic acid, butyric acid and propionic acid, and / or a salt and / or an ester and / or an anhydride of these molecules, also comprises at least azelaic acid, cholesterol, myristic acid as well as lauric acid and / or a salt and / or an ester and / or an anhydride of these molecules.

[0091] For all molecules present in the compositions for its use according to the invention, when they are mentioned in this application, it may be the molecules (example butyric acid) as such and / or a salt (example: butyrate) and / or an ester and / or an anhydride of these molecules (example: butyric acid ester) and / or an anhydride.

[0092] According to a particularly preferred embodiment of the invention, the composition comprises at least the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - Farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - lactic acid - propionic acid -butyric acid - acetic acid, and their combination, the said molecule(s) may be in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0093] Such a composition, surprisingly and unexpectedly, acts on the intestinal microbiota and in particular is capable of increasing the proportion of bacteria producing butyrate and decrease the proportion of sulfide-producing bacteria.

[0094] The useful composition according to the invention comprising at least all of the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - Farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - lactic acid - propionic acid -butyric acid - acetic acid, and their combination, the said molecule(s) may be in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules, is referred to below as composition Cl.

[0095] The useful composition according to the invention, and in particular the Cl composition, may optionally also include pyruvic acid and / or a salt and / or an ester and / or an anhydride.

[0096] Preferably: - the amount of butyric acid in the composition, and more specifically in the Cl composition, is greater than that of each of the other molecules taken individually, and / or - the amount of thioctic acid in the composition, and more specifically in the Cl composition, is greater than that of each of the other molecules taken individually, except for butyric acid, and / or - the amount of lauric acid in the composition, and more particularly in the Cl composition, is greater than that of each of the following molecules taken individually: palmitic acid, linoleic acid, azelaic acid, farnesyl cysteine, palmitoleic acid, cholesterol, myristic acid, orotic acid, - the quantity of oleic acid on the one hand and lactic acid on the other hand in the composition, and more particularly in the Cl composition, is greater than that of each of the following molecules taken individually: palmitic acid, lactic acid lauric acid, linoleic acid, azelaic acid, farnesyl cysteine, palmitoleic acid, cholesterol, myristic acid, orotic acid, acetic acid.

[0097] Preferably, each of the molecules in the Cl composition represents between 0.5 E-05 M and 10 E-05 M.

[0098] According to a particular embodiment of the invention, the useful composition according to the invention, and in particular the Cl composition, can be used by administering a dose of between 0.5 and 10 mg / mL, in particular between 2 and 8 mg / mL, preferably 7 mg / mL to treat pathological dysbiosis of the intestinal microbiota.

[0099] According to one variant, the useful composition according to the invention, and in particular the Cl composition, is administered with a first so-called "attack" dose of between 7 and 10mg / mL, preferably between 9 and 10mg / mL, then a "maintenance" dose of preferably between 0.5 and 5mg / mL, in particular between 2 and 3mg / mL.

[0100] Preferably, the useful composition according to the invention, and in particular Cl, may comprise at least one polymer selected from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0101] More particularly, to improve the solubility, efficiency and bioavailability of the molecules of the useful composition according to the invention, at least one of the molecules of the composition chosen from butyric acid, lactic acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer chosen from poly-lysine, polyethylene glycol, polyomithine, polyarginine and polyhistidine.

[0102] When the composition is composition Cl or a composition comprising the molecules of the list below, preferably at least one of the molecules of the composition chosen from oleic acid, palmitic acid, lauric acid, linoleic acid, azelaic acid, palmitoleic acid, thioctic acid, myristic acid, orotic acid, acetic acid, butyric acid, lactic acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer chosen from poly-lysine, polyethylene glycol, polyomithine, polyarginine and polyhistidine.

[0103] Indeed, the molecules chosen from oleic acid, palmitic acid, lauric acid, linoleic acid, azelaic acid, palmitoleic acid, thioctic acid, myristic acid, orotic acid, acetic acid, butyric acid, lactic acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, are all covalently conjugatable with a polymer chosen from poly-lysine, polyethylene glycol, polyomithine, polyarginine and polyhistidine, in particular by an amide, urea or carbamate bond.

[0104] According to a particularly suitable variant, all molecules selected from oleic acid, palmitic acid, lauric acid, linoleic acid, azelaic acid, palmitoleic acid, thioctic acid, myristic acid, orotic acid, acetic acid, butyric acid, lactic acid, propionic acid, salts of these acids and esters of these acids, when present in the useful composition according to the invention, are covalently conjugated to at least one molecule of a polymer selected from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0105] The useful composition according to the invention may include micelles. When the composition according to the invention, and in particular composition Cl, includes famesyl cysteine ​​and / or cholesterol and / or one of their esters and / or salts and / or anhydrides, the useful composition according to the invention preferably includes micelles in which at least famesyl cysteine ​​and / or cholesterol and / or one of their esters and / or salts and / or anhydrides are encapsulated.

[0106] According to one embodiment, the useful composition according to the invention preferably comprises: - conjugates of oleic acid, palmitic acid, lauric acid, linoleic acid, azelaic acid, palmitoleic acid, thioctic acid, myristic acid, orotic acid, acetic acid, butyric acid, lactic acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, with poly-lysine, polyethylene glycol, polyomithine, polyarginine or polyhistidine, and - micelles formed by at least one of these conjugates, said micelles encapsulating famesyl cysteine ​​and cholesterol.

[0107] Preferably, at least one of the micelles is formed by amphiphilic conjugates, each consisting of at least one hydrophobic molecule covalently conjugated to a molecule of a polymer selected from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0108] Thus, according to a particular embodiment of the invention, at least one micelle is formed by amphiphilic conjugates, each consisting of at least one molecule chosen from oleic acid, palmitic acid, lauric acid, linoleic acid, palmitoleic acid, myristic acid, salts, esters and anhydrides of these fatty acids, covalently conjugated to a molecule of a polymer chosen from poly-lysine, polyethylene glycol, polyomithine, polyarginine and polyhistidine.

[0109] This configuration allows in particular: - to increase the half-life of the active molecules in the composition within the body, - to target the tissues or cells on which the composition molecules must act - to increase the stability and bioavailability of the active ingredient.

[0110] The effectiveness of the composition is therefore greater and it is possible to administer lower doses and reduce the acute or chronic toxicity of the active molecules contained in the composition.

[0111] Preferably, the polymer(s) conjugated to the molecules are chosen from poly-L-lysine, poly-ethylene-glycol, poly-L-ornithine, poly-L-arginine and poly-L-histidine.

[0112] The poly-lysine used in the compositions according to the invention is preferably a poly-L-lysine. The poly-lysine is preferably linear. In particular, the poly-lysine used may be an epsilon poly-L-lysine, preferably a poly-L-lysine with a molecular weight between 12,000 and 20,000 Da. The poly-lysine used in the compositions according to the invention may be a poly-lysine containing a bromide, chlorine, or trifluoroacetic acid counterion.

[0113] Thus, according to one variant, composition Cl comprises at least: -A of the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine: - one or more Oleyl-Poly-L-Lysine conjugates - one or more Palmitic-Poly-L-Lysine conjugates - one or more Lauryl-Poly-L-Lysine conjugates - one or more Azelayl-Poly-L-Lysine conjugates - one or more Palmitoleyl-Poly-L-Lysine conjugates - one or more Thioctyl-Poly-L-Lysine conjugates - one or more Myristyl-Poly-L-Lysine conjugates - one or more Orotyl-Poly-L-Lysine conjugates - one or more Acetate-Poly-L-Lysine conjugates - one or more Butyrate-Poly-L-Lysine conjugates - one or more Lactate-Poly-L-Lysine conjugates - one or more Propionate-Poly-L-Lysine conjugates - one or more Linoleyl-Poly-L-Lysine conjugates, - B: farnesyl cysteine ​​and cholesterol, and / or an ester of these molecules, encapsulated in micelles, preferably in micelles formed by one or more of the conjugates listed in list A.

[0114] In this composition, poly-L-Lysine can be replaced by another poly-lysine or by polyethylene glycol, poly-L-omithine, poly-L-arginine or poly-L-histidine.

[0115] The composition for its use according to the invention may be in liquid or solid form. When it is in liquid form, the composition comprises at least water and the constituents mentioned above. The solid form is preferably obtained from the liquid form, preferably by freeze-drying. When the composition is in liquid form and comprises micelles, and is freeze-dried into a solid form, the micelles reform when the solid composition is again placed in an aqueous solution.

[0116] The composition for use according to the invention also preferably comprises at least one pharmaceutically acceptable excipient. The excipient may be chosen in particular to meet the pH and osmolarity requirements of injectable solutions in humans or animals. For example, it may consist of acids or bases to adjust the pH or NaCl to adjust the osmolarity.

[0117] The composition for use according to the invention, in particular composition Cl, is intended for administration to humans or animals and is therefore presented in a form suitable for such administration. When in liquid form, it is preferably suitable for subcutaneous or intravenous administration, particularly intravenous infusion, and is packaged in suitable containers known to those skilled in the art for packaging this type of product. The composition can also be administered in liquid form via a pump, such as an insulin pump.

[0118] When in solid form, it is preferentially suited for administration: - by transcutaneous route and it is preferably formulated and packaged in the form of a patch, or - via nasal application in powder form, or - sublingually in powder or tablet form, or - by transmucosal route and it is preferably formulated and packaged in the form of a mucoadhesive tablet.

[0119] Manufacturing process

[0120] The useful composition according to the invention can be manufactured by any suitable process.

[0121] If the molecules that constitute the composition are used as such in a solvent, they can all be mixed together in said solvent.

[0122] If the molecules in the composition are conjugated to polymers and optionally to micelles for others, the manufacturing process may include the following steps: - a. Create an amphiphilic premix: mix in an aqueous solution: - one or more Butyrate-Poly-L-Lysine conjugates - one or more Lactate-Poly-L-Lysine conjugates - one or more Propionate-Poly-L-Lysine conjugates - b. Optionally, add at least some famesylcysteine ​​to the amphiphilic premix and cholesterol, and stir to form micelles made up of one or more of the conjugates of the amphiphilic premix encapsulating farnesylcysteine ​​and cholesterol.

[0123] Preferably, the manufacturing process may include the following steps: - a. To create an amphiphilic premix: mix in an aqueous solution: - one or more Butyrate-Poly-L-Lysine conjugates - one or more Lactate-Poly-L-Lysine conjugates - one or more Propionate-Poly-L-Lysine conjugates, and at least one or more conjugates chosen from: - one or more Oleyl-Poly-L-Lysine conjugates - one or more Palmitic-Poly-L-Lysine conjugates - one or more Lauryl-Poly-L-Lysine conjugates - one or more Azeayl-Poly-L-Lysine conjugates - one or more Palmitoleyl-Poly-L-Lysine conjugates - one or more Thioctyl-Poly-L-Lysine conjugates - one or more Myristyl-Poly-L-Lysine conjugates - one or several Orotyl-Poly-L-Lysine conjugates - one or more Acetate-Poly-L-Lysine conjugates - one or more Linoleyl-Poly-L-Lysine conjugates, and - b. add at least farnesylcysteine ​​and cholesterol to the amphiphilic premix, and stir to form micelles made up of one or more of the conjugates of the amphiphilic premix encapsulating the farnesylcysteine ​​and cholesterol.

[0124] One embodiment of the invention involves taking advantage of the amphiphilic nature of most of the individual components to create an amphiphilic premix that allows for the controlled solubilization of hydrophobic species. The solubilization process, according to a preferred embodiment, consists of a controlled addition of the hydrophobic molecules to the amphiphilic premixes and allowing sufficient time for their dissolution under stirring.

[0125] Poly-L-lysine can be replaced by another poly-lysine or by polyethylene glycol, poly-L-ornithine, poly-L-arginine or poly-L-histidine.

[0126] Preferably, the agitation is carried out for at least 60 minutes, even more preferably between 5 and 20 minutes, and preferably at an agitation speed less than or equal to 900 revolutions per minute, in particular at an agitation speed between 50 and 800 revolutions per minute.

[0127] According to a preferred embodiment, the manufacturing process according to the invention also includes a step c. of separating the soluble and insoluble phases, to recover the soluble phase. In this case, the insoluble phase is eliminated and the soluble phase constitutes the composition according to the invention. Indeed, a separation process physical (filtration, ultrafiltration) is preferentially carried out to ensure the isolation of the soluble fraction, containing both the amphiphilic premix and the solubilized molecules.

[0128] The composition in liquid form can then be lyophilized or dehydrated to be in solid form, preferably by slow lyophilization for example between 12 and 36 hours.

[0129] Furthermore, if active molecule-polymer conjugates of the composition are not integrated into the premix of step a, these can be added to the mixture after step b, i.e. after the formation of the micelles and the encapsulation of farnesyl-cysteine ​​and cholesterol.

[0130] Active molecule-polymer conjugates can be manufactured by any means known to those skilled in the art for covalently conjugating a molecule to a polymer according to their chemical nature. Thus, lauric acid, myristic acid, palmitic acid, oleic acid, linoleic acid, orotic acid, azelaic acid, thioctic acid, acetic acid, palmitoleic acid, butyric acid, lactic acid, propionic acid, and oleic acid are conjugated to the polymer (preferably poly-L-Lysine) by an amide bond.

[0131] Examples of the realization of amide, urea and carbamate bonds are described, for example, in: -Ryser HJ, Shen WC. Conjugation of methotrexate to poly (L-lysine) as a potential way to overcome drug resistance. Cancer. 1980 Mar 15;45(5 Suppl): 1207-11 (amide bonds), - Zhuxian Z, Jianbin T, Qihang S, William J. A multifunctional PEG-PLL drug conjugale forming redox-responsive nanoparticles for intracellular drug delivery Issue 38, 2015. Journal of Materials Chemistry B (liaisons amide), - Scheper V, Wolf M, Scholl M, Kadlecova Z, Perrier T, Klok HA, Saulnier P, Lenarz T, Stôver T. Potential novel drug carriers for inner ear treatment: hyper-branched polylysine and lipid nanocapsules. Nanomedicine (Lond). 2009 Aug;4(6):623-35 (liaisons urée), - Stéphanie Gac-Breton, Jean Coudane, Mahfoud Boustta & Michel Vert (2004) Norfloxacin-Poly(i-Lysine Citramide Imide) Conjugales and Structure-dependence of the Drug Release, Journal of Drug Targeting, 12:5, 297-307, (liaisons carbamate), - Elmore, W. M. (2013). Nanoparticles Stabilized with MPEG-Polylysine Carbamate: Synthesis and Characterization, (liaisons carbamate), - Ning-Ping Huang, Janos Vôrôs, Susan M. De Paul, Marcus Textor, and Nicholas D. Spencer. Biotin-Derivatized Poly(l-lysine)-g-poly(ethylene glycol): A Novel Polymeric Interface for Bioaffinity Sensing. Langmuir 2002 18 (1), 220-230 (carbamate bonds). Examples

[0132] Example of a composition suitable for use according to the invention:

[0133] Example 1

[0134] An example of a suitable composition according to the invention is presented below.

[0135] [Tables 1] Conjugated PLL (PLL = Poly-L-Lysine) Concentration (mg / mL) Butyrate-PLL 1.16 Lactate-PLL 0.73 Propionate-PLL 0.72

[0136] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0137] Example 2

[0138] An example of a suitable composition according to the invention is presented below.

[0139] [Tables2] Conjugated PLL (PLL = Poly-L-Lysine) Concentration (mg / mL) Butyrate-PLL 0.89 Lactate-PLL 0.56 Propionate-PLL 0.55

[0140] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0141] Example 3

[0142] An example of a suitable composition according to the invention is presented below.

[0143] [Tables3] Conjugated PLL (PLL = Poly-L-Lysine) Concentration (mg / mL) Butyrate-PLL 1.59 Lactate-PLL 1 Propionate-PLL 0.98

[0144] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0145] Example 4

[0146] An example of a composition suitable for testing on mice is shown below.

[0147] [Tables4] Conjugated PLL (PLL = Poly-L-Lysine) Concentration (mg / mL) Butyrate-PLL 0.089 Lactate-PLL 0.056 Propionate-PLL 0.055

[0148] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0149] Example 5

[0150] An example of a suitable composition according to the invention is presented below.

[0151] [Tables5] Conjugated PLL (PLL = Poly-L-Lysine) Concentration (mg / mL) Butyrate-PLL 1.16 Lactate-PLL 0.73 Propionate-PLL 0.72 Thioctyl-PLL 0.92

[0152] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0153] Example 6: Example of Cl composition

[0154] An example of a useful Cl composition according to the invention is presented below.

[0155] The following quantities of conjugates were weighed to prepare a 300 ml stock of composition Cl. All conjugates were weighed with an analytical balance in a laminar flow hood.

[0156] [Tableauxô] PLL Conjugate (PLL = Poly-L-Lysine) Concentration IX (mg / mL) Oleyl-PLL 0.82 Palmitic-PLL 0.32 Lauryl-PLL 0.78 Linoleyl-PLL 0.32 Azelayl-PLL 0.31 Palmitoleyl-PLL 0.64 Thioctyl-PLL 0.92 Myristyl-PLL 0.32 Orothyl-PLL 0.3 Acetate-PLL 0.72 Butyrate-PLL 1.16 Lactate-PLL 0.73 Propionate-PLL 0.72

[0157] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0158] In parallel, cholesterol and farnesylcysteine ​​molecules were dissolved separately at an approximate concentration of 50 mg / ml in absolute ethanol and filtered through a 0.22 µm filter. The solvent was evaporated by a rotary evaporator. The required quantities of dry solids were weighed in a laminar flow hood, as shown in Table 3.

[0159] [Tables?] Molecule Concentration IX (mg / mL) Cholesterol-PLL 0.33 Famesylcysteine-PLL 0.32

[0160] In a sterile 500 ml bottle fitted with a magnetic stir bar, 23.8 ml of each previously prepared conjugate solution were added under magnetic stirring (700 rpm). The corresponding amounts of salts (previously lyophilized) at 300% PBS were added to the conjugate solution under magnetic stirring (700 rpm).

[0161] Cholesterol and farnesylcysteine ​​were added and the mixture was left under agitation for 25 hours (700 revolutions per minute).

[0162] 30 ml of formulation Cl 10 x were taken in a laminar flow hood and added to a new sterile 500 ml bottle.

[0163] 270 ml of sterile PBS were added to obtain formulation Cl Ix. The for The mulation was agitated for 30 minutes.

[0164] Example 7: Example of Cl composition

[0165] An example of a Cl composition suitable for testing on mice is shown below.

[0166] The following quantities of conjugates were weighed to prepare a 300 ml stock of composition CL. All conjugates were weighed with an analytical balance in a laminar flow hood.

[0167] [Tables8] PLL Conjugate (PLL = Poly-L-Lysine) IX Concentration (mg / mL) 10X Concentration (mg / mL) Oleyl-PLL 0.063 0.63 Palmitic-PLL 0.024 0.24 Lauryl-PLL 0.059 0.59 Linoleyl-PLL 0.025 0.25 Azelayl-PLL 0.024 0.24 Palmitoleyl-PLL 0.049 0.49 Thioctyl-PLL 0.07 0.7 Myristyl-PLL 0.024 0.24 Orothyl-PLL 0.023 0.23 Acetate-PLL 0.055 0.55 Butyrate-PLL 0.089 0.89 Lactate-PLL 0.056 0.56 Propionate-PLL 0.055 0.55

[0168] Each conjugate was weighed in a sterile 50 ml centrifuge tube and dissolved in 30 ml of water using a vortex mixer. Each conjugate was shaken for a period of 10 to 20 minutes depending on the solubility of the conjugate.

[0169] In parallel, cholesterol and farnesylcysteine ​​molecules were dissolved separately at an approximate concentration of 50 mg / ml in absolute ethanol and filtered through a 0.22 µm filter. The solvent was evaporated by a rotary evaporator. The required quantities of dry solids were weighed in a laminar flow hood.

[0170] [Tables9] Molecule Concentration IX (mg / mL) Concentration 10X (mg / mL) Cholesterol-PLL 0.004 0.04 Famesylcysteine-PLL 0.004 0.04

[0171] In a sterile 500 ml bottle fitted with a magnetic stir bar, 23.8 ml of each previously prepared conjugate solution were added under magnetic stirring (700 rpm). The corresponding amounts of salts (previously lyophilized) at 300% PBS were added to the conjugate solution under magnetic stirring (700 rpm).

[0172] Cholesterol and farnesylcysteine ​​were added and the mixture was left under stirring for 25 hours (700 revolutions per minute).

[0173] 30 ml of formulation Cl 10 x were taken in a laminar flow hood and added to a new sterile 500 ml bottle.

[0174] 270 ml of sterile PBS were added to obtain formulation Cl Ix. The for The mulation was agitated for 30 minutes.

[0175] The vials were prepared according to the following plan using a sterile, calibrated 5 ml pipette: Cl IX = 83 vials, 2 ml each. Cl 10X = 83 vials of 2 ml each.

[0176] The 166 vials were placed in a steel freeze-drying tank and a one-day freeze-drying cycle was started.

[0177] Example 8: Molecular Analysis of Mouse Fecal Microbiota (Wild Wild WT, SOD1, SOD1 + composition according to the invention).

[0178] The SOD1 mouse is an animal model of axonal degeneration consisting of transgenic mice expressing the mutated form of the human superoxide dismutase gene. It is the reference in vivo model for studying amyotrophic lateral sclerosis.

[0179] In this test, the compositions Cl (described in Tables 8 and 9) and C2 (described in Table 10) were injected one after the other with a 1-hour interval, starting with composition CL [Tables 10] Conjugated PLL (PLL = Poly-L-Lysine) Concentration IX (mg / mL) Concentration 10X (mg / mL) Alpha-tocopherol-PLL 0.070 0.70 Ascorbyl-PLL 0.063 0.63 Biotinyl-PLL 0.068 0.68 GABA-PLL 0.126 1.26 Glutathione-PLL 0.074 0.74 Pantothenic acid-PLL 0.068 0.68 Retynoil-PLL 0.202 2.02 Sperminin-PLL 0.066 0.66 Taurine-PLL 0.110 1.10 PLL / Cysteine ​​(copolymer) 0.06 0.6 PLL / Methionine (copolymer) 0.101 1.01

[0180] Feces from five male and female mice from each group (WT, SOD1, SOD1 + Cl low dose and SOD1 + Cl high dose), collected in the cages, were analyzed at T0 (=T6), T3 weeks (T9) and 10 weeks (T16) for a total of 55 samples.

[0181] In a first step, total genomic DNA was extracted from each fecal sample using the Godon method (Godon et al., Appl Envion. Microbiol., 1997). The quality of the extracted DNA was assessed after migrating the samples onto an agarose gel. The DNA concentration was then determined using Nanodrop technology.

[0182] The DNA samples obtained were then sequenced. The approach used was high-throughput sequencing of the gene encoding 16S ribosomal RNA (V3-V4 region) using Illumina technology (read length: 150 bases, read depth: 1 million reads).

[0183] The taxonomic affiliation of each bacterial sequence obtained (or OTU) was carried out using the databases silva_nr99_vl38.1 (https: / / www.arb-silva.de / ) and GTDB v202 (https: / / gtdb.ecogenomic.org / ).

[0184] The ranomaly pipeline (Theil S and Rifa E. rANOMALY: AmplicoN wOrkflow for Microbial community AnaLYsis [version 1; peer review: 2 approved]. FlOOOResearch 2021, 10:7 https: / / doi.Org / 10.12688 / fl000research.27268.l), based on the dada2 program, was used to process all sequences, both for estimating bacterial phyla / family abundance and for performing statistical analyses. (composition, diversity analyses, differential abundance analyses).

[0185] The comparison of bacterial diversity and richness between microbiomes from treated and untreated mice was carried out by calculating the Shannon index. Differences in population structure (beta diversity) were highlighted by two multivariate analysis methods (ANOVA and Nonmetric Multidimensional Scaling = Method MDS) and a PLS-DA (partial least squares discriminant regression analysis) from the mixOmics package.

[0186] Differential analyses of OTU abundances were performed using the normalization and estimation tools of the DESeq2 package [Love, ML, Huber, W., Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 Genome Biology 15(12):550; 2014].

[0187] Microbiota-Clinical Parameter Correlation

[0188] The potential correlation between each of the clinical and physiological parameters measured and the molecular composition of the microbiota was studied for each of the female mice whose microbiota was analyzed (5 mice per group).

[0189] Several analysis methods were tested, based on rank correlation (spearman).

[0190] The method used is canonical correlation analysis (RCCA) as proposed by the mixOmics package (Rohart F, Gautier B, Singh A, and Le Cao KA, 2017: mixOmics: An R package for 'omics feature selection and multiple data integration. PLoS computational biology 13(11):el005752). This method allows for cross-validation of the results of correlation analyses between OTU abundance and the various clinical parameters measured. The results obtained are presented as heatmaps showing significant correlations, positive or negative, between OTU abundance and the clinical parameters.

[0191] Results:

[0192] Comparison of the composition of the fecal microbiota of "SOD1" KO mice to that of "Wild Type" (WT) mice

[0193] Analysis of the relative abundance of bacterial phyla, orders, families and even genera present in SDO1 mice compared to WT mice shows that the composition of the SOD1 microbiota differs from that of WT:

[0194] - SOD1 have fewer Firmicutes and more Bacteroidetes than WT

[0195] - Among the Bacteroidetes, SOD1 mice host more Bacteroidales in per particular Muribaculaceae and more Porphyromonaceae including Muribaculum than the WT

[0196] - Among the Firmicutes, the relative abundance of the Lachnospiraceae (Shaedlerella and Acetifractor) and Erysipelotrichales is decreased while that of Lactobacillales such as Limolactobacillus and Oscillospiraceae as well as that of Eggerthelaceae is increased compared to WT mice.

[0197] Analysis of the diversity (alpha diversity) and richness (beta diversity) of the fecal microbiota of the SOD1 mouse compared to that of the WT mouse revealed that there are:

[0198] - No significant difference in bacterial diversity (indices: Chaol, Observed, Shannon and Inv Simpson) between SOD1 and WT (alpha diversity).

[0199] - A significant difference (Jaccard: p=0.01 and Bray Curtis: p=0.01) in the Bacterial species richness (beta diversity). No effect of time (T0, T6 and T16) was observed.

[0200] The Jaccard and Bray Curtis index or distance are two well-known metrics for measuring similarity, dissimilarity, and diversity among several samples.

[0201] Finally, PLS-DA analysis shows that the fecal microbiomes of WT and SOD1 mice differentiate ([Fig.1]).

[0202] In conclusion, SOD1 mice have a disturbed gut microbiota compared to wild WT mice, characterized by lower bacterial richness, a higher proportion of Gram- bacteria (Muribaculaceae and Muribaculum) as well as Gram-+ Lactobacillales and Eggerthelaceae but a lower proportion of Lachnospiraceae and Erysipelotrichales.

[0203] Result: Effect of administering composition Cl (associated with C2) at high or low dose in mice SOD1

[0204] A - Administration of a high 10X dose of Cl (associated with C2)

[0205] The Cl and C2 compositions are described in Tables 7 to 10.

[0206] Administration of the composition Cl (associated with C2) at a high dose for 3 weeks results in: - An increase in the relative abundance of: Bacteroidales especially Muribaculaceae Oscillospiraceae COE1 - A decrease in the abundance of: Lactobacillaceae (Lactobacillus genus) Lachno spiraceae Administering a high dose of the Cl composition for 3 weeks appears to accentuate the disturbances observed in SOD1 mice.

[0207] B - Administration of a low dose IX of Cl (associated with C2)

[0208] Administration of a low dose of Cl (associated with C2) to SOD1 mice also causes changes in the relative abundance of certain microbial groups. Thus, we observe: - An increase in the relative abundance of: Lachno spiraceae GAC-485 Oscillospirals - A decrease in the relative abundance of: Lactobacillus

[0209] The low dose appears to partially rebalance the gut microbiota of SOD1 mice to more closely resemble the composition of the microbiota of wild-type (WT) mice, notably by causing an increase in Lachnospiraceae and a decrease in Lactobacillales. This effect appears to be progressive, as it is more pronounced after 10 weeks of treatment than after 3 weeks.

[0210] Result: Comparison of bacterial diversity and richness in treated and untreated SOD1 mice.

[0211] Analysis of the alpha diversity of fecal microbiota did not reveal a significant effect of administration of Cl (associated with C2), at low and high doses, on bacterial diversity in SOD1 mice. Only a significant difference (Shannon index, p=0.02) was observed between the fecal microbiota of mice treated with the low dose and those treated with the high dose of Cl (associated with C2) ([Fig.2] and 3).

[0212] Beta diversity analysis showed significant differences between untreated and treated SOD1 mice. - Controls versus Cl (associated with C2) high dose (p=0.02 Jaccard and p= 0.01 Bray curtis) - Cl (associated with C2) low dose versus Cl (associated with C2) high dose (p = 0.02 Jaccard and p = 0.01 Bray Curtis) - No significant difference between untreated control and Cl (associated with C2) low dose.

[0213] Results: Comparison between wild-type mice and SOD1 mice treated or not with different doses of Cl (associated with C2)

[0214] PLS-DA analysis of the composition of all the microbiomes studied showed that these microbiomes could be separated into 4 distinct groups, with the microbiota of mice treated with the low dose of Cl (associated with C2) being close, in the same cluster, to the microbiota of wild mice ([Fig.4]).

[0215] Conclusion

[0216] The study showed that the composition of the microbiota of SOD1 mice differs from that of wild-type mice, with these alterations affecting the richness of the microbiota more than its diversity. The microbiota of SOD1 mice appears disrupted, as the relative abundance of certain major bacterial groups is altered. It is characterized, in particular, by a higher abundance of Muribaculaceae and Lactobacillales (Oscillospiraceae) at the expense of Lachnospiraceae and Erysipelotrichales. PLS-DA analysis further demonstrated that the microbiota of wild-type and SOD1 mice were separated.

[0217] Administration of a high dose of Cl (combined with C2) to SOD1 mice for 3 weeks appears to amplify the differences observed between wild-type and SOD1 mice (increased Muribaculaceae and Oscillospiraceae and decreased Lachnospiraceae). The microbiota composition of SOD1 mice treated with the high dose of Cl (combined with C2) is very distinctly different from that of SOD1 controls, but also from that of wild-type mice; this group of mice treated with the high dose differs significantly from the others (most disrupted microbiota).

[0218] Administration of a low dose of Cl (combined with C2) to SOD1 mice for 3 to 10 weeks appears to partially rebalance the microbiota disturbances observed in control SOD1 mice. Specifically, an increase in Lachnopiracea and a decrease in Lactobacillales, microbial groups affected in SOD1 mice compared to wild-type mice, are observed. The microbiota composition of SOD1 mice treated with a low dose of Cl (combined with C2) thus becomes closer to that of wild-type mice, with the microbiota of these two groups clustered together.

[0219] These preliminary results are of interest with regard to the administration of the lowest dose in the SOD1 mouse model. At this dose, the Cl composition (associated with C2) does indeed appear to have a beneficial effect on the disrupted microbiota of SOD1 mice.

[0220] Example 9: Correlation study between symptoms and molecular composition of the microbiota in SOD1 mice after administration of a low dose of Cl (associated with C2)

[0221] The objective of the study was to analyze the potential correlations between the molecular composition of the gut microbiota and clinical parameters measured in SOD1 mice versus wild type mice and in SOD1 mice treated or not with a low dose of Cl (associated with C2) for 10 weeks.

[0222] This study was carried out in female mice whose fecal microbiota had been previously molecularly analyzed (five females from each group [WT, SOD1, SOD1- low dose], collected from the cages, at T0 (=T6), T3 weeks (T9) and 10 weeks (T16)).

[0223] The tests were performed at least 1 hour after the first daily dose of test compound or vehicle.

[0224] A one-day session included a 5-minute training trial at 4 RPM on the rotary apparatus (AccuScan Instruments, Columbus, USA). One hour later, the animals were tested in three consecutive 6-minute acceleration trials with the speed increasing from 0 to 40 RPM over 360 s and an inter-trial interval of at least 30 minutes. The latency to fall from the rod was recorded.

[0225] Correlation study in SOD1 and Wild Type mice,

[0226] The RCCA (Regularized Canonical Correlation Analysis) primarily demonstrated, in WT and SOD1 mice, a significant negative correlation between clinical score, distance traveled, and recovery time after Rotarod and the Desulfovibrionaceae family, as well as a weaker negative correlation with the Prevotellaceae family. Thus, the higher the abundance of these bacterial families, the lower the clinical score, as well as the distance traveled and recovery time ([Fig. 5]).

[0227] Although the relative abundance of the Desulfovibrionaceae family (like that of the Prevotellaceae) is low compared to other bacterial families, we were nevertheless able to detect OTUs from this family only in SOD1 mice (no detection in Wild Type mice). This could suggest a potential role for Desulfovibrionaceae in the pathophysiology of SOD1 mice.

[0228] Correlation study in SOD1 mice treated or untreated with the low dose of composition Cl (associated with C2) for 10 weeks

[0229] RCCA correlation analysis performed in control SOD1 mice treated with the low dose of PL revealed a strong positive correlation between the Lachnospiraceae family and the distance traveled by the mice. The more abundant this family, the greater the distance traveled by the mice. Given that one of the main effects of a composition according to the invention on the gut microbiota of SOD1 mice is to increase the abundance of Lachnospiraceae species, the observed correlation suggests a role for this bacterial family in restoring the ability of SOD1 mice to travel a distance close to that traveled by Wild Type mice ([Fig. 6]).

[0230] The demonstration of a negative correlation between Desulfovibrionaceae and the severity of symptoms in SOD1 mice suggests that sulfide-producing species in this family may be involved in the pathophysiology, as has been observed in other pathologies such as irritable bowel syndrome or chronic inflammatory bowel diseases (Crohn's disease, ulcerative colitis, etc.). Concurrently, the lower abundance of Lachnospiraceae species in SOD1 mice could result in a decrease in intracolonic butyrate concentration, as a large proportion of Lachnospiraceae species produce this metabolite with demonstrated health benefits.

[0231] Administration of a low-dose composition according to the invention for 10 weeks is accompanied by changes in the composition of the SOD1 mouse microbiota, resulting, in particular, in an increase in the abundance of Lachnospiraceae and the disappearance of Desulfovibrionaceae. This correlates with the significant improvement in the mice's ability to travel a distance equivalent to that of Wild Type mice. The administration of a composition according to the invention would therefore make it possible both to partially rebalance the altered microbiota of the SOD1 mouse, leading to metabolic changes and to reduce the severity of certain symptoms in the SOD1 mouse or even to restore a normal phenotype.

Claims

Demands

1. Composition comprising at least: - lactic acid, and / or a salt and / or an ester and / or anhydride of lactic acid, - butyric acid, and / or a salt and / or an ester and / or anhydride of butyric acid - propionic acid, and / or a salt and / or an ester and / or anhydride of propionic acid, for its use in humans or animals in the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

2. Composition for its use according to the preceding claim, said prevention and / or said treatment being characterized by an increase in the proportion of butyrate-producing bacteria and / or a decrease in the proportion of sulfide-producing bacteria, in the intestinal microbiota.

3. Composition for its use according to claim 1 or 2, characterized in that the pathological dysbiosis of the intestinal microbiota is characterized by an excess of sulfide-producing bacteria and a deficiency of butyrate-producing bacteria.

4. Composition for its use according to the preceding claim, characterized in that pathological dysbiosis of the intestinal microbiota is also characterized by an increase in intestinal inflammation.

5. Composition for its use according to any one of claims 1 to 4, in the prevention and / or treatment of at least one neurodegenerative disease and / or bowel disease associated with pathological dysbiosis of the intestinal microbiota.

6. Composition for its use according to claim 5, characterized in that the neurodegenerative disease is selected from Charcot's disease, amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease.

7. Composition for its use according to claim 5, characterized in that the bowel disease is selected from Crohn's disease, chronic inflammatory bowel disease, ulcerative colitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis and gluten intolerance.

8. Composition according to its use according to one of the preceding re- indications, as a medicine or food supplement for humans or animals.

9. Composition for its use according to any one of claims 2 to 8, characterized in that the sulfide-producing bacteria belong to the Desulfovivrionaceae family and the butyrate-producing bacteria belong to the Lachnospiraceae family.

10. Composition for its use according to any one of claims 8 or 9, characterized in that bacteria belonging to the Desulfo-vibrionaceae family represent less than 0.01% of the total bacteria present in the intestinal microbiota.

11. Composition for its use according to any one of the preceding claims, characterized in that the composition comprises at least one polymer selected from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

12. Composition for use according to any one of the preceding claims, characterized in that at least one of the molecules of the composition selected from lactic acid, butyric acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer selected from poly-lysine, polyethylene glycol, polyomithine, polyarginine and polyhistidine.

13. A composition for use according to any one of the preceding claims, characterized in that the composition also comprises at least one molecule selected from: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid, - acetic acid, and combinations thereof, said molecule(s) being in the form of a salt and / or a ester and / or an anhydride of one or more of these molecules.

14. Composition for its use according to any one of the preceding claims, characterized in that the composition comprises at least the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesyl cysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid - acetic acid, and their combination, and / or a salt and / or an ester and / or anhydride of one or more of these molecules.

15. Composition for use according to any one of the preceding claims, characterized in that at least one of the molecules of the composition selected from oleic acid, palmitic acid, lauric acid, linoleic acid, azelaic acid, palmitoleic acid, thioctic acid, myristic acid, orotic acid, acetic acid, butyric acid, lactic acid, propionic acid, salts of these acids, esters of these acids and anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer selected from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

16. Composition according to any one of claims 12 to 15, characterized in that it comprises micelles in which are encapsulated at least farnesyl cysteine ​​and / or cholesterol and / or an ester of these molecules.

17. Composition according to the preceding claim, characterized in that at least one of the micelles is formed by amphiphilic conjugates, each consisting of at least one hydrophobic molecule covalently conjugated to a molecule of a polymer selected from poly-lysine, polyethylene glycol, polyomithine, polyarginine, and polyhistidine.

18. Composition according to the preceding claim, characterized in that at least one micelle is formed by amphiphilic conjugates each consisting of at least one molecule selected from oleic acid, palmitic acid, lauric acid, linoleic acid, palmitoleic acid, myristic acid, salts, esters and anhydrides of these fatty acids, covalently conjugated to a molecule of a polymer selected from poly-lysine, polyethylene glycol, polyornithine, polyarginine and polyhistidine.

19. Composition for use according to any one of the preceding claims, the composition being characterized in that at least one of the molecules is covalently conjugated to at least one polymer.

20. Composition for use according to any one of the preceding claims, the composition being characterized in that it comprises at least: - A. the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine: - one or more Oleyl-Poly-L-Lysine conjugates - one or more Palmitic-Poly-L-Lysine conjugates - one or more Lauryl-Poly-L-Lysine conjugates - one or more Azelayl-Poly-L-Lysine conjugates - one or more Palmitoleyl-Poly-L-Lysine conjugates - one or more Thioctyl-Poly-L-Lysine conjugates - one or more Myristyl-Poly-L-Lysine conjugates - one or more Orotyl-Poly-L-Lysine conjugates - one or more conjugates Acetate-Poly-L-Lysine - one or more conjugates Butyrate-Poly-L-Lysine - one or more conjugates Lactate-Poly-L-Lysine - one or more conjugates Propionate-Poly-L-Lysine, - one or more conjugates linoleyl-Poly-L-Lysine, and - B.famesylcysteine ​​and cholesterol, and / or an ester of these molecules, encapsulated in micelles.

21. Composition for its use according to the preceding claim, the composition being characterized in that famesylcysteine ​​and cholesterol, and / or an ester of these molecules, are encapsulated in micelles formed by one or more of the conjugates of list A.

22. Composition for its use according to claim 20 or 21, the composition being characterized in that Poly-L-Lysine is replaced by another poly-lysine or by polyethylene glycol, poly-L-omithine, poly-L-arginine or poly-L-histidine.

23. Composition for its use according to any one of the preceding claims, characterized in that the composition comprises at least one pharmaceutically acceptable excipient.

24. Composition for use according to any one of the preceding claims, characterized in that the composition is in liquid or solid form.

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