Composition for preventing and / or treating dysbiosis of the gut microbiota

Abstract

Meeting the need for new therapies for preventing or treating dysbiosis of the gut microbiota, particularly for treating or preventing neurodegenerative or intestinal diseases. The present invention relates to the use of a composition comprising a plurality of specific molecules for treating dysbiosis of the gut microbiota. Dysbiosis of the gut microbiota can be characterized by excessive sulfide-producing bacteria and depleted butyrate-producing bacteria.

Description

Technical Field

[0001] The present invention relates to the prevention and / or treatment of the dysbiosis of the gut microbiota. In particular, the present invention relates to the use of a composition comprising a plurality of specific molecules for treating the dysbiosis of the gut microbiota.

Background Art

[0002] The microbiota is an aggregate of microorganisms (bacteria, archaea, viruses, and eukaryotes) that propagate in a specific environment.

[0003] The human body contains a large number of microbiota, especially on the skin, in the mouth, vaginal cavity, throat, and digestive system.

[0004] The microbiota present in the digestive system is also known as the gut microbiota, which is distributed from the stomach to the colon and occupies 35-50% of the volume of the intestinal contents relative to a weight exceeding 1 kilogram.

[0005] The gut microbiota is home to most of the 10 trillion microorganisms that inhabit the human body.

[0006] Although discovered in the 19th century, scientists were unable to identify all the genes in the gut microbiota until the advent of high-throughput sequencing in the 2000s. Characterizing all the microbial genomes found in the gut has led to the discovery of more than 1000 different species, the majority of which are bacteria.

[0007] A detailed analysis of the gut microbiota shows that, like fingerprints, the gut microbiota is unique to each individual and depends on their environmental and genetic properties.

[0008] Once formed, the composition of the microbiota remains relatively stable throughout an individual's life. The variation is generally less than 5% of the total microbiota.

[0009] As a result, pathogenic conditions in the gut microbiota are difficult to detect because they are derived from these specific species.

[0010] When a patient presents with a disease, an imbalance in the pathogenic gut microbiota, known as dysbiosis, may be found. Without the presence of biomarkers for the disease, it is difficult to establish thresholds for the imbalance, similar to the variations in genera and species.

[0011] Thanks to the removal of technical barriers, the analysis of the gut microbiota has revealed its major role in both maintaining good health of the body and the development of many diseases.

[0012] The gut microbiota plays an important role in the body's functions. The microorganisms that make it up have a wide range of actions, from food decomposition to epithelial repair and metabolic regulation.

[0013] Nevertheless, through dysbiosis, i.e., changes in the composition of the microbiota, the microbiota can be associated with induced or triggered pathological disorders such as neurodegeneration and intestinal diseases as well as certain cancers.

[0014] Central nervous system - gut interactions are a major factor in the development of certain pathological conditions. In Parkinson's disease, the association between dysbiosis of the gut microbiota and the development of motor disorders and neuroinflammation has been demonstrated.

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

[0016] Furthermore, it is now known that intestinal diseases, more specifically, inflammatory bowel disease, Crohn's disease, and food intolerances, particularly gluten intolerance, are associated with the presence of a gut microbiota depleted in microorganisms and thus pathological dysbiosis.

[0017] Currently, the solutions proposed to prevent or control dysbiosis essentially consist of administering probiotics and / or prebiotics, particularly for treating intestinal diseases, but their effectiveness is highly variable and depends on the patient's basic gut microbiota.

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

[0019] Furthermore, especially with regard to degenerative diseases, these are often very difficult for patients. Some symptoms can be alleviated, but such diseases are never completely cured, and there is currently no satisfactory treatment. Many chemical agents have therapeutic properties, but due to their toxicity, their lifespan, or their rapid elimination, they are not suitable for treating degenerative diseases.

[0020] Specifically, despite being multifactorial, current treatments target only one of the causative factors of these diseases and do not attempt to act more generally using a systemic approach that directly targets the cause of the disease. For example, Lou Gehrig's disease or amyotrophic lateral sclerosis is associated with a number of factors, particularly oxidative stress, mitochondrial dysfunction, neuroinflammation, excitotoxicity, oligodendrocyte dysfunction and degeneration, changes in proteostasis, changes in the DNA repair system, changes in nuclear cytoplasmic RNA and RNA associated with transport proteins, defects in axonal transport, and defects in vesicular transport, as well as pathological dysbiosis of the gut microbiota. Currently, the treatment recommended for amyotrophic lateral sclerosis is riluzole®, which acts only on the inhibition of glutamate release to counter excitotoxicity and does not consider the effect on the gut microbiota. The same observation can be made for other degenerative diseases.

[0021] Therefore, there is a strong need for solutions for preventing or treating the dysbiosis of the gut microbiota, in particular for treating or preventing neurodegenerative or intestinal diseases, and for meeting the essential requirements for new therapies.

[0022] Therefore, the object of the present invention is to meet all of these needs and overcome the disadvantages and limitations of the prior art.

Summary of the Invention

[0023] To achieve this object, the present invention proposes the use of a specific composition for preventing and / or treating the dysbiosis of the gut microbiota, which is suitable for use in the treatment or prevention of diseases associated with the dysbiosis of the gut microbiota, in particular neurodegenerative diseases or intestinal diseases.

[0024] For this purpose, the present invention relates to a composition comprising at least - lactic acid, and / or salts and / or esters and / or anhydrides of lactic acid - butyric acid, and / or salts and / or esters and / or anhydrides of butyric acid, and - propionic acid, and / or salts and / or esters and / or anhydrides of propionic acid for use in humans or animals in the prevention and / or treatment of the dysbiosis of the gut microbiota.

[0025] Advantageously, the composition according to the present invention can restore the balance of the microbiota of an individual presenting dysbiosis of the gut microbiota.

[0026] Preferably, the composition according to the present invention is used to treat the dysbiosis of the gut microbiota characterized by excessive sulfide-producing bacteria and depleted butyrate-producing bacteria in the gut microbiota.

[0027] Accordingly, the treatment according to the invention is preferably 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.

[0028] Advantageously, the effects on butyrate-producing bacteria and sulfide-producing bacteria provide a synergistic action for treating dysbiosis, in particular dysbiosis associated with a pathological condition.

[0029] 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 intestinal disease associated with dysbiosis of the intestinal microbiota. According to a particularly preferred variant, the composition can be used in humans or animals for the prevention and / or treatment of at least one neurodegenerative disease and / or intestinal disease associated with dysbiosis of the intestinal microbiota, and the human or animal exhibits intestinal dysbiosis characterized in the intestinal microbiota by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria.

[0030] Surprisingly, the inventors have found that the composition according to the invention can treat or prevent diseases associated with dysbiosis, in particular neurodegenerative diseases or intestinal diseases associated with dysbiosis, such as amyotrophic lateral sclerosis or intestinal diseases such as Crohn's disease.

[0031] The composition according to the invention can be used as a drug or dietary supplement for humans or animals.

[0032] According to one variant, the composition according to the invention can be used for the prevention or treatment of neurodegenerative diseases associated with dysbiosis of the intestinal microbiota selected from amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease.

[0033] According to another variant, the composition according to the invention can be used for preventing or treating intestinal diseases associated with a pathological dysbiosis of the gut microbiota selected from Crohn's disease, chronic inflammatory bowel disease, hemorrhagic proctocolitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis, and food intolerances, in particular gluten intolerance.

[0034] According to a particularly preferred embodiment, the composition for use according to the invention, in addition to lactic acid, butyric acid, and propionic acid, and / or salts and / or esters and / or anhydrides of these molecules, - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesylcysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - acetic acid, may also contain at least one molecule selected from combinations thereof, the molecule(s) being potentially in the form of salts and / or esters and / or anhydrides of one or more of these molecules.

[0035] Preferably, the composition for its use is the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesylcysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - acetic acid, - butyric acid, - lactic acid, - propionic acid, and / or one or more salts and / or esters and / or anhydrides of these molecules.

[0036] Advantageously, these molecules each act on different factors, enabling a multi-factorial action on the gut microbiota. These molecules help to balance excessive or depleted bacterial populations, particularly by reducing intestinal inflammation.

[0037] Advantageously, the molecules of the composition for use according to the invention are conjugated to a specific polymer such as polylysine, thereby enhancing the effectiveness and bioavailability of these molecules.

[0038] Thus, according to a particularly preferred embodiment, the composition for use according to the invention comprises at least the following conjugates, each conjugate consisting of a molecule covalently bound to polylysine. - one or more butyrate-poly-L-lysine conjugates, - one or more lactate-poly-L-lysine conjugates, - one or more propionate-poly-L-lysine conjugates.

[0039] According to one variant form, the composition for use according to the invention comprises at least - A. the following conjugates, each conjugate consisting of a molecule covalently bound to polylysine: - one or more oleyl-poly-L-lysine conjugates, - one or more palmitin-poly-L-lysine conjugates, - one or more lauryl-poly-L-lysine conjugates, - one or more azelail-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. micelles, preferably micelles formed by one or more of the conjugates listed in List A, encapsulating farnesyl cysteine and cholesterol, and / or esters of these molecules.

[0040] The present invention also relates to a method for producing these compositions.

[0041] Other features and advantages will become apparent from the detailed description of the invention and the following examples.

Brief Description of the Drawings

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DETAILED DESCRIPTION OF THE INVENTION

[0043] Definitions For the purposes of the present invention, the term "animal" means any animal other than a human. In particular, it is a mammal.

[0044] For the purposes of the present invention, the term "amphiphilic conjugate" means a conjugate formed by a hydrophobic molecule X and a hydrophilic polymer Y, and thus the conjugate has amphiphilic characteristics.

[0045] For the purposes of the present invention, the term "molecule X conjugated to polymer Y" means molecule X covalently bonded to polymer Y, and this bond is preferably an amide, urea, or carbamate bond, depending on the chemical properties of molecule X and polymer Y.

[0046] Within the scope of the present invention, the terms "medical nutrition composition" or "medical nutrition product" or "medical food" or "food for special medical purposes" (FSMP) or "dietary food for special medical purposes" (DFSMP) are construed to mean foods having a therapeutic purpose of prevention and / or treatment, used alone or in combination with other therapies. It is a nutritional compound adapted to a specific clinical situation and is likely to constitute an exclusive or partial diet for the intended patient.

[0047] Within the scope of the present invention, the term "dysbiosis" means a pathological state characterized by an imbalance in the distribution of bacteria in the gut microbiota at the phylum, class, order, family, genus, or species level, which poses a risk of other diseases of metabolic origin.

[0048] For the purposes of the present invention, the term "drug" refers to a product that has received marketing approval as a prophylactic and / or therapeutic agent for a disease.

[0049] Within the scope of the meaning of the present invention, the term "microbiota" is understood to mean the gut microbiota.

[0050] Within the scope of the meaning of the present invention, the term "butyrate-producing bacteria" means bacteria that can produce butyrate by anaerobic fermentation of non-digestible dietary fiber. Butyrate-producing bacteria mainly belong to the phylum Firmicutes.

[0051] Within the scope of the meaning of the present invention, the term "sulfide-producing bacteria" means bacteria that can reduce sulfate to sulfide.

[0052] Within the scope of the meaning of the present invention, the term "alpha diversity" means the measurement of the number of species present in a given sample.

[0053] Within the scope of the meaning of the present invention, the term "beta diversity" means the measurement of the diversity of species in a given sample.

[0054] Within the scope of the meaning of the present invention, the term "excessive sulfide-producing bacteria" means a higher proportion of sulfide-producing bacteria than normal, preferably higher than the proportion of sulfide-producing bacteria in individuals without pathological dysbiosis.

[0055] Within the scope of the meaning of the present invention, the term "deficient butyrate-producing bacteria" means a lower proportion of butyrate-producing bacteria than normal, preferably lower than the proportion of butyrate-producing bacteria in individuals without pathological dysbiosis.

[0056] Compositions for its use Thus, the present invention relates to at least - lactic acid, and / or salts and / or esters and / or anhydrides of lactic acid - butyric acid, and / or salts and / or esters and / or anhydrides of butyric acid, and - A composition comprising propionic acid, and / or a salt and / or ester and / or anhydride of propionic acid for use in humans or animals for the prevention and / or treatment of dysbiosis of the gut microbiota.

[0057] Preferably, the composition is particularly suitable for use in humans for the prevention and / or treatment of dysbiosis of the gut microbiota.

[0058] Advantageously, the combination of these three molecules ensures a significant preventive and / or therapeutic effect against dysbiosis of the gut microbiota. Lactic acid, butyrate, and propionic acid are the main short-chain fatty acids produced when fiber is broken down by bacteria in the gut microbiota. These three fatty acids are involved in many cellular functions and are the main energy source for the intestinal epithelium, more specifically, the epithelial cells of the small intestine and colon. If the content of short-chain fatty acids is too low, it may cause dysbiosis.

[0059] In the context of the present invention, dysbiosis of the gut microbiota is preferably characterized by excessive sulfide-producing bacteria and depleted butyrate-producing bacteria.

[0060] The presence of excessive sulfide-producing bacteria is a characteristic of dysbiosis. In fact, as the proportion of beneficial microorganisms in the microbiota decreases, a bacterial population that has harmful effects or produces metabolites harmful to organisms appears. Thus, patients with dysbiosis have a high level of sulfur-reducing bacteria associated with an increased production of toxic substances such as hydrogen sulfide, which is known for its pro-inflammatory activity. The presence of these metabolites causes inflammation and increased intestinal permeability.

[0061] Thus, the use of the composition according to the present invention is characterized by an increase in the proportion of butyrate-producing bacteria and a decrease in the proportion of sulfide-producing bacteria in the gut microbiota.

[0062] Advantageously, reducing the proportion of sulfide-producing bacteria reduces intestinal inflammation by treating dysbiosis. On the other hand, increasing the proportion of butyrate-producing bacteria allows the intestine and colon epithelial cells to restore membrane permeability. This increase stimulates the microbiota, particularly beneficial bacterial species for the body. Thus, the combined action on butyrate-producing and sulfide-producing bacteria provides a synergistic effect against the dysbiosis of the gut microbiota. The present composition guarantees a long-term effect on the gut microbiota by targeting the cause of dysbiosis.

[0063] The composition can be used to prevent or treat any disease that has led to, or is caused by, a dysbiosis of the gut microbiota, preferably a dysbiosis of the gut microbiota characterized by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria, and preferably also characterized by an increase in intestinal inflammation.

[0064] More specifically, at least - lactic acid, and / or salts and / or esters and / or anhydrides of lactic acid - butyric acid, and / or salts and / or esters and / or anhydrides of butyric acid, and - propionic acid, and / or salts and / or esters and / or anhydrides of propionic acid, the composition can be used in humans or animals for the prevention and / or treatment of at least one disease associated with dysbiosis of the gut microbiota, in particular, the human or animal has an intestinal dysbiosis characterized by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria, in particular, at least one neurodegenerative disease and / or intestinal disease associated with dysbiosis of the gut microbiota, for example, the human or animal has at least one neurodegenerative disease and / or intestinal disease associated with dysbiosis of the gut microbiota, and the human or animal has an intestinal dysbiosis characterized by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria in the microbiota.

[0065] In the context of the present invention, the neurodegenerative disease may be selected from multiple sclerosis, amyotrophic lateral sclerosis (also known as Lou Gehrig's disease), Parkinson's disease, and Alzheimer's disease.

[0066] Furthermore, the intestinal disease may be selected from Crohn's disease, chronic inflammatory bowel disease, hemorrhagic proctocolitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis, and food intolerance, particularly gluten intolerance.

[0067] According to one variant form, the composition can be used to treat or prevent autism spectrum disorder or spondyloarthritis. In particular, a human or animal body has intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfide-producing bacteria and a decrease in the proportion of butyrate-producing bacteria.

[0068] Thus, the composition treats pathological dysbiosis, in particular in combination with a decrease in the proportion of sulfide-producing bacteria, by relatively increasing the proportion of butyrate-producing bacteria in the intestinal microbiota.

[0069] Thus, the composition acts on the intestinal epithelium by providing short-chain fatty acids such as butyric acid that are essential for the function of epithelial cells. Epithelial cells act on inflammation and treat pathological dysbiosis by promoting the growth of butyrate-producing bacteria while restoring membrane permeability.

[0070] Thus, the composition can be used as a drug, a medical nutritional product, or a dietary supplement to restore the balance of the intestinal microbiota by increasing the proportion of sulfide-producing bacteria and decreasing the proportion of butyrate-producing bacteria, and / or to prevent and address diseases resulting from or caused by this pathological imbalance of the intestinal microbiota.

[0071] In the context of the present invention, the sulfide-producing bacteria can be bacteria belonging to the family Desulfovibrionaceae and / or proteolytic bacteria.

[0072] The sulfide-producing bacteria belonging to the family Desulfovibrionaceae are - Alteridesulfovibrio, - Aminidesulfovibrio, - Bilophila, - Desulfocurvus, - Desulfovibrio, - Frigididesulfovibrio, - Fundidesulfovibrio, - Halodesulfovibrio, - Humidesulfovibrio, - Lawsonia, - Maridesulfovibrio, - Megalodesulfovibrio, - Nitratidesulfovibrio, - Oleidesulfovibrio, - Paradesulfovibrio, - Paucidesulfovibrio, - Pseudodesulfovibrio, may belong to at least one bacterial genus selected from Solidesulfovibrio.

[0073] Preferably, the sulfide-producing bacteria belong to the family Desulfovibrionaceae, the genus Bilophila, and the species Wadworthia.

[0074] Therefore, the composition according to the invention can be used in humans or animals in order to reduce the proportion of the bacterial species Bifidobacterium wadsworthia in humans or animals having an intestinal dysbiosis characterized in the microbiota by an increase in the proportion of sulfide-producing bacteria.

[0075] The composition also - acts on at least one butyrate-producing bacterial family selected from Lachnospiraceae - Ruminococcaceae.

[0076] Butyrate-producing bacteria belonging to the family Lachnospiraceae - may belong to at least one bacterial genus selected from Lachnospira, - Roseburia, - Eubacterium, - Anaerostipes.

[0077] Butyrate-producing bacteria of the family Ruminococcaceae may belong to bacteria of the genus Faecalibacterium.

[0078] In a particularly preferred embodiment, the composition can be used to increase the proportion of bacteria from the family Lachnospiraceae and decrease the proportion of bacteria from the family Desulfovibrionaceae, preferably such that the family Desulfovibrionaceae corresponds to less than 0.01% of the total bacteria present in the intestinal microbiota.

[0079] Advantageously, the composition reduces inflammation of the digestive system by restoring the concentrations of sulfide and butyrate to those of a healthy patient.

[0080] Preferably, the composition, in addition to lactic acid, butyric acid, and propionic acid, and / or salts and / or esters and / or anhydrides of these 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 also includes at least one molecule selected from combinations thereof, The molecule is potentially in the form of one or more salts and / or esters and / or anhydrides of these molecules.

[0081] Preferably, the composition - 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 includes at least two molecules, particularly at least three, and even more preferably at least four molecules selected from combinations thereof, The molecule is potentially in the form of one or more salts and / or esters and / or anhydrides of these molecules.

[0082] According to a particular embodiment of the invention, the composition, in addition to lactic acid, butyric acid, and propionic acid, and / or salts and / or esters and / or anhydrides of these molecules, also includes at least azelaic acid, cholesterol, myristic acid, and lauric acid, and / or salts and / or esters and / or anhydrides of these molecules.

[0083] For all active molecules present in the composition for use according to the invention, if they are mentioned in the present application, they can be the molecule itself (e.g., butyric acid) and / or salts of these molecules (e.g., butyrate) and / or esters and / or anhydrides (e.g., butyric acid ester) and / or anhydrides.

[0084] 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, - Farnesylcysteine, - Palmitoleic acid, - Cholesterol, - Thioctic acid, - Myristic acid, - Orotic acid, - Lactic acid, - Propionic acid, - Butyric acid, - Acetic acid, and combinations thereof, The molecule(s) can potentially be in the form of one or more salts and / or esters and / or anhydrides of these molecules.

[0085] Such a composition can, surprisingly and unexpectedly, act on the intestinal microbiota, in particular, increase the proportion of butyrate-producing bacteria and decrease the proportion of sulfide-producing bacteria.

[0086] The useful composition according to the invention comprises at least the following molecules: - Oleic acid, - Palmitic acid, - Lauric acid, - Linoleic acid, - Azelaic acid, - Farnesylcysteine, - Palmitoleic acid, - Cholesterol, - Thioctic acid, - Myristic acid, - Orotic acid, - Lactic acid, - Propionic acid, - Butyric acid, - Acetic acid, and all combinations thereof, The molecule(s) may potentially be in the form of one or more salts and / or esters and / or anhydrides of these molecules, which is hereinafter referred to as composition C1.

[0087] The useful composition according to the present invention, in particular composition C1, may optionally also contain pyruvic acid and / or salts and / or esters and / or anhydrides.

[0088] Preferably, - In the composition, more specifically, the amount of butyric acid in composition C1 is greater than the amount of each of the other molecules individually ingested, and / or - In the composition, more specifically, the amount of thioctic acid in composition C1 is greater than the amount of each of the other molecules excluding butyric acid, and / or - In the composition, more specifically, the amount of lauric acid in composition C1 is greater than the amount of each of the following molecules individually ingested: palmitic acid, linoleic acid, azelaic acid, farnesylcysteine, palmitoleic acid, cholesterol, myristic acid, and orotic acid, - In the composition, more specifically, in composition C1, the amount of oleic acid on the one hand and the amount of lactic acid on the other hand are greater than the amount of each of the following molecules individually ingested: palmitic acid, lauric acid, linoleic acid, azelaic acid, farnesylcysteine, palmitoleic acid, cholesterol, myristic acid, orotic acid, and acetic acid.

[0089] Preferably, each molecule in composition C1 represents 0.5E - 05M to 10E - 05M.

[0090] According to certain embodiments of the present invention, the useful composition according to the present invention, particularly composition C1, can be used by administering a dose of 0.5 to 10 mg / mL, particularly 2 to 8 mg / mL, preferably 7 mg / mL, to treat the dysbiosis of the gut microbiota.

[0091] According to one variant, the useful composition according to the present invention, particularly composition C1, is administered at a first "attack" dose of 7 to 10 mg / mL, preferably 9 to 10 mg / mL, and subsequently preferably at a "maintenance" dose of 0.5 to 5 mg / mL, particularly 2 to 3 mg / mL.

[0092] Preferably, the useful composition according to the present invention, particularly C1, may comprise at least one polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0093] More specifically, in order to improve the solubility, effectiveness, and bioavailability of the molecules of the useful composition according to the present invention, at least one of the molecules of the composition selected 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 selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0094] When the composition is composition C1 or a composition comprising a molecule from the following list, preferably 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 polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0095] Specifically, 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, esters of these acids, and anhydrides of these acids can be covalently conjugated to a polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine, particularly via an amide bond, a urea bond, or a carbamate bond.

[0096] According to a particularly preferred variant, when 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 are present in the composition according to the invention, they are covalently conjugated to at least one molecule of a polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0097] The useful composition according to the invention may contain micelles. When the composition according to the invention, particularly composition C1, contains one of farnesylcysteine and / or cholesterol and / or their esters and / or salts and / or anhydrides, the useful composition according to the invention preferably contains micelles encapsulating one of farnesylcysteine and / or cholesterol and / or their esters and / or salts and / or anhydrides.

[0098] According to one embodiment, the useful composition according to the invention preferably - a conjugate 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 polylysine, polyethylene glycol, polyornithine, polyarginine, or polyhistidine, and - Micelles formed by at least one of these conjugates, including micelles encapsulating farnesyl cysteine and cholesterol.

[0099] Preferably, at least one of the micelles is formed by an amphiphilic conjugate, and each of the amphiphilic conjugates consists of at least one hydrophobic molecule covalently conjugated to a molecule of a polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0100] Thus, according to a particular embodiment of the present invention, at least one micelle is formed by an amphiphilic conjugate, and each of the amphiphilic conjugates consists 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 polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

[0101] In particular, this configuration - increases the in vivo half-life of the active molecule of the composition, - targets the tissue or cell on which the molecules of the composition act, - enables enhancing the stability and bioavailability of the active ingredient.

[0102] Thus, the efficacy of the composition is enhanced, it is possible to administer a lower dose, and reduce the acute or chronic toxicity of the active molecule contained in the composition.

[0103] Preferably, one or more polymers conjugated to the molecule are selected from poly-L-lysine, polyethylene glycol, poly-L-ornithine, poly-L-arginine, and poly-L-histidine.

[0104] The polylysine used in the composition according to the present invention is preferably poly-L-lysine. The polylysine is preferably linear. In particular, the polylysine used can be epsilon-poly-L-lysine, preferably poly-L-lysine having a molecular weight of 12,000 to 20,000 Da. The polylysine used in the composition according to the present invention can be polylysine having bromide, chloride or TFA, trifluoroacetic acid, as a counter ion.

[0105] Thus, according to one variant form, composition C1 comprises at least - A: a conjugate as follows, each conjugate consisting of a molecule covalently bonded to polylysine: - one or more oleyl-poly-L-lysine conjugates, - one or more palmitin-poly-L-lysine conjugates, - one or more lauryl-poly-L-lysine conjugates, - one or more azelail-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: micelles, preferably containing farnesyl cysteine and cholesterol, and / or esters of these molecules, encapsulated in micelles formed by one or more of the conjugates listed in list A.

[0106] In this composition, poly-L-lysine can be replaced by another polylysine, or polyethylene glycol, poly-L-ornithine, poly-L-arginine, or poly-L-histidine.

[0107] The composition for use according to the present invention can be in solid or liquid form. When in liquid form, the composition contains at least water and the above-described components. The solid form is preferably obtained from the liquid form, preferably by lyophilization. When the composition is in liquid form, contains micelles, and is lyophilized into a solid form, the micelles are re-formed when the solid form of the composition is returned to an aqueous solution.

[0108] The composition for use according to the present invention also preferably contains at least one pharmaceutically acceptable excipient. The excipients can be selected in particular to meet the pH and osmotic pressure requirements of solutions for injection into humans or animals. For example, they can be an acid or a base for adjusting the pH, or NaCl for adjusting the osmotic pressure.

[0109] The composition for use according to the present invention, in particular composition C1, is intended to be administered to humans or animals and is thus in a form suitable for such administration. When in liquid form, it is preferably suitable for subcutaneous or intravenous administration, particularly intravenous injection, and is packaged in a suitable container known to those skilled in the art for packaging this type of product. The composition according to the present invention can also be administered in liquid form via a pump such as an insulin pump.

[0110] When in solid form, preferably - preferably in the form of a patch, formulated and packaged to be suitable for transdermal administration, or - suitable for nasal administration in the form of a powder, or - suitable for sublingual administration in the form of a powder or a tablet, or - preferably in the form of a mucoadhesive tablet, formulated and packaged to be suitable for transmucosal absorption administration.

[0111] Manufacturing method The useful composition according to the present invention can be produced by any suitable method.

[0112] When the molecules constituting the composition are used as they are in a solvent, they can all be mixed together in the solvent.

[0113] When some of the molecules in the composition are conjugated to a polymer and the rest are conjugated into micelles, the production method includes the following steps: - a. Step of preparing an amphiphilic premix: in an aqueous solution - One or more butyrate-poly-L-lysine conjugates, - One or more lactate-poly-L-lysine conjugates, - Step of mixing one or more propionate-poly-L-lysine conjugates, - b. Optionally, at least farnesylcysteine and cholesterol are added to the amphiphilic premix and stirred to form micelles formed by one or more of the conjugates of the amphiphilic premix encapsulating farnesylcysteine and cholesterol.

[0114] Preferably, the production method includes the following steps: - a. Step of preparing an amphiphilic premix: 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 selected from the following: - One or more oleyl-poly-L-lysine conjugates - One or more palmitin-poly-L-lysine conjugates - One or more lauryl-poly-L-lysine conjugates - One or more azelail-poly-L-lysine conjugates - one or more palmitoyl - 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 linoleyl - poly - L - lysine conjugates, and - b. adding at least farnesylcysteine and cholesterol to the amphiphilic premix and stirring to form micelles formed by one or more of the conjugates of the amphiphilic premix encapsulating farnesylcysteine and cholesterol, can be included.

[0115] One variant of the present invention lies in utilizing the amphiphilic nature of most of the individual components to create an amphiphilic premix that enables controlled solubilization of hydrophobic species. The solubilization process according to a preferred embodiment consists of the controlled addition of hydrophobic molecules to the amphiphilic premix and allowing the time necessary for their dissolution while stirring.

[0116] Poly - L - lysine can be replaced by another polylysine, or polyethylene glycol, poly - L - ornithine, poly - L - arginine, or poly - L - histidine.

[0117] Preferably, the stirring is carried out for at least 60 minutes, more preferably for 5 - 20 minutes, preferably at a stirring speed of 900 revolutions per minute or less, particularly at a stirring speed of 50 - 800 revolutions per minute.

[0118] According to a preferred embodiment, the manufacturing method according to the present invention also includes step c. of separating the soluble phase from the insoluble phase in order to recover the soluble phase. In this case, the insoluble phase is removed, and the soluble phase constitutes the composition according to the present invention. Specifically, a physical separation process (filtration, ultrafiltration) is preferably carried out to ensure the isolation of the soluble fraction containing both the amphiphilic premix and the solubilizing molecules.

[0119] Subsequently, the composition in liquid form can be lyophilized or dehydrated to a solid form, preferably by slow lyophilization for, for example, 12 to 36 hours.

[0120] Furthermore, if the active molecule-polymer conjugates of the composition are not incorporated into the premix in step a, they can be added to the mixture after step b, that is, after the formation of micelles and the encapsulation of farnesyl cysteine and cholesterol.

[0121] The active molecule-polymer conjugates can be produced by any means known to those skilled in the art for covalently bonding the molecules to the polymer according to their chemical properties. 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) via an amide bond.

[0122] Exemplary embodiments of amide, urea, and carbamate bonds are described, for example, as follows. - 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 bond), - Zhuxian Z, Jianbin T, Qihang S, William J. A multifunctional PEG-PLL drug conjugate forming redox-responsive nanoparticles for intracellular drug delivery, Issue 38, 2015. Journal of Materials Chemistry B (amide bond), - Scheper V, Wolf M, Scholl M, Kadlecova Z, Perrier T, Klok HA, Saulnier P, Lenarz T, Stover T. Potential novel drug carriers for inner ear treatment: hyperbranched poly-lysine and lipid nanocapsules. Nanomedicine (Lond). 2009 Aug;4(6):623-35 (urea bonds), - Stephanie Gac-Breton, Jean Coudane, Mahfoud Boustta & Michel Vert (2004) Norfloxacin-Poly(l-Lysine Citramide Imide) Conjugates and Structure-dependence of the Drug Release, Journal of Drug Targeting, 12:5, 297-307, (carbamate bond), - Elmore, W.M. (2013). Nanoparticles Stabilized with MPEG-Polylysine Carbamate: Synthesis and Characterization, (carbamate bond), - Ning-Ping Huang, Janos Voros, 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 bond).

Example

[0123] Examples of compositions suitable for use according to the present invention are as follows.

[0124] Example 1 Preferred exemplary compositions according to the present invention are shown below.

[0125]

Table 1

[0126] 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 stirred for 10 - 20 minutes depending on the solubility of the conjugate.

[0127] Example 2 Preferred exemplary compositions according to the present invention are shown below.

[0128]

Table 2

[0129] 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 stirred for 10 - 20 minutes depending on the solubility of the conjugate.

[0130] Example 3 Preferred exemplary compositions according to the present invention are shown below.

[0131] [Table 3]

[0132] 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 stirred for 10 - 20 minutes according to the solubility of the conjugate.

[0133] Example 4 Exemplary compositions suitable for testing in mice are shown below.

[0134] [Table 4]

[0135] 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 stirred for 10 - 20 minutes according to the solubility of the conjugate.

[0136] Example 5 Preferred exemplary compositions according to the present invention are shown below.

[0137] [Table 5]

[0138] 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 stirred for 10 - 20 minutes according to the solubility of the conjugate.

[0139] Example 6: Exemplary Composition C1 Useful exemplary Composition C1 according to the present invention is shown below.

[0140] To prepare a stock of 300 mL of Composition C1, the following amounts of the conjugate were weighed. All conjugates were weighed using an analytical balance under a laminar flow hood.

[0141]

Table 6

[0142] Each conjugate was weighed into a sterile 50 mL centrifuge tube and dissolved in 30 mL of water using a vortex mixer. Each conjugate was stirred for 10 - 20 minutes depending on the solubility of the conjugate.

[0143] In parallel, cholesterol and farnesyl cysteine molecules were separately dissolved in absolute ethanol at a concentration of approximately 50 mg / mL and filtered through a 0.22 μm filter. The solvent was evaporated using a rotary evaporator. As shown in Table 3, the required amount of dry solid was weighed under a laminar flow hood.

[0144]

Table 7

[0145] To a sterile 500 mL flask equipped with a magnetic bar, 23.8 mL of each pre - prepared conjugate solution was added while magnetic stirring (700 revolutions per minute). The corresponding amount of (pre - freeze - dried) salt in 300 of PBS was added to the conjugate solution while magnetic stirring (700 revolutions per minute). Cholesterol and farnesyl cysteine were added and the mixture was stirred for 25 hours (700 revolutions per minute).

[0146] 30 mL of Formulation C1 10x was collected under a laminar flow hood and added to a new sterile 500 mL flask.

[0147] 270 mL of sterile PBS was added to obtain Formulation C1 1x. The formulation was stirred for 30 minutes.

[0148] Example 7: Exemplary Composition C1 An exemplary composition C1 suitable for testing in mice is shown below.

[0149] To prepare a 300 mL stock of composition C1, the following amounts of conjugate were weighed. All conjugates were weighed using an analytical balance under a laminar flow hood.

[0150]

Table 8

[0151] Each conjugate was weighed into a sterile 50 mL centrifuge tube and dissolved in 30 mL of water using a vortex mixer. Each conjugate was stirred for 10 - 20 minutes depending on the solubility of the conjugate.

[0152] In parallel, cholesterol and farnesyl cysteine molecules were separately dissolved in absolute ethanol at a concentration of approximately 50 mg / mL and filtered through a 0.22 μm filter. The solvent was evaporated using a rotary evaporator. The required amount of dry solid was weighed under a laminar flow hood.

[0153]

Table 9

[0154] To a sterile 500 mL flask equipped with a magnetic bar, 23.8 mL of each pre - prepared conjugate solution was added while magnetically stirring (700 revolutions per minute). The corresponding amount of (pre - lyophilized) salt in 300 of PBS was added to the conjugate solution while magnetically stirring (700 revolutions per minute).

[0155] Cholesterol and farnesyl cysteine were added and the mixture was stirred for 25 hours (700 revolutions per minute).

[0156] 30 mL of formulation C1 10x was collected under a laminar flow hood and added to a new sterile 500 mL flask.

[0157] 270 mL of sterile PBS was added to obtain Formulation C1 1x. The formulation was stirred for 30 minutes.

[0158] The flasks were prepared according to the following plan using a sterile calibrated 5 mL pipette. C1 1X = 83 flasks, 2 mL each. C1 10X = 83 flasks, 2 mL each.

[0159] 166 flasks were placed in a steel freeze-drying unit and a 1-day freeze-drying cycle was initiated.

[0160] Example 8: Molecular analysis of mouse fecal microbiota (wild-type - WT, SOD1, SOD1 + the composition according to the present invention).

[0161] SOD1 mice are an animal model of axonal degeneration consisting of transgenic mice expressing a mutant form of the human gene for superoxide dismutase. This is an in vivo reference model for studying amyotrophic lateral sclerosis.

[0162] In this test, Compositions C1 (described in Tables 8 and 9) and C2 (described in Table 10) were injected sequentially at 1-hour intervals starting with Composition C1.

[0163]

Table 10

[0164] Feces from 5 male and 5 female mice in each group (WT, SOD1, SOD1 + C1 low dose, and SOD1 + C1 high dose) were analyzed at T0 (= T6), T3 weeks (T9), and 10 weeks (T16), i.e., a total of 55 samples.

[0165] In the first step, total genomic DNA was extracted from each fecal sample using the method of Godon (Godon et al., Appl Envion. Microbiol, 1997). The quality of the extracted DNA was evaluated after electrophoresis of the samples through an agarose gel. Subsequently, the DNA concentration was determined using NanoDrop technology.

[0166] Subsequently, the obtained DNA samples were 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).

[0167] The taxonomic assignment of each obtained bacterial sequence (or OTU) was performed using the silva_nr99_vl38.1 (https: / / www.arb-silva.de / ) and GTDB v202 (https: / / gtdb.ecogenomic.org / ) databases.

[0168] The rANOMALY pipeline (Theil S and Rifa E. rANOMALY: AmplicoN wOrkflow for Microbial community AnaLYsis [version 1; peer review: 2 approved] m F1000Research 2021, 10:7 https: / doi.org / 10.12688 / f1000research.27268.1) based on DADA2 was used to process all of the sequences, whether to estimate the abundance of bacterial phyla / families or to perform statistical analyses (composition, diversity analysis, differential abundance analysis).

[0169] The comparison of bacterial diversity and richness between the microbiota obtained from treated and untreated mice was performed by calculating the Shannon index. Two multivariate analysis methods (ANOVA and non-metric multidimensional scaling = MDS method) and PLS-DA (partial least squares discriminant analysis) from the mixOmics package were used to reveal the structural differences (beta diversity) between the populations.

[0170] The analysis of the differential abundances of OTUs was performed using normalization and estimation tools from the DESeq2 package [Love, M. I., Huber, W., Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 Genome Biology 15(12):550; 2014],

[0171] Microbiota - Clinical Parameters The potential correlations between each of the measured clinical and physiological parameters and the molecular composition of the microbiota were investigated for each of the female mice (5 mice per group) in which the microbiota was analyzed.

[0172] Several analytical methods were tested based on rank correlation (Spearman's rank correlation).

[0173] The method employed was the regularized canonical correlation analysis (rCCA) provided by the mixOmics package (Rohart F, Gautier B, Singh A, and Le Cao K - A, 2017: mixOmics: An R package for ’omics feature selection and multiple data integration. PLoS computational biology 13(11):e1005752). This method enables cross - validation of the analysis results for the correlations between the abundances of OTUs and the various measured clinical parameters. The resulting findings are presented in the form of a heatmap indicating significant positive or negative correlations between the abundances of OTUs and the clinical parameters.

[0174] Results: Comparison of the composition of the fecal microbiota of "SOD1" KO mice and the composition of the fecal microbiota of "wild - type" (WT) mice Analysis of the relative abundances of the phyla, orders, families, or genera of bacteria present in SOD1 mice compared to WT mice indicates that the composition of the SOD1 microbiota is different from that of WT mice. - SOD1 has fewer Firmicutes and more Bacteroidetes than WT. Among the Bacteroidetes, SOD1 mice harbor more Bacteroidetes than WT mice, particularly the Muribaculaceae family, and many Porphyromonadaceae families including Muribaculum. - Among the Firmicutes, the relative abundances of the Lachnospiraceae family (Shaedlerella and Acetifractor) and Erysipelotrichales decrease, while Lactobacillales such as Limolactobacillus and the Oscillospiraceae family, as well as the Eggerthellaceae family, show increased abundances compared to WT mice.

[0175] Analysis of the diversity (alpha diversity) and richness (beta diversity) of the fecal microbiota of SOD1 mice compared to WT mice made it possible to determine the following. - There is no significant difference in bacterial diversity (indices: Chao1, observed, Shannon index, and Inv Simpson index) between SOD1 and WT (alpha diversity). - There are significant differences in the richness of bacterial species (Jaccard: p = 0.01 and Bray-Curtis: p = 0.01) (beta diversity). The time effect (T0, T6, and T16) was not demonstrated.

[0176] The Jaccard and Bray-Curtis indices or distances are two well-known metrics for measuring the similarity, dissimilarity, and diversity among multiple samples.

[0177] Finally, PLS-DA analysis showed that the fecal microbiota of WT mice and SOD1 mice are different (Figure 1).

[0178] In conclusion, compared with WT mice, SOD1 mice have a disrupted gut microbiota, with less bacterial richness, a higher proportion of Gram-negative bacteria (Muribaculaceae and Muribaculum), and Gram-positive bacteria from the Lactobacillales and Eggerthellaceae, but a lower proportion of the Ruminococcaceae and Erysipelotrichaceae.

[0179] Results: Effect of administration of composition C1 (in combination with C2) at high or low doses in SOD1 mice A - Administration of 10× high dose of C1 (in combination with C2) Compositions C1 and C2 are described in Tables 7 - 10.

[0180] Administration of the high dose of composition C1 (in combination with C2) for 3 weeks resulted in the following.

[0181] - An increase in the relative abundance of the following: * Bacteroidales, particularly Muribaculaceae * Oscillospiraceae * COE1 - A decrease in the abundance of the following: * Lactobacillaceae (Lactobacillus genus) * Ruminococcaceae

[0182] Administration of the high dose of composition C1 for 3 weeks appears to enhance the disruption observed in SOD1 mice.

[0183] B - Administration of 1× low dose of C1 (in combination with C2) Administration of the low dose of C1 (in combination with C2) to SOD1 mice also caused changes in the relative abundance of specific microbiota. Thus, the following were observed. - An increase in the relative abundance of the following: * Ruminococcaceae * Oscillospiraceae * GAC-485 - Decrease in the following relative abundances: * Order Lactobacillales

[0184] The low dose appears to partially rebalance the gut microbiota of SOD1 mice to approach the composition of the microbiota of wild-type (WT) mice, particularly by causing an increase in Ruminococcaceae and a decrease in Lactobacillales. This effect appears to be progressive as it is more pronounced 10 weeks after treatment than 3 weeks after treatment.

[0185] Results: Comparison of bacterial diversity and richness in treated and untreated SOD1 mice. Analysis of the alpha diversity of fecal microbiota did not demonstrate a significant effect from administration of C1 (in combination with C2) at low and high doses on bacterial diversity in SOD1 mice. A significant difference (Shannon index, p = 0.02) was observed only between the fecal microbiota of mice treated with the low dose and mice treated with the high dose of C1 (in combination with C2) (Figures 2 and 3).

[0186] Analysis of beta diversity enabled demonstration of a significant difference between untreated SOD1 mice and treated SOD1 mice. - Control vs. high dose of C1 (in combination with C2) (p = 0.02 Jaccard and p = 0.01 Bray-Curtis) - Low dose of C1 (in combination with C2) vs. high dose of C1 (in combination with C2) (p = 0.02 Jaccard and p = 0.01 Bray-Curtis) - There was no significant difference between the untreated control and the low dose of C1 (in combination with C2).

[0187] Results: Comparison between wild-type mice and SOD1 mice treated or untreated with different doses of C1 (in combination with C2) PLS-DA of the compositions of all the microbiota studied made it possible to show that these microbiota can be divided into four different groups, and the microbiota of mice treated with low doses of C1 (in combination with C2) is approaching (being within the same cluster as) the microbiota of wild-type mice (Figure 4).

[0188] Conclusion This study made it possible to show that the composition of the microbiota of SOD1 mice is different from that of wild-type mice and that these changes affect the richness rather than the diversity of the microbiota. The microbiota of SOD1 mice appears to be disrupted as the relative abundances of certain major bacterial groups change. This is characterized, in particular, by an increase in the Muribaculaceae and Lactobacillales (Oscillospiraceae) at the expense of the Ruminococcaceae and Erysipelotrichaceae. PLS-DA further demonstrated that the microbiota of wild-type and SOD1 mice are different.

[0189] The three-week administration of high doses of C1 (in combination with C2) to SOD1 mice seems to amplify the differences observed between wild-type and SOD1 mice (increase in Muribaculaceae and Oscillospiraceae and decrease in Ruminococcaceae). The composition of the microbiota of SOD1 mice treated with high doses of C1 (in combination with C2) is very clearly different from that of SOD1 controls as well as that of wild-type mice, and mice in this group treated with high doses are substantially separated from the others (the most disrupted microbiota).

[0190] The administration of low doses of C1 (in combination with C2) to SOD1 mice over 3 to 10 weeks seems to partially restore the disrupted microbiota balance observed in SOD1 control mice. Specifically, an increase in Ruminococcaceae and a decrease in the Lactobacillus microbiota are observed in SOD1 mice compared to wild-type mice. Thus, the composition of the microbiota of SOD1 mice treated with low doses of C1 (in combination with C2) becomes closer to that of wild-type mice, and the microbiota of these two groups are classified into the same cluster.

[0191] These preliminary results are interesting regarding the administration of the lowest dose in SOD1 model mice. Specifically, at this dose, C1 (in combination with C2) appears to have a beneficial effect on the disrupted microbiota of SOD1 mice.

[0192] Example 9: Study on the correlation between symptoms and the molecular composition of the microbiota in SOD1 mice after administration of low-dose C1 (in combination with C2) The purpose of the study was to analyze the potential correlation between the molecular composition of the gut microbiota and clinical parameters measured in SOD1 mice versus wild-type mice and SOD1 mice treated or untreated with low-dose C1 (in combination with C2) for 10 weeks.

[0193] This study was conducted in female mice whose fecal microbiota had been previously molecularly analyzed (5 females were sampled from each cage from each group (WT, SOD1, SOD1-low dose) at T0 (=T6), T3 weeks (T9), and 10 weeks (T16)).

[0194] The tests were performed at least 1 hour after the first daily dose of the test compound or vehicle compound.

[0195] One-day sessions included a 5-minute practice run at 4 rpm on a rotarod apparatus (AccuScan Instruments, Columbus, USA). One hour later, the animals were tested for three consecutive acceleration trials at speeds from 0 to 40 rpm for 6 minutes over 360 seconds, with a trial interval of at least 30 minutes. The latency to fall off the rod was recorded.

[0196] Study on the correlation in SOD1 mice and wild-type mice. Regular Canonical Correlation Analysis (RCCA) mainly enabled the demonstration of a significant negative correlation between the clinical score, movement distance, and recovery time after the rotarod test and the Desulfovibrionaceae family, as well as a smaller negative correlation with the Prevotellaceae family, in WT mice and SOD1 mice. Therefore, the higher the abundance of these bacterial families, the lower the clinical score, and the same applies to the movement distance and recovery time (Figure 5).

[0197] The relative abundance of the Desulfovibrionaceae family (and the Prevotellaceae family) was low compared to other bacterial families, but the presence of OTUs of this family could only be found in SOD1 mice (not detected in wild-type mice). This may suggest that the Desulfovibrionaceae family plays a potential role in the pathophysiology of SOD1 mice.

[0198] Study on the correlation in SOD1 mice treated with low-dose C1 (in combination with C2) for 10 weeks or untreated RCCA performed in SOD1 control mice and mice treated with low-dose PL enabled the demonstration of a strong positive correlation between the Ruminococcaceae family and the movement distance of the mice. The richer this family is, the longer the movement distance of the mice. Considering that one of the main effects of the composition according to the present invention on the gut microbiota of SOD1 mice is to increase the abundance of Ruminococcaceae species, the observed correlation suggests that this bacterial family plays a role in the recovery of the ability of SOD1 mice to move a distance close to that of wild-type mice (Figure 6).

[0199] Demonstration of the negative correlation between the Desulfovibrionaceae family and the severity of symptoms in SOD1 mice suggests that species from this sulfide-producing family may be involved in pathophysiology, as observed in other pathologies such as irritable bowel syndrome or chronic inflammatory bowel diseases (Crohn's disease, ulcerative colitis). In parallel, the lower abundance of species from the Ruminococcaceae family in SOD1 mice may lead to a decrease in the concentration of colonic butyrate, since most species from the Ruminococcaceae family produce this metabolite with demonstrated effects on health.

[0200] Administration of the composition according to the invention at a low dose over 10 weeks is accompanied by changes in the composition of the microbiota of SOD1 mice, in particular an increase in the abundance of the Ruminococcaceae family and the disappearance of the Desulfovibrionaceae family. This correlates with a significant improvement in the ability of the mice to move the same distance as wild-type mice. Thus, administration of the composition according to the invention partially restores the balance of the altered microbiota of SOD1 mice, brings about metabolic changes, and reduces the severity of specific symptoms in SOD1 mice or restores them to a normal phenotype.

Claims

**Claim 1** A composition comprising at least - lactic acid, and / or a salt and / or ester and / or anhydride of lactic acid, - butyric acid, and / or a salt and / or ester and / or anhydride of butyric acid, - propionic acid, and / or a salt and / or ester and / or anhydride of propionic acid, for use in the prevention and / or treatment of dysbiosis of the gut microbiota in humans and animals. **Claim 2** The composition for use according to claim 1, wherein the prevention and / or treatment is characterized by an increase in the proportion of butyrate-producing bacteria and / or a decrease in the proportion of sulfide-producing bacteria in the gut microbiota. **Claim 3** The composition for use according to claim 1 or 2, wherein the dysbiosis of the gut microbiota is characterized by excessive sulfide-producing bacteria and a lack of butyrate-producing bacteria. **Claim 4** The composition for use according to any one of claims 1 to 3, wherein the dysbiosis of the gut microbiota is also characterized by an increase in gut inflammation. **Claim 5** The composition for use according to any one of claims 1 to 4 for the prevention and / or treatment of at least one neurodegenerative disease and / or gut diseases associated with dysbiosis of the gut microbiota. **Claim 6** The composition for use according to claim 5, wherein the neurodegenerative disease is selected from Lou Gehrig's disease, amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease. **Claim 7** The composition for use according to claim 5, wherein the gut disease is selected from Crohn's disease, chronic inflammatory bowel disease, hemorrhagic proctocolitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis, and gluten intolerance. **Claim 8** The composition for use according to any one of claims 1 to 7 as a drug or dietary supplement in humans or animals. **Claim 9** The composition for use according to any one of claims 2 to 8, wherein the sulfide-producing bacteria belong to the family Desulfovivrionaceae and the butyrate-producing bacteria belong to the family Lachnospiraceae. **Claim 10** The composition for use according to claim 8 or 9, wherein the bacteria belonging to the family Desulfovibrionaceae are less than 0.01% of the total bacteria present in the intestinal microbiota.

11. The composition for use according to any one of claims 1 to 10, comprising at least one polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

12. The composition for use according to any one of claims 1 to 11, wherein at least one molecule 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 polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine.

13. The composition is - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesylcysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid, - acetic acid, and combinations thereof, and further comprises at least one molecule selected from The composition according to any one of claims 1 to 12, wherein the molecule(s) is / are potentially in the form of one or more salts and / or esters and / or anhydrides of these molecules.

14. The composition comprises at least the following molecules: - oleic acid, - palmitic acid, - lauric acid, - linoleic acid, - azelaic acid, - farnesylcysteine, - palmitoleic acid, - cholesterol, - thioctic acid, - myristic acid, - orotic acid, - pyruvic acid, - acetic acid, and combinations thereof, and / or comprises one or more salts and / or esters and / or anhydrides of these molecules, and is the composition according to any one of claims 1 to 13.

15. At least one molecule of a 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 polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine. A composition for use according to any one of claims 1 to 14, characterized in that.

16. A composition according to any one of claims 12 to 15, characterized in that it contains micelles encapsulating at least farnesyl cysteine and / or cholesterol and / or esters of these molecules.

17. At least one of the micelles is formed by an amphiphilic conjugate, and each of the amphiphilic conjugates consists of at least one hydrophobic molecule covalently conjugated to a molecule of a polymer selected from polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine. A composition according to any one of claims 1 to 16, characterized in that.

18. At least one micelle is formed by an amphiphilic conjugate, and each of the amphiphilic conjugates consists 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 polylysine, polyethylene glycol, polyornithine, polyarginine, and polyhistidine. A composition according to any one of claims 1 to 17, characterized in that.

19. A composition for use according to any one of claims 1 to 18, characterized in that at least one molecule is covalently conjugated to at least one polymer.

20. The composition is at least - A. The following conjugates, each conjugate consisting of a molecule covalently bonded to polylysine: - One or more oleyl-poly-L-lysine conjugates, - One or more palmitin-poly-L-lysine conjugates, - one or more lauryl - poly - L - lysine conjugates, - one or more azelail - 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. - farnesylcysteine and cholesterol encapsulated in micelles, and / or esters of these molecules, characterized in that it comprises a composition for use according to any one of claims 1 to 19.

21. A composition for use according to any one of claims 1 to 20, characterized in that farnesylcysteine and cholesterol, and / or esters of these molecules are encapsulated in micelles formed by one or more of the conjugates from List A.

22. A composition for use according to claim 20 or 21, characterized in that the poly - L - lysine is replaced by another polylysine or by polyethylene glycol, poly - L - ornithine, poly - L - arginine, or poly - L - histidine.

23. A composition for use according to any one of claims 1 to 22, characterized in that the composition comprises at least one pharmaceutically acceptable excipient.

24. A composition for use according to any one of claims 1 to 23, characterized in that the composition is in liquid form or solid form.

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