Composition for preventing and / or treating a pathological dysbiosis of the intestinal microbiota

Abstract

To meet the needs of new therapies for preventing or treating pathological dysbiosis of the intestinal microbiota, in particular for treating or preventing neurodegenerative or intestinal diseases. The invention relates to the use of compositions comprising a plurality of specific molecules for treating a pathological dysbiosis of the intestinal microbiota. The pathological dysbiosis of the intestinal microbiota can be characterized by an excess of sulfide-producing bacteria and a deficit in butyrate-producing bacteria.

Description

[0001] COMPOSITION FOR THE PREVENTION AND / OR TREATMENT OF PATHOLOGICAL DYSBIOSIS OF THE INTESTINAL MICROBIOTA

[0002] Technical field

[0003] The invention relates to the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota. In particular, the invention relates to the use of compositions comprising several specific molecules for treating pathological dysbiosis of the intestinal microbiota.

[0004] Prior art

[0005] The microbiota is the set of microorganisms (bacteria, archaea; viruses and eukaryotes) which develop in a specific environment.

[0006] The human body contains a multitude of microbiota, particularly in the skin, mouth, vaginal cavity, throat and digestive system.

[0007] 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.

[0008] The intestinal microbiota is the microbiota that houses a large part of the 10,000 billion microorganisms populating the human body.

[0009] 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 intestinal microbiota. The characterization of all the microbial genomes found in the intestine has led to the discovery of more than a thousand different species, the vast majority of which are bacterial species.

[0010] In-depth analysis of the intestinal microbiota has demonstrated that, like a fingerprint, the intestinal microbiota is specific to each individual and depends on their environment as well as their genetic heritage.

[0011] Once established, the composition of the microbiota remains relatively stable throughout an individual's lifetime. Variations are generally less than 5% of the total microbiota.

[0012] Thus, pathogenic states of the intestinal microbiota are difficult to detect because they come from these particular species.

[0013] When a patient presents with a disease, we can find an imbalance in the pathogenic intestinal flora called pathological dysbiosis; the threshold of imbalance is difficult to establish, as are the variations of genera and species unless there is a biomarker of the disease. Thanks to the removal of technical obstacles, 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 appearance of numerous diseases.

[0014] Indeed, the intestinal microbiota plays an essential role in the functioning of the body. The microorganisms that compose it have a wide range of actions, from the breakdown of food to epithelial repair and the regulation of metabolism.

[0015] However, 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.

[0016] The central nervous system-gut interaction is a major component in the onset of certain pathologies. In Parkinson's disease, the link between pathological dysbiosis of the intestinal microbiota and the onset of motor disorders as well as neuroinflammation has been demonstrated.

[0017] Pathological dysbiosis of the intestinal microbiota has been demonstrated in other neurodegenerative diseases, notably in patients with amyotrophic lateral sclerosis.

[0018] Furthermore, it is now known that intestinal diseases and more particularly inflammatory bowel diseases, Crohn's disease and food intolerance, particularly gluten intolerance, are linked to an intestinal microbiota that is very depleted in microorganisms and therefore the presence of pathological dysbiosis.

[0019] Currently, the solutions proposed to prevent or combat dysbiosis essentially consist of administering probiotics and / or prebiotics, particularly to treat intestinal diseases, but their effectiveness is very variable and depends on the patient's basic intestinal flora.

[0020] Another promising solution to combat pathological dysbiosis is fecal transplantation. However, the effect on the intestinal microbiota requires at least two transplants per week to achieve a lasting effect.

[0021] Furthermore, with regard to degenerative diseases specifically, these are often very burdensome for the patient. Some symptoms can be reduced, but these diseases are never completely cured and there is currently no satisfactory treatment. Many chemical agents have therapeutic properties but either their toxicity, their lifespan, or their rapid elimination make them unsuitable for the treatment of degenerative diseases. Indeed, while they are multifactorial, current treatments only target one of the factors causing these diseases, and do not seek to act in a more comprehensive manner with a systemic approach directly targeting the causes of the disease.For example, Lou Gehrig's disease or amyotrophic lateral sclerosis is linked to many factors, in particular to oxidative stress, mitochondrial dysfunction, neuroinflammation, excitotoxicity, oligodendrocyte dysfunction and degeneration, impaired proteostasis, an impaired DNA repair system, alterations in nucleocytoplasmic RNA and in RNA bound to transport proteins, defective axonal transport, defective vesicular transport and pathological dysbiosis of the intestinal microbiota. However, currently, the recommended treatment for Lou Gehrig's disease is Riluzole® which acts only on the inhibition of glutamate release to combat excitotoxicity, and in addition no action on the intestinal microbiota is envisaged. The observation is identical for other degenerative diseases.

[0022] There is therefore a significant need for a solution to prevent or treat pathological dysbiosis of the intestinal microbiota, particularly to treat or prevent neurodegenerative or intestinal diseases, to meet an essential demand for new therapies.

[0023] The objective of the invention is therefore to meet all of these needs and to overcome the drawbacks and limitations of the prior art.

[0024] Summary of the invention

[0025] 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 suitable for uses for treating or preventing diseases associated with pathological dysbiosis of the intestinal microbiota, in particular neurodegenerative or intestinal diseases.

[0026] For this purpose, the invention relates to a composition comprising at least:

[0027] - lactic acid, and / or a salt and / or an ester and / or an anhydride of lactic acid,

[0028] - butyric acid, and / or a salt and / or an ester and / or an anhydride of butyric acid, and

[0029] - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid, for its use in humans or animals in the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0030] Advantageously, the composition according to the invention is capable of restoring the balance of the microbiota of an individual presenting a pathological dysbiosis of the intestinal microbiota. 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 sulfide-producing bacteria and a deficit of butyrate-producing bacteria.

[0031] Said treatment according to the invention is thus preferentially characterized in the intestinal microbiota by an increase in the proportion of bacteria producing butyrate and a decrease in the proportion of bacteria producing sulphide.

[0032] Advantageously, the effect on butyrate-producing bacteria and sulfide-producing bacteria provides a synergistic action to treat pathological dysbiosis and in particular pathological dysbiosis associated with pathologies.

[0033] More particularly, the composition can be used in humans or animals in the prevention and / or treatment of at least one neurodegenerative disease and / or an intestinal disease associated with a pathological dysbiosis of the intestinal microbiota. According to a particularly suitable variant, the composition can be used in humans or animals in the prevention and / or treatment of at least one neurodegenerative disease and / or an intestinal disease associated with a pathological dysbiosis of the intestinal microbiota in which the human or animal has an intestinal dysbiosis characterized in the intestinal microbiota by an increase in the proportion of bacteria producing sulfide and a decrease in the proportion of bacteria producing butyrate.

[0034] 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.

[0035] The composition according to the invention can thus be used as a medicine or food supplement in humans or animals.

[0036] 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.

[0037] 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. 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, can also comprise at least one molecule chosen from:

[0038] - oleic acid,

[0039] - palmitic acid,

[0040] - lauric acid,

[0041] - linoleic acid,

[0042] - azelaic acid,

[0043] - farnesyl cysteine,

[0044] - palmitoleic acid,

[0045] - cholesterol,

[0046] - thioctic acid,

[0047] - myristic acid,

[0048] - orotic acid,

[0049] - acetic acid, and their combination, said molecule(s) possibly being in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0050] Preferably, the composition for its use comprises the following molecules:

[0051] - oleic acid,

[0052] - palmitic acid,

[0053] - lauric acid,

[0054] - linoleic acid,

[0055] - azelaic acid,

[0056] - farnesyl cysteine,

[0057] - palmitoleic acid,

[0058] - cholesterol,

[0059] - thioctic acid, - myristic acid,

[0060] - orotic acid,

[0061] - acetic acid,

[0062] - butyric acid,

[0063] -lactic acid,

[0064] - propionic acid and / or a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0065] Advantageously, these molecules each act on different factors, allowing for a multifactorial action on the intestinal microbiota. These molecules will thus help to better balance excess or deficient bacterial populations, in particular by reducing intestinal inflammation.

[0066] Advantageously, the molecules of the composition for its use according to the invention can be conjugated to specific polymers, such as poly-lysine, thus making it possible to increase the effectiveness and bioavailability of these molecules.

[0067] Thus, according to a particularly preferred embodiment, the composition for its use according to the invention comprises at least the following conjugates, each conjugate being constituted by a molecule covalently linked to a poly-lysine:

[0068] - one or more Butyrate-Poly-L-Lysine conjugates

[0069] - one or more Lactate-Poly-L-Lysine conjugates

[0070] - one or more Propionate-Poly-L-Lysine conjugates.

[0071] According to one variant, the composition for its use according to the invention comprises at least:

[0072] - A. the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine:

[0073] - one or more Oleyl-Poly-L-Lysine conjugates

[0074] - one or more Palmitic-Poly-L-Lysine conjugates

[0075] - one or more Lauryl-Poly-L-Lysine conjugates

[0076] - one or more Azelayl-Poly-L-Lysine conjugates

[0077] - one or more Palmitoleyl-Poly-L-Lysine conjugates

[0078] - one or more Thioctyl-Poly-L-Lysine conjugates

[0079] - one or more Myristyl-Poly-L-Lysine conjugates - one or more Orotyl-Poly-L-Lysine conjugates

[0080] - one or more Acetate-Poly-L-Lysine conjugates

[0081] - one or more Butyrate-Poly-L-Lysine conjugates

[0082] - one or more Lactate-Poly-L-Lysine conjugates

[0083] - one or more Propionate-Poly-L-Lysine conjugates

[0084] - one or more Linoleyl-Poly-L-Lysine conjugates, and

[0085] - 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.

[0086] The invention also relates to a method for manufacturing these compositions.

[0087] Other features and advantages will emerge from the detailed description of the invention and the examples which follow.

[0088] Brief description of the figures

[0089] [Fig.l] Figure 1 is a graphical representation of a PLS-DA analysis of the comparison of the microbiota of WT and SOD1 mice

[0090] [Fig 2] Figure 2 is a graphical representation of a comparative analysis of the beta diversity of the mouse intestinal microbiota via the MDS - Jaccard method as a function of treatment.

[0091] [Fig 3] Figure 3 is a graphical representation of a comparative analysis of the beta diversity of the mouse intestinal microbiota via the MDS - Bray Curtis method as a function of treatment.

[0092] [Fig 4] Figure 4 is a graphical representation of a comparative analysis of mouse gut microbiota as a function of treatment via PLS-DA analysis.

[0093] [Fig 5] Figure 5 is a graphical representation of a correlation analysis between WT or SOD1 mice based on bacterial families.

[0094] [Fig 6] Figure 6 is a graphical representation of a correlation analysis between low-dose treated and untreated SOD1 mice

[0095] Detailed description of the invention

[0096] Definitions

[0097] For the purposes of the invention, the term "Animal" means any Animal with the exception of the human being (Man). This particularly concerns mammals. For the purposes of the invention, the term "amphiphilic conjugate" means a conjugate formed by a hydrophobic molecule X and a hydrophilic polymer Y, the conjugate thus having an amphiphilic characteristic.

[0098] For the purposes of the invention, the term “molecule X conjugated to a polymer Y” means a molecule X linked by a covalent bond to a polymer Y, said bond being preferably able to be an amide, urea or carbamate bond depending on the chemical nature of the molecule X and the polymer Y.

[0099] By "medical nutrition composition" or "medical nutrition product" or "medical food" or foodstuffs intended for special medical purposes (DADFMS) or Dietary foodstuffs intended for special medical purposes (ADDFMS), within the meaning of the invention, is meant a food intended for therapeutic prevention and / or treatment, used alone or in combination with other therapies. It is a nutritional compound, responding to a particular clinical situation, capable of constituting the exclusive or partial diet of the patients for whom it is intended.

[0100] For the purposes of the invention, “pathological dysbiosis” means a pathological condition characterized by an imbalance in the distribution of bacteria in the intestinal microbiota at the level of phyla, classes, orders, families, genera or species, leading to a risk of other diseases of metabolic origin.

[0101] For the purposes of the invention, "medicine" means a product that has obtained marketing authorization as a product for the prevention and / or treatment of a disease.

[0102] For the purposes of the invention, “microbiota” means the intestinal microbiota.

[0103] For the purposes of the invention, the term "butyrate-producing bacteria" means bacteria capable of producing butyrate by the anaerobic fermentation of non-digestible dietary fibers. Butyrate-producing bacteria mainly belong to the Firmicutes phylum.

[0104] For the purposes of the invention, “sulfide-producing bacteria” means bacteria capable of reducing sulfates to sulfides.

[0105] For the purposes of the invention, “alpha diversity” means the measurement of the number of species present in a given sample.

[0106] For the purposes of the invention, “beta diversity” means the measurement of the diversity of species in a given sample.

[0107] For the purposes of the invention, the term "excess of sulfide-producing bacteria" means a proportion of sulfide-producing bacteria higher than normal, preferably higher than the proportion of sulfide-producing bacteria in an individual not presenting pathological dysbiosis.

[0108] For the purposes of the invention, the term "deficiency in butyrate-producing bacteria" means a proportion of butyrate-producing bacteria lower than normal, preferably lower than the proportion of butyrate-producing bacteria in an individual not presenting pathological dysbiosis.

[0109] Composition for its use

[0110] The invention therefore relates to a composition comprising at least:

[0111] - lactic acid, and / or a salt and / or an ester and / or an anhydride of lactic acid,

[0112] - butyric acid, and / or a salt and / or an ester and / or an anhydride of butyric acid, and

[0113] - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid, for use in humans or animals for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0114] Preferably, the composition is particularly suitable for use in humans for the prevention and / or treatment of pathological dysbiosis of the intestinal microbiota.

[0115] Advantageously, the combination of these three molecules ensures a significant preventive and / or curative effect on pathological dysbiosis of the intestinal microbiota. Lactate, butyrate and propionate are the main short-chain fatty acids resulting from the degradation of fibers by bacteria in the intestinal microbiota. These three fatty acids are involved in numerous cellular functions and are the main energy source for the intestinal epithelium, particularly for the epithelial cells of the intestine and colon. Too low a content of short-chain fatty acids can induce pathological dysbiosis.

[0116] In the context of the invention, pathological dysbiosis of the intestinal microbiota is preferentially characterized by an excess of sulfide-producing bacteria and a deficit of butyrate-producing bacteria.

[0117] The presence of an excess of sulfur-producing bacteria is characteristic of pathological dysbiosis. Indeed, the reduction in the proportion of beneficial microorganisms within the microbiota reveals bacterial populations having a harmful effect or producing harmful metabolites for the body. Thus, patients with pathological dysbiosis have high levels of sulfur-reducing bacteria associated with an increase in the 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.

[0118] The use of a composition according to the invention is thus characterized in the intestinal microbiota by an increase in the proportion of bacteria producing butyrate and a decrease in the proportion of bacteria producing sulfide.

[0119] Advantageously, reducing the proportion of sulfide-producing bacteria helps 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 thus stimulates the microbiota and in particular the bacterial species beneficial to the body. Thus, the simultaneous action on butyrate- and sulfide-producing bacteria allows for a synergistic action on the pathological dysbiosis of the intestinal microbiota. The composition guarantees a long-term effect on the intestinal microbiota by targeting the causes of pathological dysbiosis.

[0120] The composition can be used to prevent or treat any disease that has caused pathological dysbiosis of the intestinal microbiota or that has 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 bacteria producing sulfide and a decrease in the proportion of bacteria producing butyrate, and also preferably characterized by an increase in intestinal inflammation.

[0121] More particularly, the composition comprising at least:

[0122] - lactic acid, and / or a salt and / or an ester and / or an anhydride of lactic acid,

[0123] - butyric acid, and / or a salt and / or an ester and / or an anhydride of butyric acid, and

[0124] - propionic acid, and / or a salt and / or an ester and / or an anhydride of propionic acid can be used in humans or animals in the prevention and / or treatment of at least one disease associated with a pathological dysbiosis of the intestinal microbiota, of at least one disease associated with a pathological dysbiosis of the intestinal microbiota in particular in which the human or animal has an intestinal dysbiosis characterized in the microbiota by an increase in the proportion of bacteria producing sulfide and a decrease in the proportion of bacteria producing butyrate, and in particular of at least one neurodegenerative disease and / or an intestinal disease associated with a pathological dysbiosis of the intestinal microbiota,such as at least one neurodegenerative disease and / or one intestinal disease associated with pathological dysbiosis of the intestinal microbiota in which the Human or Animal presents an intestinal dysbiosis characterized in the microbiota by an increase in the proportion of bacteria producing sulfide and a decrease in the proportion of bacteria producing butyrate.,

[0125] In the context of the invention, neurodegenerative diseases may be chosen from multiple sclerosis, amyotrophic lateral sclerosis also called “Charcot's disease”, Parkinson's disease and Alzheimer's disease.

[0126] Furthermore, intestinal diseases can be chosen from Crohn's disease, chronic inflammatory bowel disease, ulcerative colitis, irritable bowel syndrome, ulcerative colitis, rheumatoid arthritis and food intolerance including gluten intolerance.

[0127] Alternatively, the composition may be used to treat or prevent autism spectrum disorders or spondyloarthritis, particularly in which humans or animals have 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.

[0128] 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.

[0129] The composition thus allows to act 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 allowing to promote the growth of bacteria producing butyrate while restoring membrane permeability.

[0130] The composition can thus be used as a medicine, medical nutrition product or food supplement to rebalance the intestinal microbiota by increasing the proportion of bacteria producing sulphide and a reduction in the proportion of bacteria producing butyrate and or to prevent and combat diseases arising from or caused by this pathological imbalance of the intestinal microbiota.

[0131] In the context of the invention, the sulfide-producing bacteria may be bacteria belonging to the Desulfovibrionaceae family and / or proteolytic bacteria.

[0132] Sulphide-producing bacteria belonging to the Desulfovibrionaceae family may belong to at least one bacterial genus chosen from:

[0133] - Alteridesulfovibrio

[0134] - Aminidesulfovibrio

[0135] - Bilophila - Desulfocurvus

[0136] - Desulfovibrio

[0137] - Frigididesulfovibrio

[0138] - Fundidesulfovibrio

[0139] - Halodesulfovibrio

[0140] - Humidesulfovibrio

[0141] - Lawsonia

[0142] - Maridesulfovibrio

[0143] - Mega lodesulfovi brio

[0144] - Nitratidesulfovibrio

[0145] - Oleidesulfovibrio

[0146] - Paradesulfovibrio

[0147] - Paucidesulfovibrio

[0148] - Pseudodesulfovibrio

[0149] - Solidesulfovibrio.

[0150] Preferably, sulfide-producing bacteria belong to the Desulfovibrionaceae family, the Bilophila genus, and the wadworthia species.

[0151] Thus, the composition according to the invention can be used in humans or animals to reduce the proportion of the bacterial species Bilophila wadworthia in humans or animals presenting intestinal dysbiosis characterized in the microbiota by an increase in the proportion of bacteria producing sulfide.

[0152] The composition also acts on at least one family of butyrate-producing bacteria chosen from:

[0153] - The Lachnospiraeceae

[0154] - The Ruminococcacea

[0155] Butyrate-producing bacteria belonging to the Lachnospiraeceae family may belong to at least one bacterial genus chosen from:

[0156] - Lachnospira

[0157] - Roseburria

[0158] - Eubacterium - Anaerostipes.

[0159] Butyrate-producing bacteria belonging to the Ruminococcacea family may belong to the bacterial genus Faecalibacterium.

[0160] In a particularly preferred embodiment, the composition can be used to increase the proportion of bacteria of the Lachnospiraeceae family and decrease the proportion of bacteria of the Desulfovibriaceae family, preferentially so that the Desulfovibriaceae represent less than 0.01% of the total bacteria present in the intestinal microbiota.

[0161] Advantageously, the composition helps reduce inflammation of the digestive system by restoring sulfide and butyrate concentrations similar to those of a healthy patient.

[0162] 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 chosen from:

[0163] - oleic acid,

[0164] - palmitic acid,

[0165] - lauric acid,

[0166] - linoleic acid,

[0167] - azelaic acid,

[0168] - farnesyl cysteine,

[0169] - palmitoleic acid,

[0170] - cholesterol,

[0171] - thioctic acid,

[0172] - myristic acid,

[0173] - orotic acid,

[0174] - pyruvic acid

[0175] - acetic acid, and their combination, said molecule(s) possibly being in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0176] Preferably, the composition comprises at least 2 molecules, in particular at least 3, even more preferably at least 4 chosen from:

[0177] - oleic acid, - palmitic acid,

[0178] - lauric acid,

[0179] - linoleic acid,

[0180] - azelaic acid,

[0181] - farnesyl cysteine,

[0182] - palmitoleic acid,

[0183] - cholesterol,

[0184] - thioctic acid,

[0185] - myristic acid,

[0186] - orotic acid,

[0187] - pyruvic acid

[0188] - acetic acid, and their combination, said molecule(s) possibly being in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0189] 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.

[0190] For all the molecules present in the compositions for its use according to the invention, when they are mentioned in the present application, they 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.

[0191] According to a particularly preferred embodiment of the invention, the composition comprises at least the following molecules:

[0192] - oleic acid,

[0193] - palmitic acid,

[0194] - lauric acid,

[0195] - linoleic acid,

[0196] - azelaic acid, - farnesyl cysteine,

[0197] - palmitoleic acid,

[0198] - cholesterol,

[0199] - thioctic acid,

[0200] - myristic acid,

[0201] - orotic acid,

[0202] - lactic acid

[0203] - propionic acid

[0204] -butyric acid

[0205] - acetic acid, and their combination, said molecule(s) possibly being in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules.

[0206] 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 decreasing the proportion of bacteria producing sulfide.

[0207] The useful composition according to the invention comprising at least all of the following molecules:

[0208] - oleic acid,

[0209] - palmitic acid,

[0210] - lauric acid,

[0211] - linoleic acid,

[0212] - azelaic acid,

[0213] - farnesyl cysteine,

[0214] - palmitoleic acid,

[0215] - cholesterol,

[0216] - thioctic acid,

[0217] - myristic acid,

[0218] - orotic acid,

[0219] - lactic acid

[0220] - propionic acid - butyric acid

[0221] - acetic acid, and their combination, said molecule(s) being able to be in the form of a salt and / or an ester and / or an anhydride of one or more of these molecules, is hereinafter called composition Cl.

[0222] The composition useful according to the invention, and in particular composition C1, may optionally also comprise pyruvic acid and / or a salt and / or an ester and / or an anhydride.

[0223] Preferably:

[0224] - the quantity of butyric acid in the composition and more particularly in composition Cl is greater than that of each of the other molecules taken individually, and / or

[0225] - the quantity of thioctic acid in the composition and more particularly in composition Cl is greater than that of each of the other molecules taken individually, except butyric acid, and / or

[0226] - the quantity of lauric acid in the composition and more particularly in the composition Cl 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,

[0227] - the quantity of oleic acid on the one hand and lactic acid on the other hand in the composition and more particularly in the composition Cl is greater than that of each of the following molecules taken individually: palmitic acid, lauric acid, linoleic acid, azelaic acid, farnesyl cysteine, palmitoleic acid, cholesterol, myristic acid, orotic acid, acetic acid.

[0228] Preferably, each of the molecules in the composition Cl represents between 0.5 E-05 M and 10 E-05 M.

[0229] According to a particular embodiment of the invention, the composition useful according to the invention, and in particular composition Cl, 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.

[0230] According to a variant, the composition useful according to the invention, and in particular composition C1, is administered with a first so-called “attack” dose of between 7 and 10 mg / mL, preferably between 9 and 10 mg / mL, then a “maintenance” dose of between 0.5 and 5 mg / mL, in particular between 2 and 3 mg / mL. Preferably, the composition useful according to the invention, and in particular C1, may comprise at least one polymer chosen from poly-lysine, poly-ethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0231] More particularly, to improve the solubility, the efficacy and the bioavailability of the molecules of the composition useful according to the invention at least one of the molecules of the composition chosen from butyric acid, lactic acid, propionic acid, the salts of these acids, the esters of these acids and the anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer chosen from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0232] When the composition is composition Cl or a composition comprising the molecules from 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, the salts of these acids, the esters of these acids and the anhydrides of these acids, is covalently conjugated to at least one molecule of a polymer chosen from poly-lysine, poly-ethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0233] 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, the salts of these acids, the esters of these acids and the anhydrides of these acids, are all covalently conjugatable with a polymer chosen from poly-lysine, poly-ethylene glycol, poly-ornithine, poly-arginine and poly-histidine, in particular by an amide, urea or carbamate bond.

[0234] According to a particularly suitable variant, all 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, the salts of these acids and the esters of these acids, when present in the composition useful according to the invention, are covalently conjugated to at least one molecule of a polymer chosen from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0235] The composition useful according to the invention may comprise micelles. When the composition according to the invention and in particular composition C1, comprises farnesyl cysteine ​​and / or cholesterol and / or one of their esters and / or salts and / or anhydrides, the composition useful according to the invention preferably comprises micelles in which at least farnesyl cysteine ​​and / or cholesterol and / or one of their esters and / or salts and / or anhydrides are encapsulated.

[0236] According to one embodiment, the useful composition according to the invention preferably comprises:

[0237] - 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, the salts of these acids, the esters of these acids and the anhydrides of these acids, with poly-lysine, poly-ethylene glycol, poly-ornithine, poly-arginine or poly-histidine, and

[0238] - micelles formed by at least one of these conjugates, said micelles encapsulating farnesyl cysteine ​​and cholesterol.

[0239] 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 chosen from poly-lysine, polyethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0240] 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, the salts, esters and anhydrides of these fatty acids, covalently conjugated to a molecule of a polymer chosen from poly-lysine, poly-ethylene glycol, poly-ornithine, poly-arginine and poly-histidine.

[0241] This configuration allows in particular:

[0242] - to increase the half-life of the active molecules of the composition in the body,

[0243] - to target the tissues or cells on which the composition molecules must act,

[0244] - to increase the stability and bioavailability of the active ingredient.

[0245] 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.

[0246] 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.

[0247] 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 of between 12,000 and 20,000 Da. The poly-lysine used in the compositions according to the invention may be a poly-lysine having a bromide, or chlorine or TFA trifluoroacetic acid counterion.

[0248] Thus, according to one variant, the composition Cl comprises at least:

[0249] -A: the following conjugates, each conjugate consisting of a molecule covalently linked to a poly-lysine:

[0250] - one or more Oleyl-Poly-L-Lysine conjugates

[0251] - one or more Palmitic-Poly-L-Lysine conjugates

[0252] - one or more Lauryl-Poly-L-Lysine conjugates

[0253] - one or more Azelayl-Poly-L-Lysine conjugates

[0254] - one or more Palmitoleyl-Poly-L-Lysine conjugates

[0255] - one or more Thioctyl-Poly-L-Lysine conjugates

[0256] - one or more Myristyl-Poly-L-Lysine conjugates

[0257] - one or more Orotyl-Poly-L-Lysine conjugates

[0258] - one or more Acetate-Poly-L-Lysine conjugates

[0259] - one or more Butyrate-Poly-L-Lysine conjugates

[0260] - one or more Lactate-Poly-L-Lysine conjugates

[0261] - one or more Propionate-Poly-L-Lysine conjugates

[0262] - one or more Linoleyl-Poly-L-Lysine conjugates,

[0263] - 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.

[0264] In this composition, poly-L-Lysine may be replaced by another poly-lysine or by polyethylene glycol, poly-L-ornithine, poly-L-arginine or poly-L-histidine.

[0265] The composition for its use according to the invention may be in liquid form or in 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 lyophilization. When the composition is in liquid form and comprises micelles and is lyophilized in solid form, the micelles reform when the composition in solid form is again placed in an aqueous solution. The composition for its 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 of injectable solutions in humans or animals. It may be, for example, acids or bases to adjust the pH or NaCl to adjust the osmolarity.

[0266] The composition for its use according to the invention, in particular composition C1, is intended to be administered to humans or to animals and is therefore in a form suitable for such administration. When it is in liquid form, it is preferably suitable for administration by subcutaneous or intravenous route, in particular by intravenous infusion, and it 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 insulin pumps.

[0267] When it comes in solid form, it is preferentially suitable for administration:

[0268] - by transcutaneous route and it is preferably formulated and packaged in the form of a patch, or

[0269] - nasally in powder form, or

[0270] - sublingually in powder or tablet form, or

[0271] - by transmucosal route and it is preferably formulated and packaged in the form of a mucoadhesive tablet.

[0272] Manufacturing process

[0273] The useful composition according to the invention can be manufactured by any suitable process.

[0274] If the molecules that make up the composition are used as they are in a solvent, they can be mixed all together in said solvent.

[0275] If the molecules in the composition are conjugated to polymers and optionally in micelles for others, the manufacturing process may include the following steps:

[0276] - a. create an amphiphilic premix: mix in an aqueous solution:

[0277] - one or more Butyrate-Poly-L-Lysine conjugates

[0278] - one or more Lactate-Poly-L-Lysine conjugates

[0279] - one or more Propionate-Poly-L-Lysine conjugates - b. optionally, adding to the amphiphilic premix at least farnesylcysteine ​​and cholesterol, and stirring so as to form micelles formed by one or more of the conjugates of the amphiphilic premix encapsulating the farnesylcysteine ​​and cholesterol.

[0280] Preferably, the manufacturing method may comprise the following steps:

[0281] - a. create an amphiphilic premix: mix in an aqueous solution:

[0282] - one or more Butyrate-Poly-L-Lysine conjugates

[0283] - one or more Lactate-Poly-L-Lysine conjugates

[0284] - one or more Propionate-Poly-L-Lysine conjugates, and at least one or more conjugates chosen from:

[0285] - one or more Oleyl-Poly-L-Lysine conjugates

[0286] - one or more Palmitic-Poly-L-Lysine conjugates

[0287] - one or more Lauryl-Poly-L-Lysine conjugates

[0288] - one or more Azéayl-Poly-L-Lysine conjugates

[0289] - one or more Palmitoleyl-Poly-L-Lysine conjugates

[0290] - one or more Thioctyl-Poly-L-Lysine conjugates

[0291] - one or more Myristyl-Poly-L-Lysine conjugates

[0292] - one or more Orotyl-Poly-L-Lysine conjugates

[0293] - one or more Acetate-Poly-L-Lysine conjugates

[0294] - one or more linoleyl-Poly-L-Lysine conjugates, and

[0295] - b. adding to the amphiphilic premix at least farnesylcysteine ​​and cholesterol, and stirring so as to form micelles formed by one or more of the conjugates of the amphiphilic premix encapsulating the farnesylcysteine ​​and cholesterol.

[0296] A variation of the invention is to take advantage of the amphiphilic nature of most of the individual components to create an amphiphilic premix that allows for the controlled solubilization of the 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 the necessary time for their dissolution under stirring.

[0297] Poly-L-lysine may be replaced by another poly-lysine or by polyethylene glycol, poly-L-ornithine, poly-L-arginine or poly-L-histidine. Preferably, stirring is carried out for at least 60 minutes, even more preferably between 5 and 20 minutes, and preferably at a stirring speed of less than or equal to 900 revolutions per minute, in particular at a stirring speed of between 50 and 800 revolutions per minute.

[0298] According to a preferred embodiment, the manufacturing process according to the invention also comprises a step c. of separation of 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 physical separation process (filtration, ultrafiltration) is preferentially carried out to ensure the isolation of the soluble fraction, containing both the amphiphilic premix and the solubilized molecules.

[0299] 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.

[0300] 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 the farnesylcysteine ​​and cholesterol.

[0301] The 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 depending on 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.

[0302] Examples of the realization of amide, urea and carbamate bonds are described for example in:

[0303] - 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-ll (liaisons amide),

[0304] - 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 (liaisons amide),

[0305] - 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 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(l-Lysine Citramide Imide) Conjugates and Structure-dependence of the Drug Release, Journal of Drug Targeting, 12:5, 297-307, (liaisons carbamate),

[0306] -Elmore, W.M. (2013). Nanoparticles Stabilized with MPEG-Polylysine Carbamate: Synthesis and Characterization, (carbamate bonds),

[0307] - 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).

[0308] Examples:

[0309] Example of a composition suitable for use according to the invention:

[0310] Example 1

[0311] An example of a composition suitable according to the invention is presented below.

[0312] [Table 1]

[0313] 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 to 20 minutes depending on the solubility of the conjugate.

[0314] Example 2

[0315] An example of a composition suitable according to the invention is presented below.

[0316] [Table 2]

[0317] 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 to 20 minutes depending on the solubility of the conjugate.

[0318] Example 3 An example of a composition suitable according to the invention is presented below.

[0319] [Table 3]

[0320] 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 to 20 minutes depending on the solubility of the conjugate.

[0321] Example 4

[0322] An example of a composition suitable for testing on mice is presented below.

[0323] [Table 4]

[0324] 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 to 20 minutes depending on the solubility of the conjugate.

[0325] Example 5

[0326] An example of a composition suitable according to the invention is presented below.

[0327] [Table 5] 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 to 20 minutes depending on the solubility of the conjugate.

[0328] Example 6: Example of composition Cl An example of composition Cl useful according to the invention is presented below.

[0329] The following amounts 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. [Table 6]

[0330] 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 to 20 minutes depending on the solubility of the conjugate.

[0331] 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.

[0332] [Table 7] In a sterile 500 ml flask equipped with a magnetic stir bar, 23.8 ml of each previously prepared conjugate solution was added under magnetic stirring (700 rpm). The corresponding amounts of salts (previously lyophilized) in 300 ml of PBS were added to the conjugate solution under magnetic stirring (700 rpm). Cholesterol and farnesylcysteine ​​were added and the mixture was left stirring for 25 hours (700 rpm).

[0333] 30 ml of Cl 10 x formulation was withdrawn from a laminar flow hood and added to a new sterile 500 ml bottle.

[0334] 270 ml of sterile PBS was added to obtain the Cl lx formulation. The formulation was stirred for 30 minutes.

[0335] Example 7: Example of Cl composition

[0336] An example of a Cl composition suitable for testing on mice is presented below.

[0337] The following amounts 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.

[0338] [Table 8] 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 to 20 minutes depending on the solubility of the conjugate.

[0339] 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.

[0340] [Table 9]

[0341] In a sterile 500 ml flask equipped with a magnetic stir bar, 23.8 ml of each previously prepared conjugate solution was added under magnetic stirring (700 rpm). The corresponding amounts of salts (previously lyophilized) at 300 of PBS were added to the conjugate solution under magnetic stirring (700 rpm).

[0342] Cholesterol and farnesylcysteine ​​were added and the mixture was allowed to stir for 25 hours (700 rpm).

[0343] 30 ml of Cl 10 x formulation was withdrawn from a laminar flow hood and added to a new sterile 500 ml bottle.

[0344] 270 ml of sterile PBS was added to obtain the Cl lx formulation. The formulation was stirred for 30 minutes.

[0345] The vials were prepared according to the following plan using a sterile, calibrated 5 ml pipette:

[0346] Cl IX = 83 vials, 2 ml each.

[0347] Cl 10X = 83 vials of 2 ml each.

[0348] The 166 vials were placed in a steel freeze-drying tank and a one-day freeze-drying cycle was started.

[0349] Example 8: Molecular Analysis of the Fecal Microbiota of Mice (Wild WT, SOD1, SOD1 + composition according to the invention).

[0350] 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. In this test, 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.

[0351] [Table 10]

[0352] Feces from five male and female mice from each group (WT, SOD1, SOD1 + low dose Cl and SOD1 + high dose Cl), collected from the cages, were analyzed at TO (=T6), T3 weeks (T9) and 10 weeks (T16) for a total of 55 samples.

[0353] 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 migration of the samples on agarose gel. The DNA concentration was then determined using Nanodrop technology.

[0354] The obtained gDNA samples were then sequenced. The approach used is high-throughput sequencing of the gene coding for 16S ribosomal RNA (V3-V4 region) using Illumina technology (read length: 150 bases, read depth: 1 million reads).

[0355] The taxonomic affiliation 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.

[0356] 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 the sequences, whether for estimating the abundance of bacterial phyla / families or for performing statistical analyses (composition, diversity analyses, differential abundance analyses).

[0357] The comparison of bacterial diversity and richness between microbiota from treated and untreated mice was carried out by calculating the Shanon 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 packages.

[0358] Differential OTU abundance analyses were performed using the normalization and estimation tools of the DESeq2 package [Love, MI, Huber, W., Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 Genome Biology 15(12):550; 2014],

[0359] Correlation Microbiota - Clinical Parameters

[0360] 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).

[0361] Several analysis methods were tested, based on rank correlation (spearman).

[0362] The method used is the canonical correlation analysis (rcca) 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(ll):el005752). This method allows cross-validation of the results of analyses of correlations between the abundance of OTUs and the different clinical parameters measured. The results obtained are presented in the form of a heat map showing the significant correlations, positive or negative, between the abundance of OTUs and the clinical parameters.

[0363] Results :

[0364] Comparison of the composition of the fecal microbiota of KO “S0D1” mice with that of “

[0365] Wild Type » (WT)

[0366] Analysis of the relative abundance of bacterial phyla, orders, families and even genera present in SD01 mice compared to WT mice shows that the composition of the S0D1 microbiota differs from that of WT:

[0367] - S0D1 have fewer Firmicutes and more Bacteroidetes than WT - Among Bacteroidetes, SOD1 mice harbor more Bacteroidales, especially Muribaculaceae, and more Porphyromonaceae, including Muribaculum, than WT

[0368] - Among Firmicutes, the relative abundance of Lachnospiraceae (Shaedle re Ha 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.

[0369] 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 made it possible to determine that there is:

[0370] - No significant difference in bacterial diversity (indices: Chaol, Observed, Shannon and Inv Simpson) between SOD1 and WT (alpha diversity).

[0371] - A significant difference (Jaccard: p= 0.01 and Bray Curtis: p=0.01) in bacterial species richness (beta diversity). No effect of time (TO, T6 and T16) was found.

[0372] The Jaccard and Bray Curtis index or distance are two well-known metrics for measuring similarity, dissimilarity, and diversity between multiple samples.

[0373] Finally, PLS-DA analysis shows that the fecal microbiotas of WT and SOD1 mice differentiate (Figure 1).

[0374] In conclusion, SOD1 mice have a disturbed intestinal microbiota compared to WT wild-type mice, characterized by lower bacterial richness, a higher proportion of Gram- bacteria (Muribaculaceae and Muribaculum) as well as Gram- bacteria Lactobacillales and Eggerthelaceae but a lower proportion of Lachnospiraceae and Erysipelotrichales.

[0375] Result: Effect of administration of composition Cl (associated with C2) at high or low dose in SOD1 mice

[0376] A - Administration of a high dose 10X of Cl (associated with C2)

[0377] The compositions Cl and C2 are described in Tables 7 to 10.

[0378] Administration of composition Cl (associated with C2) at a high dose for 3 weeks results in:

[0379] - An increase in the relative abundance of:

[0380] *Bacteroidales especially Muribaculaceae

[0381] * Oscillospiraceae

[0382] *COE1 - A decrease in the abundance of:

[0383] * Lactobacillaceae (Lactobacillus genus)

[0384] * Lachnospiraceae

[0385] Administration of a high dose of the Cl composition for 3 weeks appears to accentuate the disturbances observed in the SOD1 mouse.

[0386] B - Administration of a low dose of Cl IX (associated with C2)

[0387] 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:

[0388] - An increase in the relative abundance of:

[0389] * Lachnospiraceae

[0390] * Oscillospirals

[0391] * GAC-485

[0392] - A decrease in the relative abundance of:

[0393] *Lactobacillale

[0394] The low dose appears to partially rebalance the intestinal microbiota of SOD1 mice to bring it closer to 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 marked after 10 weeks of treatment than after 3 weeks.

[0395] Result: Comparison of bacterial diversity and richness in treated and untreated SOD1 mice.

[0396] Analysis of the alpha diversity of fecal microbiota did not reveal any significant effect of Cl administration (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) (Figure 2 and 3).

[0397] Beta diversity analysis showed significant differences between untreated SOD1 mice and treated SOD1 mice.

[0398] - Controls versus Cl (associated with C2) high dose (p=0.02 Jaccard and p= 0.01 Bray curtis)

[0399] - Cl (associated with C2) low dose versus Cl (associated with C2) high dose (p = 0.02 Jaccard and p = 0.01 Bray Curtis)

[0400] - No significant difference between untreated control and Cl (associated with C2) low dose. Results: reason between sa mice and SOD1 mice treated or not with different doses of Cl (associated with C2)

[0401] PLS-DA analysis of the composition of all the microbiotas studied showed that these microbiotas could be separated into 4 distinct groups, the microbiota of mice treated with the low dose of Cl (associated with C2) approaching, in the same cluster, the microbiota of wild mice (Figure 4).

[0402] Conclusion

[0403] The study showed that the microbiota composition 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 since the relative abundance of certain major bacterial groups is altered. It is characterized, in particular, by a higher abundance of Muribaculaceae and lactobaci I la les (Oscillospiraceae) to the detriment of Lachnospiraceae and Erysipelotrichales. PLS-DA analysis further demonstrated that the microbiotas of wild-type and SOD1 mice were separate.

[0404] Administration of a high dose of Cl (associated with C2) to SOD1 mice for 3 weeks appears to amplify the differences observed between wild-type mice and SOD1 mice (increase in Muribaculaceae and Oscillospiraceae and decrease in Lachnospiraceae). The composition of the microbiota of SOD1 mice treated with the high dose of Cl (associated with C2) is very clearly different from that of SOD1 controls but also of wild-type mice, this group of mice treated with the high dose being largely different from the others (most disturbed microbiota).

[0405] Administration of a low dose of Cl (associated with C2) to SOD1 mice for 3 to 10 weeks appears to partially rebalance the microbiota disturbances observed in control SOD1 mice. Indeed, an increase in Lachnopiraceae and a decrease in Lactobacillales, microbial groups affected in SOD1 mice compared to wild-type mice, is observed. The microbiota composition of SOD1 mice treated with a low dose of Cl (associated with C2) thus approaches that of wild-type mice, with the microbiotas of these two groups being grouped in the same cluster.

[0406] 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) indeed appears to have a beneficial effect on the disturbed microbiota of SOD1 mice.

[0407] 9: Correlation study between and molecular composition of the microbiota in SOD1 mice after administration of a low dose of Cl (associated with

[0408] The objective of the study was to analyze potential correlations between the molecular composition of the intestinal 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.

[0409] This study was carried out in female mice whose molecular analysis of the fecal microbiota had been previously performed (five females from each group [WT, SOD1, SOD1- low dose), collected in the cages, at TO (=T6), T3 weeks (T9) and 10 weeks (T16)].

[0410] Tests were performed at least 1 h after the first daily dose of test compound or vehicle.

[0411] A one-day session included a 5-min training trial at 4 RPM on the rotating apparatus (AccuScan Instruments, Columbus, USA). One hour later, animals were tested for 3 consecutive 6-min acceleration trials with the speed increasing from 0 to 40 RPM over 360 s and an inter-trial interval of at least 30 min. The latency to fall off the rod was recorded

[0412] Correlation study in SOD1 and Wild Type mice.

[0413] RCCA (Regularized Canonical Correlation Analysis) correlation analysis mainly showed in WT and SOD1 mice the existence of a significant negative correlation between the clinical score, the distance traveled and the 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 the recovery time (Figure 5).

[0414] Although the relative abundance of the Desulfovibrionaceae family (like that of Prevotellaceae) is low compared to other bacterial families, we were nevertheless able to find the presence of OTUs of this family in SOD1 mice only (no detection in Wild Type mice). This could suggest a potential role of Desulfovibrionaceae in the pathophysiology in SOD1 mice.

[0415] Correlation study in SOD1 mice treated or not treated with the low dose of the Cl composition (associated with C2) for 10 weeks

[0416] The RCCA correlation analysis carried out in control SOD1 mice and those 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 is, the greater the distance traveled by the mice. Knowing that one of the main effects of a composition according to the invention on the intestinal 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 (Figure 6).The evidence of a negative correlation between Desulfovibrionaceae and the severity of symptoms in SOD1 mice suggests that species of this family, which produce sulfides, 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, ulcerative colitis, etc.). At the same time, the lower abundance of species of the Lachnospiraceae family in SOD1 mice could result in a decrease in intracolonic butyrate concentration, as a large proportion of Lachnospiraceae species produce this metabolite with proven health effects.

[0417] The administration of a composition according to the invention at a low dose for 10 weeks is accompanied by changes in the composition of the microbiota of the SOD1 mouse, resulting, in particular, in an increase in the abundance of Lachnospiraceae and the disappearance of Desulfovibrionaceae. This correlates with the significant improvement in the ability of the mice 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, causing metabolic modifications 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 an anhydride of lactic acid, -butyric acid, and / or a salt and / or an ester and / or an 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 the disease of Charcot, 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 any one of the preceding claims, as a medicinal product or food supplement in 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 Desulfovibrionaceae 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, polyornithine, polyarginine and polyhistidine.

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, poly-ornithine, poly-arginine and poly-histidine.

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 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.

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 an 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, polyornithine, 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 its 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 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 farnesylcysteine ​​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 farnesylcysteine ​​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 any one of claims 20 or 21, the composition being characterized in that Poly-L-Lysine is replaced by another poly-lysine or by polyethylene glycol, poly-L-ornithine, 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.

Images