Carbon monoxide releasing molecules (CORM) or compositions thereof for treating or preventing intestinal flora disorders in subject

Abstract

The present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in the treatment or prevention of intestinal flora disorders in a subject. More specifically, the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in the treatment or prevention of intestinal dysbiosis, preferably cecum and / or colon dysbiosis, in a subject wherein the carbon monoxide releasing molecule or the composition thereof is administered orally.

Description

Technical Field

[0001] The present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for treating or preventing intestinal dysbiosis in a subject. Background Art

[0002] The mammalian gastrointestinal microbiota consists of 10 12 Up to 10 14 Microorganisms, such as bacteria, viruses or even eukaryotic organisms (such as fungi or yeast), that colonize and survive in the digestive tract of mammals shortly after birth [1]. The gastrointestinal microbiota is mainly located in the small intestine and colon, distributed between the lumen of the digestive tract and the protective biofilm formed by the intestinal mucus that covers its inner wall [2]. Due to the acidity of the stomach, the stomach has 100 million times fewer commensal bacteria than the colon. For this reason, most of the time it is called the gut microbiota or intestinal microbiota.

[0003] The human intestinal microbiota is gradually constructed from birth, coming into contact with maternal and environmental bacteria at birth, and continues to multiply through feeding and other contacts, followed by gradual bacterial colonization in a specific order. It is certain that the bacterial components of the human intestinal microbiota are mainly composed of Firmicutes, Bacteriodetes, Actinobacteria, Proteobacteria, Fusobacteria, and Archaea, among which Firmicutes and Bacteroidetes account for the majority [3].

[0004] The use of high-throughput sequencing methods has made it possible to characterize all the microbial genomes found in the intestine and identify a thousand different species, of which bacteria make up the majority.

[0005] The gut microbiota is unique in quality and quantity to each person with some common features. Of the 160 bacterial species found on average in the microbiota of healthy individuals, only half are found in common between individuals. However, there are 15 to 20 common bacterial species present in all people that are responsible for the basic functions of the microbiota [2].

[0006] The first intestinal bacteria to colonize the intestine are aerobic bacteria such as Enterococci, Staphylococci, etc., which require oxygen to proliferate. By consuming the oxygen present in the intestine, aerobic bacteria then promote the implantation of anaerobic bacteria such as Bacteroides, Clostridium, Bifidobacterium, Lactobacillus, etc., which do not require oxygen to proliferate, or in some cases only proliferate in the absence of oxygen.

[0007] During the first few years of life, the composition of the gut microbiota evolves in quality and quantity, depending on factors such as dietary diversity, genetics, hygiene levels, medical care received and environment. This composition remains remarkably stable, although this stability appears to vary from person to person.

[0008] There are more than 10 14 Microorganisms cover the entire surface of the digestive tract, primarily in the intestines, where the intestinal microbiota acts as a barrier against invading agents by competing with pathogens for nutrients and binding sites, producing inhibitory substances, or preventing pathogens from entering the intestinal mucosa. In addition, the microbial genome encodes 3 to 4 million genes (approximately 150 times the human genome), enabling microorganisms of the microbiota to perform several metabolic activities that are not encoded by the human genome [1].

[0009] There is a mutually beneficial dynamic between the microorganisms in the gut microbiota and the host organism, which allows the maintenance of normal immune, metabolic and motility functions and even proper nutrient digestion and absorption [4].

[0010] Considering the importance of a healthy gut microbiota in maintaining various biological functions, gut dysbiosis (meaning alterations in the quantity and / or quality and / or function of the gut microbiota) may cause or contribute to various physiological problems or diseases.

[0011] During life, several factors may influence the composition of the intestinal microbiota to a greater or lesser extent and cause or at least favor intestinal dysbiosis. Among these factors, one can list aging, various diseases, medical treatments, lifestyle or dietary changes, or even the presence of pesticides or additives in food.

[0012] Gut dysbiosis refers to an imbalance in the gut microbial community that results in alterations in the quantity and / or quality and / or function of the gut microbiota. Considering the importance of a healthy gut microbiota for maintaining various physiological functions, it is crucial to maintain a healthy gut microbiota throughout life or to be able to restore a healthy gut microbiota after temporary dysbiosis.

[0013] Prebiotic and probiotic compositions are known and are often proposed for maintaining or restoring the intestinal microbiota. Probiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host, whereas prebiotics are generally non-digestible, fermentable ingredients that induce specific changes in the composition and / or activity of the resident microflora by selectively increasing the proliferation of bacteria that are responsible for the beneficial effects on host well-being and health [5][6].

[0014] Interestingly, the inventors have proposed a novel method for restoring or maintaining a subject's intestinal microbiota to a healthy state. Thus, the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for treating or preventing intestinal flora imbalance in a subject (preferably a human).

[0015] In fact, the inventors have shown that oral administration of carbon monoxide-releasing molecules (CORMs) or compositions thereof to a subject can increase and / or decrease the abundance of one or more bacterial species, particularly anaerobic bacterial species, present in the subject's intestinal microbiota relative to the abundance of bacterial species present in the subject's intestinal microbiota before administration of the CORMs or compositions thereof.

[0016] These results are surprising as they demonstrate the ability of carbon monoxide releasing molecules (CORMs) or compositions thereof to pass through the gastrointestinal tract and specifically reach and accumulate in the intestine where they produce beneficial effects on the intestinal microbiota.

[0017] Various studies have been conducted to understand the role of carbon monoxide in many physiological and pathological processes and to determine the potential use of carbon monoxide-releasing molecules in the treatment or prevention of some physiological dysfunctions or even diseases. However, this is the first time that carbon monoxide-releasing molecules (CORMs) or compositions thereof are proposed for the treatment or prevention of intestinal flora imbalance in a subject (preferably a human). Summary of the invention

[0018] As mentioned above, interestingly, the inventors have proposed a new method for restoring or maintaining the composition of the intestinal microbiota of a subject to a healthy state. Interestingly, the inventors have shown that oral administration of carbon monoxide releasing molecules (CORMs) makes it possible to reconstruct the intestinal microbiota to a healthy phenotype, in particular by increasing and / or decreasing the abundance of some specific bacterial species present in the intestinal microbiota.

[0019] Therefore, the first object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for treating or preventing intestinal flora imbalance in a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally or rectally.

[0020] A second object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in a method for treating or preventing intestinal dysbiosis in a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally.

[0021] The third object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in a method for restoring or maintaining a healthy state of the intestinal microbiota of a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Shown are the levels of blood carboxyhemoglobin (COHb) over 48 hours after oral administration of CORM-401 to mice fed a high-fat diet. Statistical analysis was performed using one-way ANOVA followed by Fisher's multiple comparison test. Values ​​are expressed as mean ± SEM. *p < 0.01 vs. time 0.

[0023] Figure 2 The time-dependent CO accumulation in gastrointestinal tissues such as cecum (A) and colon (C) and cecal contents (B) and feces (D) of HFD mice orally administered CORM-401 or PBS (as control) is shown. Statistical analysis was performed by Student's t-test. Values ​​are expressed as mean ± SEM. *p < 0.01 vs. control. DETAILED DESCRIPTION

[0024] definition

[0025] Carbon monoxide (CO) is known to be an air pollutant, but is also an endogenous signaling molecule that can play a key role in many physiological and pathological processes. CO is produced endogenously in many organisms due to the breakdown of heme by heme oxygenases, which exist in constitutive (HO-2 and HO-3) and inducible (HO-1) isoforms. Most of the CO is bound to hemoglobin to form COHb, while the rest is distributed in tissues, exerting a variety of physiological functions, such as vasodilation, anti-inflammatory effects, and antiproliferative effects. Given the difficulty in storing CO and controlling its dosage for therapeutic use, various CO-releasing molecules have been developed [7].

[0026] The term "carbon monoxide releasing molecule (CORM)" refers to a compound that is intended to release controlled amounts of carbon monoxide (CO). CORMs are generally divided into metal CORMs (also called "metal carbonyl CORMs") and non-metal CORMs. CORMs can release carbon monoxide spontaneously, or by contact with, for example, a suitable solvent or medium, such as an aqueous physiological fluid (e.g., blood or lymph) or an aqueous physiological cellular material (e.g., tissue, organ, or cell). CORMs can also release carbon monoxide by irradiation. For example, the CORM can be irradiated before administration to produce a solution in which CO is dissolved, or irradiated in situ after administration.

[0027] Metal CORMs are compounds containing transition metals (e.g., nickel, cobalt, ruthenium, manganese, molybdenum, rhodium, boron, or iron) [8] surrounded by carbonyl (CO) groups (as coordinating ligands) and capable of releasing CO in biological systems. Metal CORMs may also contain other ligands that can modulate specific properties of the CORM, such as the rate of carbon monoxide release, solubility, hydrophobicity, stability, or electrochemical potential. Other ligands may be, for example, halides, sulfoxides, natural and synthetic amino acids, aromatic compounds, carboxylates, ethers, alcohols, or nitriles. Metal CORMs include carbon monoxide myoglobin (CO-Mb), carbon monoxide hemoglobin (CO-Hb), and polyethylene glycol-coupled hemoglobin (CO-MP4).

[0028] Non-metallic CORMs are metal-free carbon monoxide-releasing molecules with different chemical structures, among which borane carbonates (e.g., CORM-A1), CO donors based on the Diels-Alder reaction, 3-hydroxyflavone (3-HF), quinolones, cyclodiketones, xanthene-9-carboxylic acids, meso-carboxy BODIPY derivatives, etc. [7] Non-metallic CORMs include organic CO-releasing molecules such as oxalic acid (CAS No.: 144-62-7), cyclopentenone derivatives, cyclopentadienone-alkyne pairs, and S-arylthioformates.

[0029] In the context of the present invention, the CORM may be a salt, such as a pharmaceutically acceptable salt.

[0030] The term "pharmaceutically acceptable salt" refers to salts of acids or bases known to be used in the preparation of active ingredients for treatment. Examples of pharmaceutically acceptable acids suitable as anion sources are those disclosed in Handbook of Pharmaceutical Salts: Properties, Selection and Use (PH Stahl and CG Wermuth, Weinheim / Zürich: Wiley-VCH / VHCA, 200). Salts are salts for animals and humans that are approved by federal regulatory agencies or state governments or listed in the U.S. or European Pharmacopoeia or other generally recognized pharmacopoeias. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, 6-hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, sucrose, stearate, succinate, tannate, tartrate, toluenesulfonate, trifluoroacetate, and xinofoate. Suitable basic salts are formed from bases which form non-toxic salts. Examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts. Hemisalts of acids and bases can also be formed, for example hemisulphate and hemicalcium salts.

[0031] As used herein, the term "microbiota" (or also referred to as "microflora") refers to a community of microorganisms such as bacteria, archaea, protists, fungi and viruses that are typically present in an organ or part of the body.

[0032] The intestinal microbiota corresponds to the community of living microorganisms that inhabit the intestinal tract (mainly the small intestine and colon, including the cecum connecting the colon and small intestine).

[0033] The "cecum" corresponds to the first part of the colon. It consists of a pouch into which the opening of the ileo-cecal valve leads. It is at this valve that the small intestine (especially the ileum) empties into the colon. The term "colon" refers to the longest part of the large intestine, which is located between the cecum and the rectum. In the human cecum and colon, 10 12 CFU / mL or more of bacteria, mainly anaerobic bacterial species.

[0034] The term "anaerobic bacterial species" refers to a bacterial species that is sensitive to oxygen. These terms include: (i) strict anaerobic bacterial species, which are extremely sensitive to oxygen and cannot come into contact with oxygen in order to grow; (ii) aerotolerant anaerobic bacterial species, which can be exposed to low concentrations of oxygen without stopping their growth; and (ii) facultative anaerobic bacterial species, which are able to grow in the presence or absence of oxygen.

[0035] The term "aerobic bacterial species" refers to bacterial species that survive and grow only in the presence of oxygen in the environment.

[0036] The term "dysbiosis" corresponds to a physiological state in which a subject exhibits a microbial profile that is different from or significantly deviates from the corresponding microbial profile typically possessed by a normal (healthy) subject, such as a change in the diversity and / or number of commensal species of the microbial population compared to a normal (healthy) subject. Dysbiosis can be diagnosed and the extent of dysbiosis measured by comparing how the microbial profile differs from that of a normal (healthy) subject. A typical microbial profile of a normal (healthy) subject can be obtained from a single subject or even a single sample of a single subject, but preferably from multiple subjects, and the techniques used allow for intra-individual variation. The comparison between the profile of a test sample and the profile of a normal (healthy) reference for assessing whether a test sample is dysbiotic and optionally the extent of any dysbiosis can be achieved by any convenient means, and the choice of the method used can be determined by the form of the data constituting the profile to be compared. Depending on the nature of the data, the comparison can be a qualitative, semi-quantitative or quantitative process. There are various methods to determine the presence of dysbiosis, especially by different indicators based on comparison with a group of individuals or samples used as a reference.

[0037] The term "intestinal dysbacteriosis" refers to an imbalance in the intestinal microbial community that causes changes in the quantity and / or quality and / or function of the intestinal microbial community compared to the corresponding microbial community of a normal (healthy) subject. Conveniently, the dysbacteriosis index test described in WO2016 / 156251 can be used to assess the intestinal dysbacteriosis state of a sample from a test subject. The test can be compared with the intestinal microbial community spectrum of a healthy subject to analyze the intestinal microbial community spectrum of the test sample, and a relative score proportional to the degree of dysbacteriosis in the sample is applied. Therefore, the sample can be classified as dysbacteriosis or not, and the degree of any dysbacteriosis can also be determined. Therefore, before, during and / or after treatment, changes in the dysbacteriosis / normal state of the subject can be easily monitored.

[0038] The term "cecal and / or colonic dysbacteriosis" refers to an imbalance in the cecal and / or colonic microbial community resulting in an altered quantity and / or quality and / or function of the cecal and / or colonic microbiota compared to the corresponding microbiota of a control subject. The control subject may correspond to a normal healthy subject, or to a subject without the same disease as the subject suffering from cecal and / or colonic dysbacteriosis.

[0039] The terms "treating" or "treatment" mean to reverse, alleviate, inhibit or slow the progression of the disease or condition to which such term applies or one or more symptoms of such disease or condition in a subject.

[0040]

[00136] The terms "preventing" or "prevention" mean reducing the risk of a subject developing the disease or condition to which such term applies, or one or more symptoms of such disease or condition.

[0041] The terms "microbiota health state", "microbiota normal state" or "healthy microbiota state" correspond to the typical profile of the microbiota of a normal healthy subject, which may also be referred to as "normobiosis" or "normobiotic state". A "microbiota health state" corresponds to a microbiota in which the microbiota number and / or diversity corresponds to the microbiota number and / or diversity of a healthy subject (meaning that it does not suffer from dysbiosis).

[0042] The term "healthy subject" or "normal subject" refers to a subject that does not suffer from intestinal dysbiosis.

[0043] As used herein, the terms "subject", "patient" or "individual" are used interchangeably and refer to humans, non-human mammals (e.g., rodents (mouse, rat), felines, canines or primates), birds and fish that have or may have intestinal dysbiosis. Preferably, the subject is a human, male or female.

[0044] The term "high fat diet" refers to a diet consisting of at least 35% of the total calories consumed from unsaturated and saturated fats.

[0045] The term “abundance” refers to the amount of a microorganism to which the term applies. The abundance of microorganisms in the gut microbiota can be measured by extracting DNA and RNA from fecal samples and then sequencing them using metagenomic / metatranscriptomic techniques, as previously described in the literature [9].

[0046] The term "pharmaceutically acceptable" means approved by a federal regulatory agency or a state government or listed in the U.S. or European Pharmacopeia or other generally recognized pharmacopeia for use in animals and humans.

[0047] The term "pharmaceutical composition" refers to a composition comprising a pharmaceutically acceptable carrier, and a "nutraceutical composition" refers to a composition comprising a physiologically acceptable carrier. For example, a carrier can be a diluent, adjuvant, excipient or vehicle for administering a therapeutic agent. Such a carrier can be a sterile liquid, such as water and oil, including oils from petroleum, animal, plant or synthetic sources, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Suitable pharmaceutical / physiological excipients include starch, glucose, lactose, sucrose, sodium stearate, glyceryl monostearate, talc, sodium chloride, dry skim milk, glycerol, propylene glycol, water, ethanol, etc. In the context of the present invention, a pharmaceutical composition or a nutraceutical composition is suitable for oral administration. For example, tablets or capsules can be prepared by conventional methods with pharmaceutically acceptable excipients, such as binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silicon dioxide); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). Tablets can be coated by methods well known in the art. For example, liquid preparations for oral administration can take the form of, for example, solutions, syrups, or suspensions, or can be presented as a dry product for constitution with water or other suitable carriers prior to use. Such liquid preparations can be prepared by conventional methods with pharmaceutically acceptable additives such as suspending agents (such as sorbitol syrup, cellulose derivatives or hydrogenated edible fats), emulsifiers (such as lecithin or gum arabic), non-aqueous carriers (such as almond oil, oily esters, ethanol or fractionated vegetable oils) and preservatives (such as methyl paraben or propyl paraben or sorbic acid). The preparation can also contain buffer salts, flavoring agents, coloring agents and sweeteners as required. According to the present invention, pharmaceutical compositions or nutritional compositions can be capsules, tablets, powders, pills, dragees, granules, sachets, gels, pastes, syrups, emulsions, suspensions, suppositories, solutions, etc.

[0048] The term "nutraceutical" refers to a common food to which components or ingredients are added that may make it have a specific medical or physiological benefit different from a pure nutritional effect. It includes dietary supplements and functional foods. It can be in liquid form, solid form, powder form, such as pills, tablets, capsules, granules, gels, pastes, syrups, emulsions, suspensions, solutions, beverages, decoctions, nutrition bars, confectionery, dairy products or fermented dairy products, yogurt, dairy powders, enteral nutrition products, infant and / or infant compositions, fermented or non-fermented cereal-based products, ice cream, chocolate, coffee, "culinary" products such as mayonnaise, ketchup or salad dressing.

[0049] The present invention

[0050] The first object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for treating or preventing intestinal flora imbalance in a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally or rectally.

[0051] The second object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in a method for treating or preventing intestinal flora imbalance in a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally.

[0052] The third object of the present invention relates to a carbon monoxide releasing molecule (CORM) or a composition thereof for use in a method for restoring or maintaining a healthy state of the intestinal microbiota of a subject, wherein the carbon monoxide releasing molecule or the composition thereof is administered orally.

[0053] A fourth object of the present invention relates to the use of CORM or a composition thereof for increasing the abundance of potentially beneficial bacterial species present in the intestinal microbiota of a subject, such as one or more bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Alistipes shahii, Olsenella umbonata and Olsenella uli.

[0054] The term "restoration" or "reshaping" includes any positive effect on the intestinal dysbiosis of a subject. Thus, not only is complete normalization of the intestinal microbiota profile (i.e. restoration of normobiosis or "normalized" or normal intestinal microbiota profile) covered, but also partial improvement of the intestinal microbiota profile of a subject or even improvement of the intestinal microbiota profile is covered. The improvement may be partial in the sense that the perturbation of the abundance or metabolic phenotype of a subpopulation or microbiome of the profile is improved (preferably normalized) and / or the degree of perturbation of the abundance or metabolic phenotype of a specific microorganism or microbiome is partially reduced. Preferably, the intestinal microbiota of a subject suffering from intestinal dysbiosis is (completely) normalized.

[0055] The term "maintain" includes that a normal intestinal microbiota profile generally continues to exist in a subject or part thereof during administration of an effective amount of a CORM or composition thereof. Thus, there is no significant disturbance in the abundance or metabolic phenotype of one or more microorganisms or microbial groups.

[0056] In some embodiments, the CORM is selected from:

[0057] - Metal carbonyls, such as CORM-401 (CAS No. 1001015-18-4), CORM-2 (CAS No. 22594-69-0), CORM-3 (CAS No. 475473-26-8), CORM-371, EBOR-CORM-1, ALF-186, ALF-850 and ALF-021, carbon monoxide myoglobin (CO-Mb), carbon monoxide hemoglobin (CO-Hb) and polyethylene glycol-coupled hemoglobin (CO-MP4);

[0058] - metal-free CO-releasing compounds, such as CORM-A1 (CAS No. 17363-08-5); and / or

[0059] - Organic CO-releasing molecules, such as oxalic acid (CAS No. 144-62-7), cyclopentenone derivatives, cyclopentadienone-alkyne pairs and S-arylthioformates.

[0060] Transition metal carbonyls EBOR-CORM1, CORM-371, ALF-186, ALF-850 and ALF-021[7] have the following chemical formulas and / or structures:

[0061] Table 1

[0062]

[0063]

[0064] For example, the cyclopentadienone-alkyne pair can be selected from: BW-CO-104, BW-CO-105, BW-CO-106, BW-CO-108, BW-CO-109, BW-CO-110, BW-CO-111, BW-CO-112 and / or BW-CO-113, as disclosed in Zhixiang Pan et al., "Organic CO-prodrugs: Structure CO-release rate relationship Studies", Chem. Eur. J. 10.1002 / chem.201700936. The S-aryl thioformate can be selected from the list disclosed in DeSimone et al., “ThioCORMates: Tunable and Cost-Effective Carbon Monoxide-Releasing Molecules”, Chemistry, May 9, 2022. doi: 10.1002 / chem.202201326

[14] , more specifically selected from the S-thioformate compounds listed in Table 2 below.

[0065] These compounds have the following chemical formulas and / or structures:

[0066] Table 2

[0067]

[0068]

[0069] In some embodiments, the carbon monoxide releasing molecule is selected from CORM-401, CORM-A1, or a mixture thereof.

[0070] In some embodiments, the carbon monoxide releasing molecule is CORM-401 (CAS No: 1001015-18-4).

[0071] In some embodiments, the CORM refers to a chemically modified CORM, such as a CORM coupled to a copolymer, or a CORM modified to be sensitive to a particular enzyme, light radiation, or other stimulus.

[0072] In some embodiments, the carbon monoxide-releasing molecule is selected from carboxymyoglobin (CO-Mb), carboxyhemoglobin (CO-Hb), and polyethylene glycol-coupled hemoglobin (CO-MP4).

[0073] In some embodiments, the CORM is coupled to a copolymer such as polyethylene glycol. An example of a CORM coupled to polyethylene glycol is polyethylene glycol coupled hemoglobin, which is referred to as "CO-MP4".

[0074] In some embodiments, the subject is a human.

[0075] In some embodiments, the intestinal dysbiosis is cecal and / or colonic dysbiosis, preferably colonic dysbiosis.

[0076] Therefore, according to a specific embodiment of the present invention, CORM or a composition thereof is administered to a subject before, during and / or after a high-fat diet or a drug treatment that induces dysbiosis (e.g., antibiotic treatment). According to a preferred aspect, CORM or a composition thereof is administered to a subject at least during a high-fat diet or a drug treatment that induces dysbiosis (e.g., antibiotic treatment), preferably before, during and after a high-fat diet or a drug treatment that induces dysbiosis (e.g., antibiotic treatment).

[0077] For example, the antibiotic treatment can be selected from clindamycin, vancomycin, dalbavancin, oritavancin, telavancin, neomycin, sulfasalazine, ampicillin, phenoxymethylpenicillin, flucloxacillin, tobramycin, metronidazole, gentamicin, bacitracin, streptomycin, amoxicillin, cephalexin, lymecycline, tetracycline, deoxyoxytetracycline, ciprofloxacin, levofloxacin, norfloxacin, erythromycin, azithromycin, clarithromycin, sulfamethoxazole (cotrimoxazole) ole), furazolidone, trimethoprim, sulfamethoxazole, amikacin, erythromycin, fedamycin, gemifloxacin, moxifloxacin, delafloxacin, cefuroxime, ceftriaxone, ceftazidime, ceftaroline, cefotetan, cefotaxime, cefiderocol, cefepime, cefdinir, omadacycline, minocycline, eracycline, demeclocycline, sarecycline, imipenem, cilastatin, meropenem, etapenem. In some embodiments, the abundance of one or more bacterial species is increased and / or decreased relative to the abundance of the bacterial species before administration of the CORM or a composition thereof.

[0078] In some embodiments, the CORM or composition thereof used according to the present invention promotes or facilitates the restoration of a subject's intestinal microbiota to a normal healthy state. The CORM or composition thereof according to the present invention may reduce the abundance of potentially harmful bacterial species and / or increase the abundance of potentially beneficial bacterial species present in the intestinal microbiota compared to the abundance of these bacterial species before the CORM or composition thereof is administered to a subject suffering from intestinal dysbiosis.

[0079] In some embodiments, the CORM or a composition thereof reduces the risk of increased abundance of potentially harmful bacterial species and / or decreased abundance of potentially beneficial bacterial species present in the intestinal microbiota. In some embodiments, the CORM or a composition thereof used according to the present invention restores the intestinal microbiota of a subject to a healthy state.

[0080] In some embodiments, CORMs or compositions thereof used according to the present invention maintain the gut microbiota of a subject in a healthy state.

[0081] In more particular embodiments, the abundance of one or more bacterial species is normalized, or maintained to a normal abundance, relative to the abundance of the one or more bacterial species prior to administration of the CORM or composition thereof.

[0082] In some embodiments, the abundance of one or more bacterial species is increased or decreased by 2-fold, 3-fold, or 4-fold relative to the abundance of the bacterial species prior to administration of the CORM or composition thereof.

[0083] Methods for measuring the relative abundance of intestinal microbiota in a sample include, but are not limited to, nucleic acid analysis (e.g., nucleic acid sequencing methods, methods based on oligonucleotide hybridization probes, methods based on primers for nucleic acid amplification), methods based on antibodies or other specific affinity ligands, proteomics and metabolomics methods. Analysis of ribosomal RNA genes and their transcripts (e.g., 5S, 16S, and 23S genes (preferably 16S genes) in prokaryotic microorganisms or 18S genes in eukaryotic microorganisms) may be convenient.

[0084] In some embodiments, the bacterial species is one or more bacterial species present in the gut microbiota. For example, the bacterial species is selected from one or more of the following bacterial species: Akkermansia muciniphila, Alistipes fingoldii, Alistipes shahii, Olsenella umbonata, Olsenella uli, Faecalibaculum rodentium, Lactobacillus murinus, Lactobacillus fermentum, Libanicoccus massiliensis, Lactobacillus salivarius, Lactobacillus agilis, Lactobacillus ruminis, Lactobacillus salivary, Parabacteroides goldsteinii, Aerococcus viridans, Rombutsia ilealis, Lactococcus lactis, lactis, Enterococcus faecium, Streptococcus thermophiles, Enterococcus faecalis, Lactococcus piscium, Carnobacterium divergens, Listeria monocytogenes.

[0085] In some embodiments, the CORMs or compositions thereof used according to the present invention normalize or maintain the abundance of one or more, e.g., at least 2, 3, 4, 5 or all of these bacterial species, preferably in a human subject.

[0086] The bacterial species Akkermansia muciniphila, Alistipes fingoldii, Allelobacterium sabralskii, Olsenella umbonata, Erwinia gingivalis, Faecalibacterium rodentium, Lactobacillus murinus, Lactobacillus fermentum, Libanicoccus massiliensis, Lactobacillus salivarius, Lactobacillus agile, Lactobacillus rumenicus, Lactobacillus animalis, Parabacteroides archaeae, Aerococcus viride, and in particular the bacterial species Akkermansia muciniphila are among the most abundant bacterial species present in the gut microbiota of healthy normal subjects. The abundance of one or more of these bacterial species may be reduced in the gut microbiota of subjects with gut dysbiosis, especially in the setting of a high fat diet or drug treatment that induces dysbiosis.

[0087] In some embodiments, the abundance of one or more anaerobic bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Bacillus sabdariffa, Olsenella umbonata, Erwinia gingivalis, Faecalibacterium rodentium, Lactobacillus mouse, Lactobacillus fermentum, Libanicoccus massiliensis, Lactobacillus salivarius, Lactobacillus agile, Lactobacillus rumenica, Lactobacillus animalis, Parabacteroides archaeopteris, and Aerococcus viride is increased relative to the abundance of these anaerobic bacterial species described below before administration of the CORM or a composition thereof.

[0088] In specific embodiments, the abundance of one or more anaerobic bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Alternaria sabdariffa, Olsenella umbonata, and Erwinia gingivalis is increased relative to the abundance of these anaerobic bacterial species described below prior to administration of the CORM or a composition thereof.

[0089] In specific embodiments, the abundance of the anaerobic bacterial species Akkermansia muciniphila is increased relative to its abundance prior to administration of the CORM or composition thereof.

[0090] In the intestinal microbiota of subjects with intestinal dysbiosis, especially in the case of a high-fat diet or drug treatment inducing dysbiosis, the abundance of the bacterial species Rombutsia ilealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophiles, Enterococcus faecalis, Lactococcus piscium, Carnobacterium divergens, Listeria monocytogenes, especially the bacterial species Romboutsia ilealis is increased.

[0091] In some embodiments, the abundance of one or more anaerobic bacterial species selected from Rombutsia ilealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophilus, Enterococcus faecalis, Lactococcus piscium, Clostridium botulinum and Listeria monocytogenes is reduced relative to the abundance of the following anaerobic bacterial species before administration of CORM or a composition thereof. In particular, the abundance of Romboutsia ilealis is increased relative to the abundance of the anaerobic bacterial species before administration of CORM or a composition thereof in the intestinal microbiota of subjects with intestinal dysbiosis, especially in the case of a high-fat diet or drug treatment that induces dysbiosis.

[0092] In preferred embodiments, the abundance of the anaerobic bacterial species Akkermansia muciniphila is increased and the abundance of the anaerobic bacterial species Romboutsia ilealis is decreased relative to the abundance of these anaerobic bacterial species described below prior to administration of the CORM or composition thereof.

[0093] In some embodiments, the gut microbiota of a subject suffering from gut dysbiosis is restored or remodeled to a healthy phenotype.

[0094] In some embodiments, the frequency and / or severity of gut dysbiosis is reduced in a subject suffering from gut dysbiosis.

[0095] In some embodiments, the frequency and / or severity of nausea, diarrhea, constipation, bloating, gaz, and / or cramping is reduced in a subject suffering from intestinal dysbiosis.

[0096] In some embodiments, the CORM or composition thereof used according to the present invention increases the abundance of potentially beneficial bacterial species present in the intestinal microbiota, such as the abundance of one or more bacterial species selected from the group consisting of Akkermansia muciniphila, Alistipes fingoldii, Alternaria sabdariffa, Olsenella umbonata and Erwinia gingivalis, preferably Akkermansia muciniphila.

[0097] In some embodiments, the CORM or composition thereof used according to the present invention reduces the abundance of potentially harmful bacterial species present in the intestinal microbiota, such as the abundance of one or more bacterial species selected from the group consisting of Rombutsia ilealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophiles, Enterococcus faecalis, Lactococcus piscium, Carnobacterium divergens and Listeria monocytogenes, preferably Romboutsia ilealis.

[0098] In some embodiments, the CORM or composition thereof used according to the present invention promotes the maintenance of a healthy state of the subject's gut microbiota.

[0099] In some embodiments, CORMs or compositions thereof used according to the present invention help balance the intestinal microbiota.

[0100] In some embodiments, the CORM or composition thereof used according to the present invention promotes and / or maintains and / or restores good intestinal health.

[0101] In some embodiments, a CORM or composition thereof is administered before, during, and / or after a dietary regimen (eg, a high-fat diet) or a drug treatment (eg, antibiotic treatment) that is susceptible to impairing good intestinal health in a subject.

[0102] In a particular embodiment, the carbon monoxide releasing molecule is administered in a dose of 1 mg / kg to 25 mg / kg, preferably 2.5 mg / kg to 15 mg / kg.

[0103] The dosage of carbon monoxide releasing molecules (CORM) administered to a subject may vary depending on various factors such as the age, weight and / or sex of the subject, the diet that the subject has followed, follows and / or will follow, the proportion of fat in the subject's diet, the medication followed, such as the type of antibiotics used, and / or the degree of intestinal dysbiosis. Based on the knowledge in this field, a person skilled in the art would know how to determine the appropriate dosage to be administered to a subject based on these various factors.

[0104] In some embodiments, the carbon monoxide-releasing molecule or composition thereof is administered 1 to 7 times per week, preferably 3 to 6 times per week.

[0105] In some embodiments, carbon monoxide releasing molecules (CORMs) or compositions thereof are administered to a subject for one week, two weeks, three weeks, or longer, e.g., 1 month, 2 months, 3 months, or even longer if necessary, e.g., before, during, and / or after a high-fat diet or drug treatment that induces dysbiosis (e.g., antibiotic treatment).

[0106] In some embodiments, a carbon monoxide releasing molecule (CORM) or composition thereof is administered daily during a high fat diet or dysbiosis-inducing drug treatment (eg, antibiotic treatment).

[0107] In some embodiments, the carbon monoxide releasing molecule (CORM) or composition thereof is administered daily for at least three weeks, preferably one month, prior to a high fat diet or dysbiosis-inducing drug treatment (eg, antibiotic treatment).

[0108] In some embodiments, the carbon monoxide releasing molecule (CORM) or composition thereof is administered daily for at least three weeks, preferably one month, following a high fat diet or dysbiosis-inducing drug treatment (eg, antibiotic treatment).

[0109] In some embodiments, a carbon monoxide releasing molecule (CORM) or composition thereof is administered daily during a high-fat diet, and daily for at least three weeks, preferably one month, before and after a high-fat diet or dysbiosis-inducing drug treatment (e.g., antibiotic treatment).

[0110] In some embodiments, the CORM composition is a pharmaceutical composition or a nutraceutical composition.

[0111] In some embodiments, the CORM composition is prepared in a form suitable for oral or rectal use. The CORM composition can be in various forms, including, for example, capsules, tablets, powders, pills, dragees, granules, sachets, gels, pastes, syrups, emulsions, suspensions, solutions. Preferably, the CORM composition is in a form selected from capsules, tablets, powders, pills, dragees, granules, suppositories. More preferably, the CORM composition is in the form of capsules or pills.

[0112] In some embodiments, the CORM or composition thereof is encapsulated in a nanocapsule or contained in a micelle.

[0113] In some embodiments, the nutraceutical composition is a food supplement, such as a dietary supplement or a functional food. The functional food may be a beverage, a dairy-based product, or a fermented dairy-based product, such as yogurt.

[0114] One skilled in the art would know how to determine the appropriate concentration of the CORM to be included in the CORM composition depending on the dose of the CORM to be administered to a subject and the form of the composition.

[0115] In some embodiments, the CORM composition comprises 0.1% to 10% by weight, such as 1% to 5% w / w, such as 1% w / w or 3% w / w CORM relative to the total weight (w / w) of the composition.

[0116] In some embodiments, the prebiotic and / or probiotic is administered before, simultaneously with, and / or after a CORM or a composition thereof. The probiotics may be selected from one or more of the following bacterial species: Akkermansia muciniphila, Eubacterium limosum, Parasutterella secunda, Lactobacillus sporogenes, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus salivarius, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium lactis, Lactobacillus johnsonii, Lactobacillus paracasei, paracasei), preferably Akkermansia muciniphila, Eubacterium mucilaginosa, Parasutterella secunda. Prebiotics can be selected from one or more of the following ingredients: dietary fiber from fruits, vegetables, whole grains; fermented foods, such as kombucha, kefir, skyryogurt, fermented cottage cheese, vegetable brinedrink and / or fermented vegetables (such as kimchi). Preferably, prebiotics are selected from skim yogurt, vegetable brine, fermented vegetables.

[0117] Another object of the present invention relates to the use of CORM or a composition thereof for increasing the abundance of potentially beneficial bacterial species present in the intestinal microbiota of a subject, such as one or more bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Alternaria sabdariffa, Olsenella umbonata and Erwinia gingivalis, preferably Akkermansia muciniphila.

[0118] In some embodiments, the CORM or composition thereof is administered before, during, and / or after a dietary regimen (e.g., a high-fat diet) or a drug treatment (e.g., antibiotic treatment) that tends to reduce the abundance of potentially beneficial bacterial species present in the subject's gut microbiota.

[0119] Another object of the present invention relates to the use of CORM or a composition thereof for reducing the abundance of potentially harmful bacterial species present in the intestinal microbiota of a subject, such as one or more bacterial species selected from Rombutsia ilealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophilus, Enterococcus faecalis, Lactococcus piscium, Clostridium botulinum and Listeria monocytogenes, preferably Romboutsia ilealis.

[0120] In some embodiments, the CORM or composition thereof is administered before, during, and / or after a dietary regimen (e.g., a high-fat diet) or a drug treatment (e.g., antibiotic treatment) that tends to increase the abundance of potentially harmful bacterial species present in the subject's gut microbiota.

[0121] Another object of the present invention relates to the use of CORM or a composition thereof for promoting the maintenance of a healthy state of the intestinal microbiota of a subject.

[0122] In some embodiments, the CORM or composition thereof is administered before, during, and / or after a dietary regimen (e.g., a high-fat diet) or a drug treatment (e.g., antibiotic treatment) that compromises the subject's gut microbiota to maintain a healthy state.

[0123] Another object of the present invention relates to the use of CORM or a composition thereof for helping to balance the intestinal microbiota of a subject.

[0124] In some embodiments, the CORM or composition thereof is administered before, during, and / or after a dietary regimen (e.g., a high-fat diet) or a drug treatment (e.g., antibiotic treatment) that compromises the balance of the gut microbiota of a subject susceptible to a health state.

[0125] Another object of the present invention relates to the use of CORM or a composition thereof for promoting and / or maintaining and / or restoring good intestinal health in a subject.

[0126] Another object of the present invention relates to the use of CORM or a composition thereof for maintaining, restoring or remodeling the intestinal microbiota of a subject to a healthy phenotype.

[0127] In some embodiments, the CORM is selected from:

[0128] -Metal carbonyls, such as CORM-401 (CAS No.: 1001015-18-4), CORM-2 (CAS No.: 22594-69-0), CORM-3 (CAS No.: 475473-26-8), CORM-371, EBOR-CORM-1, ALF-186, ALF850, ALF021,

[0129] -Carbon monoxide myoglobin (CO-Mb), carboxyhemoglobin (CO-Hb) and polyethylene glycol-coupled hemoglobin (CO-MP4);

[0130] - metal-free CO-releasing compounds, such as CORM-A1 (CAS No. 17363-08-5); and / or

[0131] - Organic CO-releasing molecules, such as oxalic acid (CAS No. 144-62-7), cyclopentenone derivatives, cyclopentadienone-alkyne pairs and S-arylthioformates.

[0132] In some embodiments, the carbon monoxide releasing molecule is CORM-401 (CAS No: 1001015-18-4).

[0133] According to a preferred embodiment, the subject is a human.

[0134] In some embodiments, the CORM or composition thereof is administered before, during, and / or after a high fat diet or drug treatment (eg, antibiotic treatment).

[0135] The present invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted as limiting the scope of the present invention in any way.

[0136] Example

[0137] Embodiment 1: Blood carboxyhemoglobin (COHb) levels after oral administration of CORM-401

[0138] a) Materials and methods:

[0139] COHb (carbon monoxide bound to hemoglobin) levels were measured at different time points after oral administration of CORM-401 to two groups of 8-week-old male C57BL6 mice, which were fed with a standard diet (SD) or a high-fat diet (HFD) for 14 weeks (n=9-10 / group). CORM-401 was freshly prepared in phosphate buffered saline (PBS) and orally administered at a dose of 30 mg / kg=90 μmol / kg (n=6), and COHb in the blood was measured 1, 6, 24, and 48 hours later.

[0140] b) Results

[0141] like Figure 1 As shown, COHb levels in the blood were significantly elevated in a similar manner 1 hour after CORM-401 administration to SD-fed mice and HFD-fed mice, and gradually decreased to baseline levels over a 48-hour period. These data indicate that CO is released by CORM-401 and delivered to the blood circulation in a time-dependent manner. Thus, a reproducible increase in CO was achieved each time the compound was orally administered to mice over a 14-week treatment.

[0142] These results confirm the rationale for dosing CORM-401 three times weekly based on the kinetics of CO delivery by this compound in mice.

[0143] Embodiment 2: Levels of CO accumulation in different intestinal tissues after oral administration of CORM-401

[0144] a) Materials and methods

[0145] Eight-week-old male C57BL6 mice weighing approximately 25 g were treated with oral administration of CORM-401 (30 mg / kg) or PBS (control group) and then sacrificed 1, 3, 6, 24, and 48 hours later (n=6 / time point). Organs were collected to determine the level of CO accumulation in different intestinal tissues after both treatments. The body compartments where the intestinal microbiota is most likely to exist, such as the cecum, cecal contents, colon, and feces, were analyzed. CO accumulation was quantified using the CO-sensitive scavenger hemoCD1, as previously reported by the inventors, for example, in Mao et al., 2021

[10] , Minegishi et al., 2017

[11] . In brief, 50 mg of tissue was homogenized in 0.5 ml of PBS in the presence of hemoCD1 (2-10 μM) and sodium dithionite (1 mg). The tissue homogenate was first sonicated, then the sample was centrifuged, the supernatant was filtered, and then measured by UV-VIS absorption spectroscopy. The CO concentration was calculated based on the absorbance at 422 nm and 434 nm using the reported equation (Kitagishi et al.

[12] , 2010; Braud et al. 2018

[13] ).

[0146] b) Results

[0147] It can be observed that after CORM-401 treatment, CO in cecal tissue ( Figure 2 A) Cecal contents ( Figure 2 B), colon ( Figure 2 C) and excrement ( Figure 2 D) in a time-dependent manner. In all partitions, the maximum CO content was reached 6 hours after CORM-401 administration and returned to the basal level after 48 hours. Therefore, oral administration of CORM-401 to mice enables CO to be delivered and effectively reach tissues that directly interact with the intestinal microbiota.

[0148] Example 3 : Analysis of the effects of oral administration of CORM-401 on the abundance of anaerobic bacterial species in the gut microbiota

[0149] a) Materials and methods

[0150] 8-week-old male C57BL6 mice weighing about 25 g were fed with a standard diet (SD), a high-fat diet (HFD), or HFD+CORM-401 for 14 weeks (n=9-10 / group). CORM-401 was freshly prepared in phosphate buffered saline (PBS) and then orally administered to mice three times a week throughout the HFD regimen at a dose of 30 mg / kg=90 μmol / kg each time. Excreta from the three groups of mice were collected after 14 weeks of the HFD regimen for microbiota analysis before the mice were sacrificed.

[0151] Fecal sample DNA and RNA (n = 3 / group) were extracted and sequenced by shotgun metagenomic / metatranscriptomic techniques as previously described [9]. Briefly, a pre-extraction step combining mechanical, enzymatic, and chemical procedures was performed before extraction by the QIAsymphony PowerFecal Pro DNA kit (QIAGEN). The extracts were used for DNA / RNA library preparation, and pair-end sequences were determined using the NextSeq500 instrument (Illumina). Fastq files were analyzed by in-house software MetaMIC (v2.2.0). Data were filtered based on quality (phred score), and sequences were mapped to several custom databases, counted, and collected in separate Fastq files. Sequence counts were performed for each bacterial species, and the sequence counts were compared with environmental controls. The number of sequences for each species was used to determine their relative abundance in each group of mice.

[0152] b) Results

[0153] As shown in Table 3 below, the proportions of the 50 most common gut bacterial species present in SD-fed mice changed significantly after a 14-week HDF regimen with or without CORM-401. For example, the archetypal beneficial bacterium Akkermansia muciniphila was the second most abundant bacterium in SD-fed mice but did not appear as one of the most predominant bacteria in HFD-fed mice.

[0154] Interestingly, orally administered CORM-401 remodeled the microbiota profile and restored homeostasis of specific bacteria, whose relative abundance is presented in Table 3. This microbiota profile was confirmed when the relative abundance (number of DNA sequences) of each bacterial species was measured in the three groups.

[0155] As shown in Table 3, HFD-fed mice had significantly higher incidence of Lactococcus lactis, Enterococcus faecium, Streptococcus thermophilus, and the potentially harmful bacterial species Romboutsia ilealis, while Akkermansia muciniphila was significantly reduced compared to SD-fed mice. In particular, HFD mice treated with CORM-401 showed a gut bacterial profile characterized by Akkermansia muciniphila as one of the most dominant species, while the abundance of Alistipes fingoldii, Alternaria sabralina, Olsenella umbonata, and Erwinia gingivalis was significantly increased, while the abundance of Romboutsia ilealis was reduced.

[0156] These results clearly demonstrated that oral administration of CORM-401 remodeled the gut microbiota of HFD mice toward a healthy phenotype by increasing and / or decreasing the abundance of some specific anaerobic bacterial species present in the gut microbiota.

[0157] Table 3: Effect of oral administration of CORM-401 on the abundance and diversity of bacterial species present in the gut microbiota of HFD mice. Data are presented as the number of sequences (mean of 3 independent experiments per group).

[0158]

[0159]

[0160] References

[0161] [1]Passos et al (2017), Intestinal microbiota in digestive diseases", ArqGastroenterol, vol 54(3)

[0162] [2]https: / / www.inserm.fr / dossier / microbiote-intestinal-flore-intestinale /

[0163] [3] Suvoro et al., (2013), “Gut Microbiota, Probiotics, and Human Health”, Biosci Microbiota Food Health., 32(3):81–91

[0164] [4] Dave M. et al., (2012), “The human gut microbiome: current knowledge, challenges, and future directions”, Transl Res., 160:246-57.

[0165] [5] Vandenplas et al., (2015), “Probiotics: an update”, J Pediatr., 91(1):6-21.

[0166] [6] Devine et al., (2009), “Prospects for the development of probiotics and prebiotics for oral applications”, J Oral Microbiol., 1:10

[0167] [7] Cheng et al., (2021) 《Recent Advances on Carbon Monoxide Releasing Molecules for Antibacterial Applications》, ChemMedChem, 16, 3628–3634

[0168] [8] Herrmann WA., (1990), “100 Years of metal carbonyls: a serendipitous chemical discovery of major scientific and industrial impact”, J OrganometChem., 383:1-3

[0169] [9]Rodriguez C, et al., (2021), “Viral genomic, metagenomic and human transcriptomic characterization and prediction of the clinical forms of COVID-19”, PLoS Pathog., 17(3): e1009416.

[0170]

[10] Mao, Q., Kawaguchi, A.T., Mizobata, S., Motterlini, R., Foresti, R., & Kitagishi, H., (2021), “Sensitive quantification of carbon monoxide (CO) in vivo reveals a protective role of circulating hemoglobin in CO intoxication. Commun. Biol. 4, 425 - doi:10.1038 / s42003 - 021 - 01880 - 1.

[0171]

[11] Minegishi, S., Yumura, A., Miyoshi, H., Negi, S., Taketani, S., Motterlini, R., Foresti, R., Kano, K., & Kitagishi, H, (2017), “Detection and removal of endogenous carbon monoxide by selective and cell permeable hemoprotein - model complexes”. J. Am. Chem. Soc. 139, 5984 - 5991.

[0172]

[12] Kitagishi, H., Negi, S., Kiriyama, A., Honbo, A., Sugiura, Y., Kawaguchi, A.T., & Kano, K., (2010). “A Diatomic Molecule Receptor That Removes CO in a Living Organism. Angew. Chem. Int. Ed Engl.”, 49, 1312 - 1315.

[0173]

[13] Braud, L., Pini, M., Muchova, L., Manin, S., Kitagishi, H., Sawaki, D., Czibik, G., Ternacle, J., Derumeaux, G., Foresti, R., & Motterlini, R. (2018), “Carbon monoxide-induced metabolic switch in adipocytes improves insulin resistance in obese mice.”, JCI Insight 3, e123485

[0174]

[14] De Simone et al., “ThioCORMates: Tunable and Cost-Effective Carbon Monoxide-Releasing Molecules”, Chemistry, 2022 May 9. doi:10.1002 / chem.202201326。

Claims

1. A carbon monoxide releasing molecule (CORM) or a composition thereof for use in treating or preventing intestinal flora imbalance in a subject, wherein: The CORM or composition thereof is administered orally or rectally.

2. The CORM or composition thereof for use according to claim 1, wherein: CORM is selected from: -Metal carbonyls, such as CORM-401 (CAS No.: 1001015-18-4), CORM-2 (CAS No.: 22594-69-0), CORM-3 (CAS No.: 475473-26-8), CORM-371, EBOR-CORM-1, ALF-186, ALF850, ALF021, -Carbon monoxide myoglobin (CO-Mb), carboxyhemoglobin (CO-Hb) and polyethylene glycol-coupled hemoglobin (CO-MP4); - metal-free CO-releasing compounds, such as CORM-A1 (CAS No. 17363-08-5); and / or - Organic CO-releasing molecules, such as oxalic acid (CAS No. 144-62-7), cyclopentenone derivatives, cyclopentadienone-alkyne pairs and S-arylthioformates.

3. The CORM or composition thereof for use according to any one of claims 1 or 2, wherein: The carbon monoxide releasing molecule is CORM-401 (CAS No.: 1001015-18-4).

4. The CORM or composition thereof for use according to 1 to 3, wherein: The subject is a human.

5. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The intestinal flora imbalance is cecal and / or colonic flora imbalance.

6. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The CORM or composition thereof is administered before, during and / or after a dysbiosis-inducing dietary regimen, such as a high-fat diet, or a dysbiosis-inducing drug treatment, such as antibiotic treatment.

7. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The abundance of one or more bacterial species is increased and / or decreased relative to the abundance of the bacterial species prior to administration of the CORM or composition thereof.

8. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The abundance of the one or more bacterial species is normalized, or maintained at a normal abundance, relative to the abundance of the one or more bacterial species prior to administration of the CORM or composition thereof.

9. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The abundance of one or more anaerobic bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Alistipes shahii, Olsenella umbonata, Olsenella uli, Faecalibaculum rodentium, Lactobacillus murinus, Lactobacillus fermentum, Libanicoccus massiliensis, Lactobacillus salivarius, Lactobacillus agilis, Lactobacillus ruminis, Lactobacillus animalis, Parabacteroides goldsteinii, and Aerococcus viridans is increased relative to the abundance of these anaerobic bacterial species before administration of the CORM or a composition thereof.

10. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The abundance of the anaerobic bacterial species Akkermansia muciniphila is increased relative to its abundance prior to administration of the CORM or a composition thereof.

11. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: The abundance of one or more anaerobic bacterial species selected from Rombutsiailealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophiles, Enterococcus faecalis, Lactococcus piscium, Carnobacterium divergens and Listeria monocytogenes is reduced relative to the abundance of these anaerobic bacterial species before administration of the CORM or a composition thereof.

12. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The gut microbiota of subjects suffering from gut dysbiosis is restored or remodeled to a healthy phenotype.

13. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The frequency and / or severity of intestinal dysbiosis is reduced in subjects with intestinal dysbiosis.

14. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: The carbon monoxide releasing molecule is administered in a dosage of 1 mg / kg to 25 mg / kg, preferably 2.5 mg / kg to 15 mg / kg.

15. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: The carbon monoxide releasing molecule is administered 1 to 7 times per week, preferably 3 to 6 times per week.

16. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: CORM compositions are prepared in a form suitable for oral or rectal use.

17. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: The CORM composition is a pharmaceutical composition or a nutraceutical composition.

18. A CORM for use according to any one of the preceding claims, or a composition thereof, wherein: The CORM composition is selected from the group consisting of capsules, tablets, powders, pills, dragees, granules, sachets, gels, pastes, syrups, emulsions, suspensions, solutions.

19. A CORM or composition thereof for use according to any one of the preceding claims, wherein: The prebiotics and / or probiotics are administered before the CORM or a composition thereof, simultaneously with the CORM or a composition thereof, and / or after the CORM or a composition thereof.

20. Use of CORM or a composition thereof for increasing the abundance of potentially beneficial bacterial species present in the intestinal microbiota of a subject, wherein the potentially beneficial bacterial species is, for example, one or more bacterial species selected from Akkermansia muciniphila, Alistipes fingoldii, Alternaria sabdariffa, Olsenella umbonata and Erwinia gingivalis, preferably Akkermansia muciniphila.

21. Use of CORM or a composition thereof for reducing the abundance of potentially harmful bacterial species present in the intestinal microbiota of a subject, wherein the potentially harmful bacterial species is, for example, one or more bacterial species selected from Rombutsia ilealis, Lactococcus lactis, Enterococcus faecium, Streptococcus thermophilus, Enterococcus faecalis, Lactococcus piscium, Clostridium botulinum and Listeria monocytogenes, preferably Romboutsia ilealis.

22. Use of CORM or a composition thereof for promoting the maintenance of a healthy state of the intestinal microbiota of a subject.

23. Use of a CORM or a composition thereof to help balance the intestinal microbiota of a subject.

24. Use of a CORM or a composition thereof for promoting and / or maintaining and / or restoring good intestinal health in a subject.

25. Use of a CORM or a composition thereof for maintaining, restoring or remodeling the intestinal microbiota of a subject to a healthy phenotype.

26. The use according to any one of claims 20 to 25, wherein CORM is selected from: -Metal carbonyls, such as CORM-401 (CAS No.: 1001015-18-4), CORM-2 (CAS No.: 22594-69-0), CORM-3 (CAS No.: 475473-26-8), CORM-371, EBOR-CORM-1, ALF-186, ALF850, ALF021, -Carbon monoxide myoglobin (CO-Mb), carboxyhemoglobin (CO-Hb) and polyethylene glycol-coupled hemoglobin (CO-MP4); - metal-free CO-releasing compounds, such as CORM-A1 (CAS No. 17363-08-5); and / or - Organic CO-releasing molecules such as oxalic acid (CAS No. 144-62-7), cyclopentenone derivatives, cyclopentadienone-alkyne pairs and S-arylthioformates.

27. The use according to any one of claims 20 to 26, wherein The carbon monoxide releasing molecule is CORM-401 (CAS No.: 1001015-18-4).

28. The use according to any one of claims 20 to 27, wherein The subject is a human.

29. The use according to any one of claims 20 to 28, wherein The CORM or composition thereof is administered before, during and / or after a high fat diet or drug treatment, such as antibiotic treatment.

Images