Iron-Based Compositions for Use in Treating Gastrointestinal Inflammation in Subjects Treated for Iron Deficiency - Patent application

Abstract

The present invention relates to iron-based formulations and compositions thereof for use in a method for treating gastrointestinal inflammation by administering the formulation to a subject in the therapeutic or non-therapeutic treatment of iron deficiency, wherein the iron formulation is capable of reducing (or maintaining constant) the amount of pathogenic bacteria present in the intestinal microbiota of the treated subject, and preferably of increasing the variety of the intestinal microbiota of the treated subject.

Description

[Technical field]

[0001] The present invention relates to an iron-based preparation and a composition comprising the same for use in a method for treating gastrointestinal infection in a subject who has been treated for iron deficiency. The treatment method can reduce pathological side effects, such as gastrointestinal disorders, or their aggravation caused by administering an iron preparation in a subject who has been treated for iron deficiency, whether therapeutic or non-therapeutic. In particular, the iron preparation and a composition comprising the same can reduce the amount (or maintain constant or counteract the growth) of pathogenic bacteria present in the intestinal microflora of the subject, preferably belonging to the Proteobacteria phylum. Furthermore, the iron preparation and a composition comprising the same can advantageously increase the diversity of the intestinal microflora of the subject by shifting the balance towards species favorable to the organism, and therefore reduce the side effects caused.

[0002] It is known that after administration of an iron-based supplement selected from those currently available on the market, e.g. those based on Fe(II), a prevalence (or increase or elevation) of pathogenic bacteria (e.g. Proteobacteria) may occur in the (gastro)intestinal microflora of a treated subject taking the iron-based supplement.

[0003] Dysbiosis is a state of microbial imbalance caused by the overgrowth of so-called "bad" bacteria in the intestine, the microorganisms that cause its inflammation. The microbiome is a collection of symbiotic bacteria that coexist in our body, especially in the intestine, and plays a pivotal role in the control of immune tolerance, since it is closely related to the immune system. When the balance of the intestinal microbiome is altered, it is called dysbiosis, a disorder that can also be caused by malnutrition and is associated with chronic inflammatory diseases, obesity, tumors and colitis.

[0004] The increase in gastrointestinal pathogens following iron supplementation is particularly evident in geographic regions or subjects where the presence of gastrointestinal pathogens is endemic (e.g., developing countries and / or malnourished subjects). The prevalence of gastrointestinal pathogenic bacteria increases the risk of gastrointestinal infection and diseases and / or disorders associated with gastrointestinal infection, such as intestinal inflammation, gastrointestinal ulcers, chronic gastritis, gastric atrophy, intestinal diverticulum, diarrhea, vomiting, nausea, and gastrointestinal pain.

[0005] Gastrointestinal infection or disorders also reduce the absorption of iron administered by iron-based supplements.

[0006] Furthermore, subjects with iron deficiency, even if mild, generally exhibit an imbalance in the gut microbiota, particularly a reduced diversity of the gastrointestinal microbiota associated with a reduction in beneficial bacteria known to be producers of short-chain fatty acids, which means that subjects with iron deficiency (but not necessarily anemic) are more susceptible to gastrointestinal infections than subjects not affected by iron deficiency.

[0007] Document WO2019 / 025922 relates to a composition comprising an iron(III) salt, a phospholipid and a sucrester for treating iron deficiency.

[0008] The article XP036744615 by Abbati G et al. relates to the use of an iron-based composition called Sucrosomial® Iron for treating gastrointestinal disorders.

[0009] Article XP055665686 by Susana Gomez-Ramirez et al. relates to the use of an iron-based composition called Sucrosomial® Iron to treat gastrointestinal disorders.

[0010] Article XP055665690 by Angela Fabiano et al. relates to a study on the gastrointestinal absorption of an iron-based composition generally referred to as Sucrosomial® Iron, but does not disclose the treatment of gastrointestinal inflammation.

[0011] The article XP085262682 by Angela Fabiano et al. relates generally to studies on absorption in in vivo and in vitro models, but does not disclose treatment of gastrointestinal inflammation.

[0012] Das Nupur et al.'s paper XP055855831 relates generally to the gut microbiota and its regulation.

[0013] To date, there are many iron-based supplements on the market that can cause gastrointestinal side effects such as diarrhea, nausea, vomiting and abdominal pain.

[0014] Furthermore, excess iron in the digestive lumen can lead to the proliferation of pathogenic bacteria in the intestine that negatively affect the basic gut microbiome, which can lead to intestinal dysbiosis and increase the risk of gastrointestinal infections.

[0015] In view of the above, the technical problem addressed and solved by the present invention is to provide an iron supplement composition which, in addition to providing iron, prevents or reduces gastrointestinal infections or disorders, such as pathological side effects associated with the administration of common iron-based supplements.

[0016] In other words, the technical problem that the present invention addresses and resolves is to provide an iron supplementation composition which, in addition to supplementing iron, does not cause and / or prevents an increase of pathogenic bacteria (e.g., Proteobacteria) in the (gastro)intestinal microbiota and / or is effective in restoring balance and proper diversity of the (gastro)intestinal microbiota and, as a result, prevents or reduces gastrointestinal infections or disorders, e.g. pathological side effects associated with common iron-based supplements.

[0017] Furthermore, there is a need for iron supplement compositions that do not exacerbate the endemic presence of pathogenic bacteria (e.g., Proteobacteria) in the gastrointestinal microbiota, such as in subjects with diseases that lead to disorders of the gut microbiota (IBD, celiac disease, etc.) or in subjects living with inadequate nutrition or poor hygiene.

[0018] Restoring the appropriate diversity of the (gastro)intestinal microbiota mainly, but not exclusively, consists in reducing or avoiding the increase in pathogenic bacteria (Proteobacteria) and increasing bacteria capable of producing short-chain fatty acids.

[0019] After intense research and development activity, the present applicant addresses and solves the above reported technical problems by providing what is described and claimed: a solid form of iron preparation comprising an iron(III) salt, a phospholipid and sucroster for the treatment (therapeutic or non-therapeutic) of iron deficiency (briefly, the iron preparation according to the present invention or sucrosomal iron).

[0020] The iron preparation according to the invention (Sucrosomial® Iron) has been shown to increase the diversity of the intestinal microbiota to a greater extent than a comparative composition, for example containing ferrous sulfate. In particular, the iron preparation according to the invention has been shown to have a positive effect on the reduction of pathogenic bacteria belonging to the phylum Proteobacteria and on the increase of bacteria capable of producing short-chain fatty acids.

[0021] These and other objects, which will become apparent from the following detailed description, can be achieved by the solid form iron preparations and related compositions of the present invention as a result of the technical features set out in the specification and described in the appended claims. [Brief description of the drawings]

[0022] [Figure 1] Figure 1A-D shows body weight (A), liver iron concentration (B), serum iron (C) and transferrin saturation (D) in three groups of mice (ID, FS and IS). All data represent the mean ± SEM of 10 mice (***P<0.005). [Diagram 2] Figures 2A-D show red blood cell count (A), hemoglobin (B), hematocrit (C) and mean corpuscular volume (D). All data represent the mean ± SEM of 10 mice (*P<0.05, ***P<0.005). [Diagram 3]Figure 3: Measurement of microbiota composition by beta diversity in fecal samples collected from each mouse after 2 weeks with a deficient diet (A or baseline) and after 2 weeks with the iron-containing diet under analysis (B or FS_B, ID_B, SI_B). [Figure 4] Figure 4A-B are measurements of microbial diversity in a single sample; Figure 4A shows the evenness, richness and Shannon index for each group of animals; Figure 4B shows the change in Shannon index and richness in each mouse after 2 weeks with a specific diet (B) relative to baseline (A). [Diagram 5] 5A and 5B depict the differences in microbial composition at the phylum (Proteobacteria and Bacteroides) level in mice fed sucrosomial iron (IS) or ferrous sulfate (FS): directional changes occurring in the microbiome of each mouse after 2 weeks with the specific diets indicated (B) versus baseline (A). [Figure 6] 6A-6G depict differences in microbial composition at the amplicon single variant (Lachnospiracea spp. (asv33; asv24; asv72), Rikenellacea RC9 (asv30; asv151), Faecalibaculum (asv90; asv18), Corydextrin (asv124), Oscillibacter (asv163), Parasterella (asv70) and Lactobacillus (johnsonii_asv2)) level in mice fed sucrosomial iron (IS) or ferrous sulfate (FS): directional changes occurring in the microbiome of each mouse after 2 weeks with the specific diets indicated (B) versus baseline (A). Summary of the Invention

[0023] The Applicant has discovered that when a subject undergoes iron-based treatment, regardless of the reasons that led to the initiation of such treatment (which may relate to iron deficiency due to a pathological condition or the need to restore iron levels due to, for example, intense sports activity), two cases occur very frequently depending on the physical and health condition of the subject.

[0024] In the first case, one may treat a subject who, in addition to already suffering from a gastrointestinal pathological disorder or condition, also suffers from iron deficiency, which is why the subject in question is undergoing an iron-based treatment. In this situation, a subject taking a commercially available iron supplement, for example based on iron(II), will experience a significant worsening of the gastrointestinal pathological condition due to the intake of an inappropriate form of iron, iron salt or iron ion.

[0025] In the second case, one may deal with subjects who, despite being in good health, need to restore their iron levels for reasons that are not of pathological origin or nature, such as sportsmen / women. Also in this situation, subjects taking commercial iron supplements, for example based on iron(II), may develop gastrointestinal pathological conditions due to the intake of inappropriate forms of iron, iron salts or iron ions.

[0026] The applicant has discovered that the formulations and compositions containing same contain iron or iron salts or iron ions in a special and suitable form that does not cause changes or alterations in the intestinal microflora that favor the spread of harmful pathogenic species. This effect is achieved regardless of whether the formulations of the invention are administered in the presence of a gastrointestinal infection or dysbiosis.

[0027] It is an object of the present invention to provide a solid form of iron preparation for use in a method for treating gastrointestinal inflammation (i.e. a method for reducing and / or preventing pathological side effects) in a subject undergoing therapeutic or non-therapeutic treatment for iron deficiency by administering a solid form of iron preparation of the present invention.

[0028] It is a further object of the present invention to provide an iron composition for use in a method for the treatment of gastrointestinal inflammation (i.e. a method for reducing and / or preventing pathological side effects) in a subject undergoing therapeutic or non-therapeutic treatment for iron deficiency, said composition comprising an iron preparation in solid form and at least one additive and / or excipient.

[0029] It is a further object of the present invention to provide a method for treating gastrointestinal disorders (i.e., a method for reducing and / or preventing pathological side effects) by administering to a subject a therapeutically effective amount of an iron preparation or composition administered to treat iron deficiency. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] The subject of the present invention is an iron preparation in solid form for use in a method for treating gastrointestinal inflammation (and / or diseases and / or symptoms associated therewith) caused or not caused by an infection in a subject who has undergone a therapeutic (pathological subjects) or non-therapeutic (healthy subjects) treatment for iron deficiency, by administering an iron preparation according to the invention, said iron preparation reducing or maintaining constant the amount of pathogenic bacteria present in the intestinal microflora of the treated subject, said preparation (for short, the preparation according to the invention) being: (a) iron as an iron salt or iron complex or iron oxide, preferably an iron salt; (b) at least one phospholipid; (c) Sucrester or sucrose ester A formulation comprising or consisting of:

[0031] Another object of the present invention is a solid iron(III) formulation for use in a method for treating gastrointestinal inflammation in a subject who has previously undergone therapeutic treatment for iron deficiency by administering the iron(II) compound or salt to the subject, the iron(III) formulation reduces the amount of pathogenic bacteria belonging to the Proteobacteria phylum present in the intestinal microbiome of the subject, the iron(III) formulation comprising or consisting of (a) an iron(III) salt or complex, (b) at least one phospholipid, (c) a sucrose or sucrose ester.Preferably, the subject is a subject with gastrointestinal inflammation and anemia, which has previously been treated with a separate and independent treatment based on an iron(II) compound or salt, such as iron(II) sulfate, but has been unsuccessful in view of not showing signs of healing or worsening gastrointestinal inflammation, such as IBD.Preferably, the treatment method is for treating a subject with gastrointestinal inflammation, such as IBD, and anemia; preferably, the gastrointestinal inflammation is of recurrent type.

[0032] Advantageously, in addition to reducing or maintaining constant the amount of pathogenic bacteria present in the intestinal microflora of the treated subject, the solid form of the iron preparation also increases the diversity of the intestinal microflora of the treated subject.

[0033] The solid form of the iron preparation according to the invention, which comprises phospholipids, sucrose and vegetable starch, is commercially referred to as sucrosomial® iron (sucrosomial® is a trademark registered on behalf of Pharmanutra SpA and Alesco Srl).

[0034] Further, an object of the present invention is a composition for use in a method for treating gastrointestinal inflammation and / or diseases and / or conditions associated therewith (according to any of the embodiments described herein) in the therapeutic or non-therapeutic treatment of iron deficiency by administering the composition to a subject, said composition (briefly, the composition of the present invention) comprising or consisting of an iron preparation of the present invention according to any one of the embodiments described and at least one pharmaceutical or food grade additive and / or excipient.

[0035] A method for treating gastrointestinal inflammation, preferably inflammation due to infection, provides for administration of a therapeutically effective amount of an iron preparation or composition of the invention to treat iron deficiency and simultaneously prevent the onset of gastrointestinal disorders, particularly gastrointestinal infection, or worsening if already present, and / or alleviate gastrointestinal disorders.

[0036] The gastrointestinal inflammation or a disease or condition associated therewith may be selected from the group consisting of intestinal inflammation, gastrointestinal ulcers, chronic gastritis, gastric atrophy, intestinal diverticulum, diarrhea, vomiting, gastrointestinal pain, nausea, constipation.

[0037] Examples of pathogenic bacteria present in the intestinal microflora belonging to the phylum Proteobacteria, the amount of which is reduced or maintained by administering an iron preparation or composition according to the invention, are pathogenic bacteria selected from the group comprising or consisting of Escherichia coli, Helicobacter pylori, Salmonella typhi, Vibrio cholerae, Streptococci.

[0038] The iron preparations or compositions of the invention are advantageously for use in the treatment of gastrointestinal inflammation in subjects having or at risk of a gastrointestinal infection or disorder, such as subjects with inadequate nutritional intake, subjects living in inadequate hygiene conditions, subjects with celiac disease, subjects with IBD, subjects who have undergone bariatric surgery, subjects with gastritis, subjects with gastric atrophy, subjects with gastroesophageal reflux (or gastroesophageal reflux disease, GERD).

[0039] In the context of the present invention, the expression "inadequate" is used to indicate less than the minimum level considered necessary for a subject to be considered healthy by the person skilled in the art.

[0040] The iron preparations or compositions of the invention may be administered to a subject to treat iron deficiency, such as non-pathological iron deficiency, anemia (e.g., subjects with lower hemoglobin concentrations compared to limits established by the World Health Organization (WHO)), sideropenia, iron deficiency in pregnant or lactating women or during the menstrual cycle, iron deficiency in the elderly, or iron deficiency in those with a compromised immune system.

[0041] The iron preparations or compositions of the invention may be administered to healthy subjects for non-therapeutic treatment to result in an increase in blood iron (e.g. sideremia, transferrin, ferritin levels) to improve physical performance (e.g. sportsmen and women) or mental performance (e.g. for learning reasons).

[0042] In addition to (a) an iron salt or complex, (b) at least one phospholipid and (c) a sclerotoxin, the formulation for use in the present invention may further comprise (d) a gelatinized or pre-gelatinized plant-derived starch, preferably rice starch (Oryza sativa) or maize starch, more preferably pre-gelatinized rice starch.

[0043] The (a) iron present in the formulation of the invention (the formulation comprising (a), (b), (c) and optionally (d)) may be an iron(II) or iron(III) salt or complex, preferably iron(III), selected from organic or inorganic iron salts such as chloride, sulfate, pyrophosphate, citrate, bisglycinate, fumarate, gluconate, ascorbate, polymaltosate, etc.; preferably, the iron consists of iron pyrophosphate (iron(III) pyrophosphate, Fe4(P2O7)3).

[0044] In the context of the present invention, the term "iron complex" is used to denote a coordination complex consisting of one or more iron atoms and one or more atoms, ions or molecules which at least partially surround it without forming ionic or covalent bonds (ligands).

[0045] For example, a formulation for use according to the invention comprises or consists of (a) iron pyrophosphate, (b) a phospholipid, preferably lecithin, (c) sucrestor, and optionally (d) pregelatinized rice starch.

[0046] The (b) at least one phospholipid present in the formulation of the invention (the formulation comprising (a), (b), (c) and optionally (d)) may be a phosphoglyceride selected from the group consisting of 1,2-diacyl-phospholipids, 1-alkyl-2-acyl-phospholipids and 1-alkenyl-2-acyl-phospholipids, preferably 1,2-diacyl-phospholipids, such as phosphatidylserine, phosphatidylcholine or lecithin, phosphatidylethanolamine, phosphatidylinositol or phosphatidic acid.

[0047] According to a preferred embodiment, (b) the at least one phospholipid in the formulation for use according to the invention (comprising (a), (b), (c) and optionally (d)) is a lecithin, preferably a lecithin (e.g. E322) selected from the group consisting of sunflower lecithin, corn lecithin, soy lecithin (e.g. allergen-free).

[0048] For example, a formulation for use according to the invention comprises or consists of (a) iron pyrophosphate, (b) lecithin, preferably sunflower lecithin, (c) sucrestor, and optionally (d) pregelatinized rice starch.

[0049] The (c) sucrose starch present in the formulation of the present invention (a formulation comprising (a), (b), (c) and, optionally, (d)) may be sucrose starch E473, preferably sucrose starch (e.g., E473) comprising at least two sucrose monoesters with fatty acids of plant origin, preferably fatty acids consisting of stearic acid and / or palmitic acid, in an amount of 70-90% by weight relative to the total weight of the sucrose starch, and, optionally, (d) gelatinized or pregelatinized starch of plant origin, preferably pregelatinized rice starch. According to a preferred embodiment, the formulation for use according to the present invention comprises (a) iron pyrophosphate, (b) sunflower lecithin (e.g., E322), (c) sucrose starch E473, preferably sucrose starch (e.g., E473) comprising at least one sucrose monoester of stearic acid and one sucrose monoester of palmitic acid, in an amount of 70-90% by weight relative to the total weight of the sucrose starch, and (d) pregelatinized rice starch.

[0050] According to one embodiment, a formulation for use according to the invention (comprising (a), (b), (c) and optionally (d)) comprises (c) sucrose and (b) a phospholipid, preferably lecithin, in a weight ratio of [(c):(b)] comprised between 50:1 and 10:1 (e.g. about 40:1, 35:1, 30:1, 25:1, 20:1, 17:1 or 15:1), preferably between 35:1 and 15:1.

[0051] According to one example, the solid form iron preparation of the present invention (comprising (a), (b), (c) and optionally (d)) comprises 1% to 40% (e.g., 5%, 10%, 15%, 20%, 25%, 30% or 35%), preferably 5% to 20% (e.g., about 10% to 12%, equivalent to about 44% to 45% weight / weight iron pyrophosphate) of elemental iron (or metallic iron or iron(III)) by weight relative to the total weight of the preparation.

[0052] According to a further example, the solid form iron preparation of the present invention (comprising (a), (b), (c) and optionally (d)) comprises the following weight percentages of the components relative to the total weight of the preparation: (a) 1% to 40% (e.g., 5%, 10%, 15%, 20%, 25%, 30% or 35%), preferably 5% to 20% elemental iron (or metallic iron or iron(III)) (e.g., about 10% to 12%, equivalent to about 44% to 45% weight / weight iron pyrophosphate); (b) 0.05% to 15% (e.g., 0.5%, 0.1%, 1%, 2%, 2.5%, 3%, 4%, 6%, 8% or 10%), preferably 0.1% to 2% (e.g., about 0.5±0.1% or 1.0±0.1%) of a phospholipid, preferably a lecithin (e.g., solid sunflower lecithin (E322)); (c) 1% to 50% (e.g., 3%, 5%, 7%, 10%, 12%, 15%, 17%, 20%, 25%, 30%, or 40%), preferably 10% to 25%, e.g., about 17% to 18% ± 1%) of a sclerode; and optionally, (d) 10% to 85%, for example 15%, 20%, 30%, 35%, 40%, 50% or 60%, preferably 25% to 50% (for example about 37 to 38±1%) of a starch of plant origin, preferably pregelatinized rice starch.

[0053] The term lecithin is used to describe a class of chemical compounds present in animal and plant tissues, especially egg yolks. Lecithin is a natural emulsifier and has antioxidant properties.

[0054] From a chemical point of view, lecithin consists of phosphatidylcholine ((R)-1-oleoyl-2-palmitoyl-phosphatidylcholine) or contains at least one phospholipid, preferably phosphatidylcholine, as the main component. Phosphatidylcholine is a phosphoglyceride in which phosphatidic acid is esterified with choline. Phosphatidic acid formally represents the simplest phosphoglyceride, produced by esterifying glycerol at the 1- and 2-positions with fatty acids and at the 3-position with orthophosphoric acid.

[0055] The (b) phosphatidylcholine or lecithin contained in the formulation of the present invention (comprising (a), (c) and optionally (d)) is preferably selected from sunflower lecithin, corn lecithin and soybean lecithin, more preferably lecithin in solid form (e.g. powder or granule), such as solid lecithin E322, even more preferably solid sunflower lecithin (e.g. E322), such as allergen-free solid sunflower lecithin E322 (preferably in the form of powder or granule). The term "allergen-free" means that it does not contain allergen residues. The term "E322" indicates that lecithin is a food additive (emulsifier) ​​permitted by European law and regulated by Italian Ministerial Decree No. DM 1996.

[0056] Directive 2008 / 84 / EC of 27 August 2008 (published in the Official Journal of the European Union No L253) lays down the purity criteria that lecithin must fulfil in order to be considered to meet the food quality criteria (Lecithin E322): acetone insolubles (i.e. active substances) min. 60%; water max. 2%; acidity number max. 35; peroxide number max. 10; toluene insolubles (i.e. impurities) max. 0.3%.

[0057] The (b) phosphatidylcholine or lecithin (preferably sunflower lecithin E322) in the form of powder or granules contained in the formulation of the present invention may, for example, have a moisture content of 1.5% to 4.5% (e.g. 2%, 2.5%, 3.5% or 4%) by weight relative to the weight of the lecithin; and / or a glucose content of 20% to 60% (e.g. 30%, 40%, 45% or 50%) by weight relative to the weight of the lecithin. An example of (b) sunflower lecithin E322 (powder or granule) that may be used in connection with the present invention may have the following composition (chemical / physical analysis, % weight / weight): 40% to 50% sunflower lecithin, 40% to 50% (e.g. about 42%) carbohydrates, 6% to 10% protein, 3% to 8% ash, 2% to 5% moisture and 0.5% to 1.5% of another flow agent.

[0058] According to one embodiment, the (b) lecithin contained in the formulation of the present invention (comprising (a), (c), and optionally (d)) does not comprise or consist of degraded or hydrolyzed lecithin, and does not comprise or consist of enzymatically degraded or hydrolyzed lecithin (i.e., enzymatically degraded or hydrolyzed).

[0059] The (c) sucrose starch (or synonymous with sucrose starch, sucrose ester or carbohydrate fatty acid ester) contained in one embodiment of the formulation of the present invention (comprising (a), (b) and optionally (d)) may be sucrose starch E473, preferably sucrose starch (E473) containing 50% to 95% (e.g. 60%, 70%, 80%, 85% or 90%, preferably 70% to 90%) by weight of a monoester obtained by esterifying sucrose with one or more fatty acids of plant origin, preferably stearic acid and / or palmitic acid, based on the total weight of sucrose starch. The term "E473" indicates that sucrose starch is a food additive (emulsifier) ​​permitted by European law and regulated by Italian Ministerial Decree No. DM 1996.

[0060] Sucresters are generally obtained by esterifying fatty acids or by transesterifying the methyl esters of fatty acids with carbohydrates (also called sugars); the carbohydrates used are sucrose (monosaccharides) and polysaccharides (hence the name "sucrose esters of fatty acids"). The chemical / physical properties of these compounds depend on the number and type of esterified fatty acids. Sucresters are essentially emulsifiers and are added to compositions to increase the stability of the hydrophilic and lipophilic phases.

[0061] An example of (c) sucrose star (E473) that may be used in the context of the present invention is a sucrose star having a hydrophilic-lipophilic balance (HLB) value comprised between 14 and 18 (e.g. 15, 16 or 17) and / or the following composition (% weight / weight): a total ester content of at least 90%, of which at least 70% are monoesters obtained by esterifying sucrose with one or more fatty acids of vegetable origin, preferably stearic acid and / or palmitic acid; a free fatty acid content (e.g. oleic acid) of not more than 3%; a free sucrose content of not more than 2%; a moisture content of not more than 4%; an acid value of not more than 5. An example of a commercially available (c) sucrose star is sucrose ester SP70 manufactured by Chimab SpA (Italia).

[0062] According to one embodiment, the (c) sucrose contained in the formulation of the present invention (containing (a), (b) and optionally (d)) does not contain or consist of a polyglycerol fatty acid ester.

[0063] The (d) pregelatinized rice starch optionally included in the formulation of the present invention (comprising (a), (b) and (c) and optionally (e)) may have, for example, the following chemical / physical properties: moisture content of 7% or less; protein content of 1% or less; ash content of 1% or less; pH (solution 10%) of 5.5 to 7.5; density of 0.40 to 0.48 g / cm 3 a minimum starch content of 97%; and a fat content of 0.1% or less. An example of a commercially available pregelatinized rice starch that can be used in connection with the present invention is AX-FG-P manufactured by Reire Srl (Italia).

[0064] The components (a), (b), (c) and (d) described in the present invention may be prepared according to techniques and equipment known to those skilled in the art and are commercially available.

[0065] The composition of the invention may be a pharmaceutical composition, a composition for a medical device (Medical Device Regulation (EU) 2017 / 745 (MDR)), a dietary supplement and / or a food for special medical purposes (FSMP).

[0066] The at least one pharmaceutical or food grade additive and / or excipient contained in the composition of the present invention together with the solid form of the iron preparation of the present invention consists of substances having no therapeutic activity suitable for pharmaceutical or food use selected from auxiliary substances known to those skilled in the art, such as diluents, solvents, solubilizers, thickeners, sweeteners, flavor enhancers, dyes, lubricants, surfactants, antimicrobial agents, antioxidants, preservatives, anti-caking agents, excipients, antistatic agents, pH stabilizing buffers and mixtures thereof. Non-limiting examples of such substances are hydroxymethylcellulose, dye E171, vegetarian magnesium stearate, magnesium salts of fatty acids, silicon dioxide, crystalline cellulose.

[0067] In addition to the iron preparations and additives / additives of the present invention, the compositions of the present invention may further comprise (e) at least one vitamin, preferably selected from the group comprising or consisting of (ei) vitamin C, (e-ii) at least one B vitamin, preferably vitamin B6 and / or B9 and / or B12, (e-iii) D vitamins, preferably vitamin D3, (e-iv) vitamin E, and mixtures thereof; preferably (ei) vitamin C and / or (e-ii) at least one B vitamin.

[0068] (e) At least one vitamin may be present in the composition in an amount between 500% RDA and 300% RDA, such as 100% RDA, 150% RDA, 200% RDA or 250% RDA (RDA: Recommended Dietary Allowance).

[0069] The (e) at least one vitamin contained in the composition of the present invention may be a sucrosomial (registered trademark) vitamin, which is a solid form formulation comprising (e) at least one vitamin, (b) a phospholipid (e.g., lecithin), (c) a sucrose starch, and optionally (d) a vegetable starch (e.g., pregelatinized rice starch), similar to any of the embodiments described for the solid form sucrosomial (registered trademark) iron preparation of the present invention.

[0070] Examples of compositions according to the present invention comprising sucrosomial iron and at least one sucrosomial vitamin are: iron and at least one vitamin selected from vitamin C, vitamin D3, vitamin B (preferably B12), vitamin E, and mixtures thereof; iron and vitamin C; iron and vitamin D3; iron, vitamin C and vitamin D3; iron, vitamin C and vitamin B12; iron, vitamin D3 and vitamin B12; iron, vitamin C, vitamin D3 and vitamin B12.

[0071] The compositions of the invention according to any one of the described embodiments are preferably formulated in a solid form, such as a neat solid form or an orally dissolvable solid form (or orally soluble, i.e., dissolves in the oral cavity) or a water-dispersible solid form (i.e., dissolves in a water-based liquid), such as a powder, granules, microgranules, flakes, tablets or capsules.

[0072] Alternatively, the compositions of the invention according to any one of the embodiments described may be in liquid form, such as a solution, dispersion or suspension of a solid in a liquid, or in semi-solid form, such as a cream, gel or softgel.

[0073] The formulations or compositions of the present invention are preferably formulated for oral administration, eg, as an intact solid form, an orally dissolvable or water-dispersible solid form.

[0074] The composition of the present invention in tablet form may have a weight comprised between 200mg and 2000mg, for example a hard tablet between 800mg and 1000mg. The tablet may be coated or filmed with one or more coating layers or films capable of passing through the gastric barrier. The coating may comprise beeswax or a sugar-based solution.

[0075] The composition of the invention in the form of a capsule may have a weight comprised between 100 mg and 1500 mg, more preferably about 800 mg; said capsule may be of hard or soft gelatin or softgel.

[0076] The iron preparation of the invention (sucrosomial® iron) according to any one of the described embodiments may be prepared according to the first method described in patent document WO2014 / 009806A1, page 7, line 1 to page 8, line 20, which is incorporated herein by reference, or according to the second method described in patent document WO2014 / 009806A1, page 8, line 22 to page 10, line 21, which is incorporated herein by reference.

[0077] For example, a method for producing an iron formulation of the present invention may include the steps of: (1) producing an iron salt or complex in the form of a powder or granules; (2) mixing said iron salt or complex with a phospholipid, preferably a lecithin (e.g., sunflower lecithin) and at least one sclerotoxin, to obtain a mixture of step 2 or a formulation of the present invention, advantageously wherein the lecithin is not hydrolyzed or enzymatically hydrolyzed lecithin; (3) optionally, mixing the mixture of step 2 with a plant extract. (4) optionally, after step 2 or step 3, by sieving the resulting product through a sieve having a nominal mesh size of 150 μm to 1400 μm (e.g., 180, 212, 250, 300, 355, 425, 500, 600, 710, 850, 1000, or 1180 μm), preferably 600 μm to 850 μm, e.g., 710 μm (25 US mesh).

[0078] Preferably, the mixing in steps 2 and / or 3 is carried out at room temperature (15° C. to 30° C., preferably about 25° C.) for 10 minutes to 120 minutes, preferably 15 minutes to 60 minutes, for example, about 30 minutes.

[0079] According to a preferred embodiment, the mixing in steps 2 and / or 3 is carried out in the absence of a solvent (dry mixing) and / or the formulation process of the iron formulation of the present invention does not include a spray drying step.

[0080] Furthermore, step 2 of mixing iron with phospholipids and at least one sclerester may be divided into two substeps, the first substep providing for mixing iron with phospholipids and the second substep providing for mixing the mixture obtained in the first substep with at least one sclerester.

[0081] The process for producing the iron preparation of the present invention is carried out using equipment known to those skilled in the art, for example by mixing components (a), (b), (c) and optionally (d) according to the weight percentages defined for the present invention.

[0082] The at least one sucrosomial® vitamin optionally included in the compositions of the invention along with the iron preparation of the invention may be prepared according to the methods for preparing the iron preparations of the invention described herein, substituting at least one vitamin for iron.

[0083] In the context of the present invention, the term "subject" is used to refer to mammals (animals and humans), preferably human subjects, males and females, e.g. subjects of pediatric age, adults, elderly subjects, subjects with diseases and subjects normally active in sports activities.

[0084] The term "therapeutically effective amount" is used to indicate an amount of an iron preparation that elicits a biological or medical response in a tissue, system or subject as determined and defined by one of skill in the art.

[0085] Unless otherwise specified, a statement of a formulation or composition that contains an amount of an ingredient "ranging from x to y" is used to indicate that the ingredient may be present in the formulation or composition in any amount between the endpoints of that range, even if not specified.

[0086] Unless otherwise specified, the content of an ingredient in a formulation or composition refers to the weight percentage of that ingredient relative to the total weight of the formulation or composition.

[0087] Unless otherwise specified, a statement that a formulation or composition "comprises" one or more ingredients means that other ingredients (other than the one or more specifically named) may be present, and a statement that a formulation or composition "consists of" a given ingredient means that the presence of other ingredients is excluded.

[0088] An embodiment (FRn) of the present invention is shown below.

[0089] FR1 An iron preparation for use in a method for treating gastrointestinal inflammation or a disease or condition related thereto in a subject who has been treated therapeutically or non-therapeutically for iron deficiency by administering the iron preparation to the subject, comprising: The iron preparation reduces the amount of pathogenic bacteria belonging to the phylum Proteobacteria present in the intestinal microbiota of the treated subject, and The iron preparation, (a) an iron salt or complex, (b) at least one phospholipid; (c) Sucrester or sucrose ester A formulation comprising or consisting of:

[0090] FR2 A formulation for use according to FR1, wherein the method of treatment is for reducing gastrointestinal inflammation; preferably, the inflammation is due to an infection.

[0091] FR3 A formulation for use according to FR1 or FR2, wherein the pathogenic bacteria belonging to the phylum Proteobacteria comprise or are selected from the group consisting of Escherichia coli, Helicobacter pylori, Salmonella typhi, Vibrio cholerae, and Streptococcus.

[0092] FR4 A formulation for use according to any one of FR1 to FR3, wherein the iron formulation increases or modifies the type of gut microbiota in the treated subject.

[0093] FR5. A formulation for use according to any one of FR1 to FR4, wherein the formulation further comprises (d) gelatinized or pregelatinized starch of vegetable origin; preferably rice starch or corn starch; more preferably pregelatinized rice starch.

[0094] FR6 (b) A formulation for use according to any one of FR1 to FR5, wherein the at least one phospholipid consists of a lecithin; preferably a lecithin (E322) selected from the group consisting of sunflower lecithin, corn lecithin, soybean lecithin and mixtures thereof.

[0095] FR7 (c) A formulation for use according to any one of FR1 to FR6, wherein the sucrose ester or sucrose ester is Sucrester E473; preferably a Sucrester (e.g. E473) comprising at least two sucrose monoesters having 30 to 95% by weight, preferably 50 to 85% by weight, of fatty acids of vegetable origin, preferably stearic acid and / or palmitic acid, based on the total weight of the Sucrester.

[0096] The FR8 formulation (a) iron pyrophosphate, (b) sunflower lecithin; (c) sucrose or sucrose esters; and (d) Pregelatinized rice starch comprising or consisting of Preferably, the formulation for use according to any one of FR1 to FR7, wherein the weight ratio between (c) sucrose ester or sucrose ester and (b) phospholipid or lecithin, [(c):(b)], is comprised between 50:1 and 10:1, preferably between 40:1 and 10:1, more preferably between 30:1 and 15:1.

[0097] FR9 The formulation for use according to any one of FR1 to FR8, wherein the gastrointestinal infection and / or disease or condition associated therewith is selected from the group consisting of intestinal inflammation, gastrointestinal ulcer, chronic gastritis, gastric atrophy, intestinal diverticulum, diarrhea, vomiting, gastrointestinal pain, nausea, constipation.

[0098] FR10 The subject has or is at risk for a gastrointestinal infection; Preferably, the formulation for use according to any one of FR1 to FR9, wherein the subject having or at risk of having a gastrointestinal infection is selected from the group consisting of subjects with inadequate nutritional intake, subjects living in inadequate hygiene conditions, subjects with celiac disease, subjects with IBD, subjects who have undergone bariatric surgery, subjects with gastritis, subjects with atrophic gastritis, subjects with gastroesophageal reflux disease (GERD).

[0099] FR11 composition, A formulation according to any one of the preceding clauses, and At least one pharmaceutical or food grade additive and / or excipient A composition for use according to any one of FR1 to FR10, comprising or consisting of: EXAMPLES

[0100] The following reports are some examples of solid form iron preparations according to the present invention. [Table 1] [Table 2] [Table 3] [Table 4]

[0101] Experimental Part Part 1

[0102] 1. Purpose of the study The effect of sucrosomial® iron, also called Sideral® RM (iron(III) according to the invention according to Table A.1) on the intestinal microbiome of mice is determined and compared with the effect of ferrous sulfate FeSO4 (iron(II) for comparison). The comparison was made with the same iron element in the salts used (iron in ionic form as Fe(III) on the one hand and Fe(II) on the other).

[0103] 2. Method Thirty 4-week-old C57BL / 6 male mice were fed an iron-deficient diet (based on AIN-93G rodent chow, approximately 1 mg / kg iron). All mice were housed in the same environment (QIMR Berghofer Animal Facility) to minimize inherent variation in the microbiome between individual mice.

[0104] At 6 weeks of age (i.e., 2 weeks on the iron-deficient diet), feces were collected for microbiome analysis. This was a control sample for each mouse, representing the basal microbiome of that particular animal when luminally iron-deficient. At this time point (6 weeks of age, 2 weeks on the deficient diet), the mice were presumed to have reduced iron stores but were not anemic, thus avoiding the potential further effects of anemia on the microbiome that could complicate the analysis.

[0105] After fecal collection at 6 weeks of age, mice were switched from an iron-deficient diet to a diet containing 50 mg / kg of iron, such as ferrous sulfate (iron(II) sulfate) or sucrosomial® iron (10 mice per group).

[0106] The remaining 10 mice served as a control group that remained on the deficient diet.

[0107] Fecal samples were then collected from each mouse at 8 weeks of age (i.e., 2nd week on the iron-containing diet), the mice were sacrificed, and tissues were harvested for analysis. Liver and serum samples were collected from each animal after euthanasia and stored at -80°C. Hematological parameters were assessed using a Coulter Ac·T diff Hematology Analyzer (Beckman Coulter, Gladesville, NSW, Australia) immediately after euthanasia. Serum iron parameters were assessed using the Iron / TIBC Reagent Kit (Pointe Scientific, Canton, MI), and iron levels in tissues were determined as previously described (Frazer, DM, et al. Cell Mol Gastroenterol Hepatol 3, 410-421 (2017)). For the analysis of iron and hematological parameters, statistical differences between groups were calculated using one-way ANOVA followed by Tukey post-hoc test for groups with equal variances and Games-Howell post-hoc test for groups with unequal variances. Significant differences in variances were identified using Levene's test. DNA was collected from fecal samples and microbial profiles (QIMR Berghofer Medical Research Institute) were determined.

[0108] Group of mice under test: ID: control group that remained on iron-deficient diet (approximately 1 mg / kg iron). FS: Group fed diet containing 50 mg / kg of iron in the form of ferrous sulfate (FS is an abbreviation for ferrous sulfate) - Fe(II). SI: group fed diet containing 50 mg / kg iron in the form of sucrosomial® iron (SI is the abbreviation for sucrosomial iron).

[0109] 3.Results 3.1 Iron status in mice fed sucrosomial® iron or ferrous sulfate-based diets.

[0110] Prior to determining the effects on the microbiome, iron status was analyzed in the three analyzed groups of rats identified in section 2 (ID, FS, and SI).

[0111] After sacrifice at 8 weeks of age, the liver and serum iron levels, as well as hematological parameters, of the three groups of mice analyzed (ID, FS, SI) were examined. All data in Figure 1A-D represent the mean ± SEM of 10 mice per group (*P<0.05, ***P<0.005).

[0112] There were no significant differences in body weights among the three groups (ID, FS and SI; Fig. 1A). There were no significant differences in liver iron concentrations, serum iron levels, transferrin saturation, red blood cell counts, hemoglobin, hematocrit or mean corpuscular volume between the groups treated with sucrosomial® iron and ferrous sulfate (Figs. 1B-D, 2A-D). However, in each case, the means of the iron-deficient groups (ID, control) were significantly lower than those with an iron-containing diet (FS and SI) (Figs. 1B-D, 2A-D). These animals (ID, control) had been kept on an iron-deficient diet for 4 weeks, so it is not unexpected that they would show signs of iron deficiency.

[0113] Having determined that the iron status was similar to groups treated with iron (sucrosomial® iron (SI) or ferrous sulfate (FS)), changes in the microbiome of these animals were examined.

[0114] 3.2 Overall microbial composition – beta diversity Beta diversity compares the overall microbial composition between samples, i.e., what identified microbial profiles different samples have. Principal coordinate analysis (PCoA) classifies samples based on their overall microbial community in a two-dimensional space. Each symbol in Figure 3 represents a sample in a format that indicates the diet and time point (i.e., baseline, or after an additional 2 weeks on a particular diet). The reference sample is a sample taken from each mouse after the first 2 weeks (baseline) of the iron-deficient diet. The distance between each sample indicates how similar / dissimilar the sample is to the other samples (i.e., the further away, the less similar). The data show that samples taken from mice fed either an iron-rich diet (iron sulfate (FS) or sucrosomial iron (IS)) were largely grouped together and could be distinguished from samples taken from mice fed an iron-deficient diet (ID). This indicates that the iron-containing diet was able to alter the overall microbial diversity. Both cage effects (i.e., animals housed together have more similar microbiomes) and mouse effects (i.e., the microbiome was observed to change over time in the same mouse, even on an iron-deficient diet) were observed. These effects are not uncommon for mice and were corrected for in subsequent analyses.

[0115] 3.3 Overall microbial composition – alpha diversity Alpha diversity measures the diversity of microorganisms within a single sample. Several parameters are used to measure the diversity of a sample. Richness refers to the number of different microbial sequences detected and is an estimate of the number of bacterial strains in a sample. Evenness indicates how evenly bacteria are distributed within a sample, i.e., whether there is a dominant species. The Shannon index is a combination of richness and evenness for each sample. An increase in the Shannon index is generally accepted as good for health. In this sample, diversity changed significantly using unpaired statistics (ANOVA). The reference sample (baseline) showed the lowest richness, followed by the iron-deficient group (ID) and the iron-rich diets (FS and SI). The Shannon index was lower in mice fed ferrous sulfate and higher in mice fed sucrosomial iron (Figure 4A). This indicates that the overall gut microbial health improved with the sucrosomial iron (SI)-containing diet versus the iron-deficient diet (ID) and the ferrous sulfate (FS)-containing diet. We then performed a mixed-effects regression analysis with "mouse" and "cage" as random effects, "diet" as independent variable, and "time" as fixed effect, with the diversity indices (Shannon, richness, Chao 1, distinct richness, evenness index) of changes due to individual heterogeneity of the gut microbiota as outcomes (Figure 4B). A significant increase in the Shannon index was observed by switching to a sucrosomial iron-containing diet alone, again suggesting that this diet produces a healthy gut microbiome.

[0116] 3.4 Changes at the phylum level While diversity indices measure the overall health of the microbiome, it is useful to test the types of bacteria present. Many sequences obtained by 16S sequencing cannot yet be assigned to specific bacterial species, but many of the amplicons can be grouped into taxonomic classifications that provide information about ongoing changes. Taxonomic classification is based on a taxonomic classification hierarchy that includes higher ranks that are very general (e.g., phyla) and lower ranks that are very specific (e.g., genera). For all the following analyses, P values ​​of taxon richness as outcomes were obtained from mixed-effects regression models with “mouse” and “cage” as effects, “feed” as independent variable, and “time” as fixed effect. P values ​​were adjusted using Bonferroni correction to avoid false positives. P values ​​less than 0.05 were considered significant.

[0117] At the phylum level, Proteobacteria were significantly reduced in the sucrosomial iron (SI)-containing diet but not in the iron-deficient (ID) or ferrous sulfate (FS)-containing diet (Figure 5). Proteobacteria is a phylum that includes many pathogens such as Escherichia, Enterobacter and Salmonella. Previous studies have shown that iron supplements can increase the prevalence of gastrointestinal pathogens in areas where they are endemic. Although these pathogens were not present in our animals, the reduction of Proteobacteria in the sucrosomial iron (SI) group, accompanied by a protective intestinal effect of this iron supplement, suggests that sucrosomial iron is a better option compared to other iron supplements in areas where intestinal pathogens are endemic.

[0118] Members of the phylum Bacteroidota were significantly reduced in mice fed a diet containing ferrous sulfate (FS) but remained unchanged in a diet containing sucrosomial iron (SI) (Figure 5). A significant increase was observed in an iron-deficient diet. The reasons for these changes are unclear, but Bacteroidota is one of the major phyla that inhabit the gut and are involved in the production of short-chain fatty acids. A reduction in Bacteroidota has been associated with obesity.

[0119] 3.5 Changes at the amplicon single variant (ASV) level ASVs represent the actual sequenced amplicons. Because microbiomes are complex, many have not been fully characterized at the species level. In the groups fed the sucrosomial iron (SI) diet, various ASVs were increased, including Lachnospiraceae spp., Rikenellaceae RC9, Faecalibaculum, Colidextribacter and Oscillibacter (Figures 6A-6E). These bacterial groups are generally associated with a healthy microbiome due to their ability to produce short-chain fatty acids.

[0120] Faecalibaculum has been shown to have a protective effect against intestinal tumors. This is interesting because studies in rodents have shown that ferrous sulfate (FS) can promote the development of intestinal cancer.

[0121] In contrast, bacteria from the genus Parasutterella were reduced in the sucrosomial iron (IS) group (Figure 6F). Parasutterella species are also associated with irritable bowel syndrome and chronic intestinal inflammation, although bacterial strains belonging to this genus are considered normal members of the gut microbiome.

[0122] Lactobacillus was reduced at the genus level only in animals treated with sucrosomial iron (SI) (Figure 6G). Although Lactobacillus species are considered beneficial, this reduction should not be considered a negative effect of sucrosomial iron, considering that this genus is one of the few groups of organisms that do not require iron for survival. Therefore, this result likely represents a proportional increase in Lactobacillus species during the first two weeks of the iron-deficient diet, when the growth of other bacteria that require iron is suppressed. Switching to a sucrosomial iron (SI)-containing diet allows the growth of other bacterial species, enhancing the overall diversity and health of the gut microbiome. The resulting increase in competing species reduces Lactobacillus to more normal levels.

[0123] The observation that there was no similar reduction in Lactobacilli in animals receiving a ferrous sulfate (FS)-based diet may reflect a lack of diversity in this group, with Lactobacilli levels remaining high due to fewer competing species.

[0124] A lack of diversity is a sign of an unhealthy microbiome.

[0125] 4. Conclusion Overall, the changes in the gut microbiome induced by a sucrosomial iron (IS)-containing diet are beneficial, increasing microbial diversity compared to a ferrous sulfate (FS)-containing or iron-deficient diet. Increased microbial diversity is generally believed to produce a healthier microbiome. More generally, increased diversity is associated with a reduction in pathogenic bacteria belonging to the phylum Proteobacteria.

[0126] Thus, a reduction in the phylum Proteobacteria was observed in the Sucrosomial® Iron (SI) diet. The reduction in Proteobacteria, a phylum that includes many pathogenic bacterial strains, with the Sucrosomial® Iron-based diet is potentially beneficial as iron-based supplements have been shown to increase the risk of intestinal infections and their effectiveness is reduced in areas where enteric pathogens are endemic.

[0127] Sucrosomial® Iron (IS) also promoted the growth of beneficial bacteria known for producing short-chain fatty acids.

[0128] Based on the above, the results suggest that sucrosomial® iron has more beneficial effects on the gut microbiome compared to ferrous sulfate (FS) in a mouse model.

[0129] Part 2 Study Design

[0130] (Introduction) Iron is an essential component present in hemoglobin in red blood cells and myoglobin in muscles, and is also essential for the function of many cellular activities. However, iron can be toxic as it catalyzes the production of reactive oxygen species, and therefore, iron homeostasis is well controlled in mammals, especially in the duodenum, where iron is mainly absorbed. The gut microbiota represents the totality of microorganisms present in the gastrointestinal tract (GIT). The gut microbiota includes all commensal and pathogenic bacteria that reside in the gastrointestinal GIT. The gut microbiota plays an important role in the absorption of minerals and nutrients, the synthesis of enzymes, vitamins and amino acids, and the production of short-chain fatty acids (SCFAs). However, there is a potential risk that these mechanisms may be compromised as a result of changes in the microbial composition, also known as dysbiosis.

[0131] The gut microbiota encounters various concentrations of unabsorbed iron, derived from the diet, supplements administered orally, or iron-based drugs. Iron is essential for bacteria, as it functions as a cofactor for iron-containing proteins involved in mechanisms present in redox reactions, metabolic models, and the electron transport chain, and the iron status in the host and the availability of iron present in the feed can alter this microbial medium.

[0132] Oral administration of iron can increase the proliferation and viability of enteropathogenic bacteria, leading to diarrhea and intestinal inflammation, whereas iron deficiency also alters the microbial composition.

[0133] One way to address iron deficiency is to supplement with iron-based products; however, this approach is not always effective and can cause significant side effects.

[0134] To overcome the incidence of side effects due to supplementation with iron-based products by the oral route, new iron-containing formulations for oral use are needed that are effective in protecting iron and increasing intestinal absorption so as to reduce dosage and side effects.

[0135] The composition of the present invention, which comprises or consists of (a) an iron salt or complex (iron(III) pyrophosphate), (b) at least one phospholipid, and (c) a sucrose ester or sucrose ester, has the advantage of having iron(III) pyrophosphate protected by the sucrose and phospholipid matrix. The composition of the present invention is mostly in the form of vesicle-like structures that overwhelm traditional iron absorption models (in vitro tests). Due to this gastrointestinal behavior, the composition of the present invention is well tolerated and has high bioavailability compared to traditional iron salts such as ferrous sulfate(II).

[0136] Since oral administration of iron can alter the composition of the gut microbiota due to the ability of microorganisms to metabolize iron by increasing the proliferation of pathogenic bacteria, the aim of this experimental part is to investigate the alterations in the fecal microbiome in iron-deficient subjects administered iron(III) pyrophosphate and iron(II) sulfate according to the present invention.

[0137] (Main purpose of the test) To evaluate changes in fecal microbiota during supplementation with two different iron formulations: Sucrosomial® Iron (a composition of the present invention containing iron(III) pyrophosphate) versus iron(II) sulfate.

[0138] (Secondary objective of the study) Fecal calprotectin concentrations will be assessed.

[0139] (Primary endpoint of the study) The first goal of this study is to compare the composition of the fecal microbiota in terms of different bacterial populations, their abundance and biodiversity, and to evaluate whether different iron preparations may result in different microbiota compositions. The baseline microbiota taken before treatment will be compared with three different time points during and after treatment.

[0140] (Secondary endpoint of the study) The secondary goal of the study will focus on evaluating possible changes in fecal calprotectin and gaining a better understanding of its correlation with alterations in the fecal microbiota.

[0141] (Population under test) The study recruited 20 subjects, including both anemic and nonanemic but iron-deficient healthy subjects. Subjects reported having gastrointestinal disorders or in need of iron-based nutritional therapy. Recruitment was by routine visits when iron deficiency or a very low daily iron dose was detected.

[0142] (placement) General data were collected during recruitment: sex, age, spontaneous habits, BMI, bowel movement frequency, and dietary habits. In clinical practice, iron supplementation is proposed to patients with anemia or clinical or biochemical signs of iron deficiency.

[0143] Subjects were assigned to either Treatment A (Sucrosomial® Iron) or Treatment B (Ferrous Sulfate) Groups 1 or 2. A 6 ml blood sample was taken prior to administration of the iron-based product and initiation of treatment.

[0144] After 14 days, blood samples are taken for determination of secondary outcomes, and the procedure is repeated with secondary administration of iron.

[0145] (treatment) Recruited subjects were iron deficient and were treated with Sucrosomial® iron 30 mg or ferrous sulfate 105 mg. The study was an open-label, non-randomized, design in which all subjects received Sucrosomial® iron and ferrous sulfate compositions (open-label, non-randomized, controlled entry crossover design).

[0146] Treatment Group: Group 1: i) 2 capsules / day of Sucrosomial® iron for 14 days; ii) 1 capsule / day of ferrous sulfate for 14 days. Group 2: i) 1 capsule / day of ferrous sulfate for 14 days; ii) 2 capsules of Sucrosomial® iron for 14 days. All subjects will have a 4-day washout period between the two treatments (Treatment A, Sucrosomial® iron; Treatment B, ferrous sulfate). Subjects will be crossed over from one treatment to the other on an informed and open-label basis.

[0147] The procedure proceeds as follows: - 2 weeks of treatment with ferrous sulfate or Sucrosomial® iron - 4-day washout - 2 weeks of treatment with ferrous sulfate or Sucrosomial® iron

[0148] Fecal samples for microbiota analysis are collected as follows: - T0 = before treatment begins - T1=14 days - T2=18 days - T3=32 days Within 2 hours of collection, the samples are sent to a laboratory where they are stored at -80°C to maintain the quality of DNA and RNA. Genomic DNA is extracted from the fecal samples using a QIAmp PowerFecal Pro DNA Kit (QIAGEN, Hilden, Germany). DNA concentration is calculated by measuring OD260nm, and DNA purity is estimated by determining the ratios of OD260 / OD280 and OD260 / OD230 using a BioPhotometer D30 spectrophotometer (Eppendorf).

[0149] (Metagenomic analysis) The DNA genome extracted from the fecal samples is subjected to metagenomic analysis by sequencing the genome encoding 165 rRNA bacteria. Next, NGS (Next Generation Sequencing) and data analysis are performed using Novogene. Different regions of the gene, namely V3-V4, are amplified using Phusion® High-Fidelity PCR Master Mix (New England BioLabs, USA). PCR products are purified with QIAGEN Gel Extraction Kit (QIAGEN), libraries are generated with NEBNext® ultraTM DNA Library Prep Kit for illumina, and quantified with Qubit and qPCR. Sequencing amplicons are performed on the HiSeq illumina platform. The obtained sequences are grouped into OUTs by 97% similarity of DNA sequences considered to collect homologous species. Sequence analysis is performed using Uparse software. Phylogenetic relationships between all OUTs are determined using the multiple comparison program MUSCLE. Alpha and beta diversity are calculated from the results.

[0150] (Inclusion criteria) - Target age group: 18-70 - Iron deficiency: low iron levels in the blood and serum ferritin levels below 25 μg / l - BMI between 18.5 and 24.9 kg / m 2 - Symptoms of iron deficiency include extreme fatigue, weakness, pale skin, high heart rate or shortness of breath or difficulty breathing, headache, and irritated or red tongue. - Inadequate dietary intake of iron, normal calprotectin levels, as assessed by a nutritionist - Written informed consent

[0151] (exclusion criteria) Subjects who meet at least one of the following criteria will be considered ineligible: - Subjects taking drugs that alter the composition of the microbiota, such as PPIs (proton pump inhibitors), antibiotics, or long-term use of synbiotics or probiotics, during the 4 weeks prior to the start of the study - Subjects diagnosed with IBD, chronic hepatitis or pancreatitis, recent infection with C. difficile, Salmonella, Campylobacter, Shigella, or aggressive infection with E. coli - Subjects who have received a blood transfusion within the past 4 months - Pregnant subjects - Subjects who have been breastfeeding for at least 6 months prior to the start of the study - Smokers who smoke more than one cigarette per week

[0152] Data collection Fecal calprotectin. The gut microbiota is dominated by four main phyla: Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria.

[0153] (statistical analysis) A p-value of less than 0.05 was considered statistically significant. Analyses are performed using SPSS (version 21, IBM Corp, Armonk, NY).

[0154] A preferred embodiment (FRn) of the present invention is shown below.

[0155] FRP1. An iron(III) preparation for use in a method for treating gastrointestinal inflammation in a subject who has previously received therapeutic treatment for iron deficiency by administering to said subject an iron(II) compound or salt, comprising: - the iron(III) preparation reduces the amount of pathogenic bacteria belonging to the phylum Proteobacteria present in the intestinal microflora of the treated subject, and - the iron(III) preparation (a) an iron(III) salt or complex, (b) at least one phospholipid; (c) Sucrester or sucrose ester A formulation comprising or consisting of:

[0156] FRP2 An iron(III) preparation for use as described in FRP1, wherein the subject to be treated is a subject with gastrointestinal inflammation and is anemic and has previously been treated unsuccessfully with an iron(II) compound or salt based treatment.

[0157] FRP3 An iron(III) preparation for use according to FRP1 or FRP2, wherein the method of treatment is for treating a subject with gastrointestinal inflammation and anemia, preferably wherein the gastrointestinal inflammation is of the recurrent type.

[0158] FRP4. A formulation for use according to any one of FRP1 to 3, wherein the method of treatment is for reducing gastrointestinal inflammation; preferably, the inflammation is due to an infection.

[0159] FRP5 The formulation for use according to any one of FRP1 to 4, wherein the pathogenic bacteria belonging to the phylum Proteobacteria include or are selected from the group consisting of Escherichia coli, Helicobacter pylori, Salmonella typhi, Vibrio cholerae, and Streptococcus.

[0160] FRP6 A formulation for use according to any one of FRP1 to 5, wherein the iron(III) formulation increases or alters the variety of intestinal microbiota in the treated subject.

[0161] FRP7 A formulation for use according to any one of FRP1 to 6, wherein the iron(III) formulation further comprises (d) gelatinized or pregelatinized starch of plant origin; preferably rice starch or corn starch; more preferably pregelatinized rice starch.

[0162] FRP8 (b) A formulation for use according to any one of FRP1 to 7, wherein at least one phospholipid consists of lecithin; preferably a lecithin (E322) selected from the group consisting of sunflower lecithin, corn lecithin, soybean lecithin and mixtures thereof.

[0163] FRP9 (c) A formulation for use according to any one of FRP1 to 8, wherein the sucrose ester or sucrose ester consists of sucrose ester E473; preferably a sucrose ester (e.g. E473) comprising at least two sucrose monoesters having 30 to 95% by weight, preferably 50 to 85% by weight, of fatty acids of vegetable origin, preferably fatty acids consisting of stearic acid and / or palmitic acid, relative to the total weight of the sucrose ester.

[0164] FRP10 iron(III) preparation, (a) iron(III) pyrophosphate, (b) sunflower lecithin; (c) sucrose or sucrose esters; and (d) Pregelatinized rice starch comprising or consisting of Preferably, the formulation for use according to any one of FRP1 to 9 has a weight ratio of [(c):(b)] between (c) sucrose ester or sucrose ester and (b) phospholipid or lecithin of 50:1 to 10:1, preferably 40:1 to 10:1, more preferably 30:1 to 15:1.

[0165] FRP11 The formulation for use according to any one of FRP1 to 10, wherein the gastrointestinal infection is selected from the group consisting of intestinal inflammation, gastrointestinal ulcers, chronic gastritis, gastric atrophy, intestinal diverticulum, diarrhea, vomiting, gastrointestinal pain, nausea, constipation.

[0166] FRP12 The subject has or is at risk for a gastrointestinal infection; Preferably, the subject having or at risk of having a gastrointestinal infection is selected from the group consisting of subjects with inadequate nutritional intake, subjects living in inadequate hygiene conditions, subjects with celiac disease, subjects with IBD, subjects who have undergone bariatric surgery, subjects with gastritis, subjects with atrophic gastritis, subjects with gastroesophageal reflux disease (GERD).

[0167] The FRP13 composition is A formulation according to any one of the preceding clauses, and At least one pharmaceutical or food grade additive and / or excipient A composition for use according to any one of FRP1 to 12, comprising or consisting of:

Claims

1. An iron(III) formulation for treating gastrointestinal inflammation in a subject who has previously been treated therapeutically for iron deficiency by administering to the subject an iron(II) compound or salt, comprising: The iron(III) preparation is (a) an iron(III) salt or complex, (b) at least one phospholipid that is a lecithin; (c) Sucrester or sucrose ester A formulation comprising or consisting of:

2. 2. The iron(III) formulation of claim 1, wherein the subject to be treated is a subject with gastrointestinal inflammation and is anemic and has been previously treated unsuccessfully with an iron(II) compound or salt-based treatment.

3. The formulation according to claim 1 or 2, wherein the method of treatment is for treating a subject with gastrointestinal inflammation and anemia, preferably the gastrointestinal inflammation is of the recurrent type.

4. The formulation according to any one of claims 1 to 3, wherein the method of treatment is for reducing gastrointestinal inflammation; preferably, the inflammation is due to an infection.

5. A formulation described in any one of claims 1 to 4, wherein the inflammation is caused by a pathogenic bacterium belonging to the phylum Proteobacteria.

6. The formulation of claim 5, wherein the pathogenic bacteria is selected from the group including or consisting of Escherichia coli, Helicobacter pylori, Salmonella typhi, Vibrio cholerae and Streptococcus.

7. The method according to any one of claims 1 to 6, wherein the iron(III) preparation increases the diversity of intestinal microflora in a treated subject.

8. 8. The iron(III) formulation according to any one of claims 1 to 7, wherein the iron(III) formulation further comprises (d) gelatinized or pregelatinized starch of vegetable origin; preferably rice starch or corn starch; more preferably pregelatinized rice starch.

9. 9. The formulation according to any one of claims 1 to 8, wherein (b) the at least one phospholipid consists of a lecithin (E322) selected from the group consisting of sunflower lecithin, corn lecithin, soybean lecithin and mixtures thereof.

10. (c) A formulation according to any one of claims 1 to 9, in which the sucrose ester or sucrose monoester comprises sucrose monoester E473, which comprises 30 to 95% by weight, preferably 50 to 85% by weight, based on the total weight of the sucrose monoester, of at least one sucrose monoester having fatty acids of vegetable origin, preferably stearic acid and / or palmitic acid.

11. Iron(III) preparations, (a) iron(III) pyrophosphate, (b) sunflower lecithin; (c) sucrose or sucrose esters; and (d) Pregelatinized rice starch comprising or consisting of A formulation according to any one of claims 1 to 10, wherein the weight ratio between (c) the sucrose ester or sucrose ester and (b) the phospholipid or lecithin, [(c):(b)], is preferably comprised between 50:1 and 10:1, preferably between 40:1 and 10:1, more preferably between 30:1 and 15:

1.

12. The formulation according to any one of claims 1 to 11, wherein the gastrointestinal infection is selected from the group consisting of intestinal inflammation, gastrointestinal ulcers, chronic gastritis, gastric atrophy, intestinal diverticulum, diarrhea, vomiting, gastrointestinal pain, nausea and constipation.

13. the subject has or is at risk for a gastrointestinal infection; Preferably, the subject having or at risk of having a gastrointestinal infection is selected from the group consisting of subjects with inadequate nutritional intake, subjects living in inadequate hygiene conditions, subjects with celiac disease, subjects with IBD, subjects who have undergone bariatric surgery, subjects with gastritis, subjects with atrophic gastritis, subjects with gastroesophageal reflux disease (GERD), A formulation according to any one of claims 1 to 12.

14. The composition comprises: A formulation according to any one of claims 1 to 13, and At least one pharmaceutical or food grade additive and / or excipient A composition comprising or consisting of:

15. An iron(III) preparation for reducing the amount of pathogenic bacteria belonging to the phylum Proteobacteria, preferably Escherichia coli, Helicobacter pylori, Salmonella typhi, Vibrio cholerae and Streptococcus, present in the intestinal microflora of a subject, comprising: The iron(III) preparation is (a) an iron(III) salt or complex, (b) at least one phospholipid that is a lecithin; (c) Sucrester or sucrose ester A formulation comprising or consisting of:

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