WO2025224128A1 - Méthodes de traitement de maladies associées à une surcharge en fer par administration locale d'hepcidine dans l'intestin - Google Patents

Méthodes de traitement de maladies associées à une surcharge en fer par administration locale d'hepcidine dans l'intestin

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Publication number
WO2025224128A1
WO2025224128A1 PCT/EP2025/060984 EP2025060984W WO2025224128A1 WO 2025224128 A1 WO2025224128 A1 WO 2025224128A1 EP 2025060984 W EP2025060984 W EP 2025060984W WO 2025224128 A1 WO2025224128 A1 WO 2025224128A1
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Prior art keywords
hepcidin
iron
subject
gut
mice
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Pending
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PCT/EP2025/060984
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English (en)
Inventor
Carole Peyssonnaux
Marion FALABREGUE
Philippe Langella
Luis BERMUDEZ- HUMARAN
Anne AUCOUTURIER
Sophie Vaulont
Candice AURRAND
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Centre National de la Recherche Scientifique CNRS
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Cite
Institut des Sciences et Industries du Vivant et de lEnvironnement AgroParisTech
Universite Paris Saclay
Institut National de Recherche pour lAgriculture lAlimentation et lEnvironnement
Original Assignee
Centre National de la Recherche Scientifique CNRS
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Cite
Institut des Sciences et Industries du Vivant et de lEnvironnement AgroParisTech
Universite Paris Saclay
Institut National de Recherche pour lAgriculture lAlimentation et lEnvironnement
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Application filed by Centre National de la Recherche Scientifique CNRS, Institut National de la Sante et de la Recherche Medicale INSERM, Universite Paris Cite, Institut des Sciences et Industries du Vivant et de lEnvironnement AgroParisTech, Universite Paris Saclay, Institut National de Recherche pour lAgriculture lAlimentation et lEnvironnement filed Critical Centre National de la Recherche Scientifique CNRS
Publication of WO2025224128A1 publication Critical patent/WO2025224128A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid

Definitions

  • the present invention relates to a method for preventing or treating iron overload associated diseases such as P-thalassemia and haemochromatosis by administration hepcidin locally in the gut.
  • Iron is an essential nutrient for vital biological process including hemoglobin synthesis, DNA replication and mitochondrial function.
  • perturbations of iron homeostasis still affect more than 25% of the world’s population 1,2 .
  • iron deficiency is the most common nutritional disorder in the world and is associated with anemia 1 .
  • hereditary hemochromatosis characterized by iron overload is the genetic disorders with the higher prevalence in the world.
  • the iron overload in hemochromatosis is due to a genetic mutation inducing a defect of systemic hepcidin and for consequences an increase of iron absorption 3 .
  • Hepcidin is an hyposideremic hormone produced by the liver 4 .
  • Intestine is the only organ enables to absorb iron, especially in the proximal part: duodenum.
  • the only source of iron for organisms is the food (l-2mg iron absorbed each day).
  • Non-heme iron dietary exists more largely as ferric form (Fe 3+ ) and is insoluble and no bio- available for organisms.
  • enterocyte cells reduce ferric iron in ferrous iron (Fe 2+ ) via the ferric reductase enzyme (DcytB) present at the brush border.
  • Divalent Metal Transporter 1 (DMT1) is the ferrous iron importer localized at the apical side.
  • iron can be stored in ferritin (FTL/FTH) or exported in the circulation via the basolateral exporter ferroportin (FPN).
  • FPN basolateral exporter ferroportin
  • Systemic hepcidin regulates iron absorption by inhibiting iron export (FPN).
  • numerous in vitro studies on intestinal cells line showed that hepcidin had no effect on FPN 1 1 l4 .
  • transgenic mice overexpressing the peptide specifically in this tissue (transgenic hepcidin). They next generate probiotics producing hepcidin to confirm the role of intestinal hepcidin on iron absorption (exogen hepcidin).
  • a first object of the invention relates to a method for preventing or treating an iron overload associated disease in a subject in need thereof, comprising administering locally in the gut of the subject a therapeutically effective amount of hepcidin.
  • a second object of the invention relates to a recombinant food-grade bacterium comprising a gene coding for hepcidin for use for preventing or treating an iron overload associated disease in a subject in need thereof, said recombinant food-grade bacterium being administered locally in the gut of the subject.
  • Hepcidin is an hyposideremic hormone made primarily by the liver.
  • recent reports revealed extra-hepatic functional sources of hepcidin, acting in a paracrine/autocrine manner, including the intestine the site of dietary iron absorption.
  • the inventors generated transgenic mice overexpressing the peptide specifically in this tissue (Fig 1C). These mice exhibit, at one month of age, a severe hyposideremia (Fig 2C-D), along with decreased haematological indices (Fig 2H-K) and hair loss (Fig 2A).
  • liver hepcidin gene expression was drastically reduced (Fig 2F) but hepcidin levels were significantly increased in the plasma of Hampl KI ' Int mice compared to controls (Fig 3H).
  • intestinal hepcidin made by the transgenic mice had no effect on intestinal FPN (Fig 3A), but, in contrast, induced a striking down-regulation of DMT1 protein at the apical side of the enterocyte (with no change in mRNA levels) (Fig 3B-C).
  • Intestinal hepcidin can be produced in the apical side (Fig 3G) suggesting the direct role of apical hepcidin on DMT1.
  • hepcidin 25 was injected in isolated duodenal loops. Two hours post injection, DMT1, but not FPN, was found decreased in comparison to PBS injection (Fig 3J).
  • probiotics engineered recombinant lactic acid bacteria
  • Fig 4A probiotics
  • the present invention relates to a method for decreasing iron overload in a subject in need thereof, comprising administering locally in the gut of the subject a therapeutically effective amount of hepcidin.
  • the term "subject” or “patients” refers to a human or another mammal (e.g., primate, dog, cat, goat, horse, pig, mouse, rat, rabbit, and the like), that has or is susceptible to have iron overload associated disease, and in particular P-thalassemia or haemochromatosis.
  • the subject is a human being.
  • the term "subject” does not denote a particular age, and thus encompasses children, teenagers, and adults.
  • the present invention relates to a method for preventing or treating an iron overload associated disease in a subject in need thereof, comprising administering locally in the gut of the subject a therapeutically effective amount of hepcidin.
  • the present invention relates to hepcidin for use for preventing or treating an iron overload associated disease in a subject in need thereof, wherein said hepcidin is administered locally in the gut of the subject.
  • the hepcidin inhibits Divalent Metal Transporter 1 (DMT1).
  • DMT1 Divalent Metal Transporter 1
  • the hepcidin is an inhibitor of the Divalent Metal Transporter 1 (DMT1).
  • DMT1 Divalent Metal Transporter 1
  • DMT1 Divalent Metal Transporter 1
  • DMT1 has its general meaning in the art and refers to the major iron transporter and contributes non-heme iron uptake in most types of cell. DMT1 is present at the apical side of the enterocyte. Intestinal DMT1 is critical for iron absorption.
  • hepcidin inhibits the activity of DMT1, i.e iron absorption.
  • hepcidin regulates DMT1 at the post-transcriptional level.
  • iron overload associated disease refers to a group of diseases and/or disorders which are generally associated with abnormally low levels of serum hepcidin and includes diseases wherein aberrant iron metabolism directly causes the disease, or wherein iron blood levels are dysregulated causing disease, or wherein iron dysregulation is a consequence of another disease, or wherein diseases can be treated by modulating iron levels, and the like. More specifically, a disease of iron metabolism according to this disclosure includes iron overload diseases, disorders of iron biodistribution, other disorders of iron metabolism and other disorders potentially related to iron metabolism, etc.
  • Diseases of iron metabolism include hemochromatosis, HFE mutation hemochromatosis, ferroportin mutation hemochromatosis, transferrin receptor 2 mutation hemochromatosis, hemojuvelin mutation hemochromatosis, hepcidin mutation hemochromatosis, juvenile hemochromatosis, neonatal, hemochromatosis, hepcidin deficiency, thalassemia, thalassemia intermedia, alpha thalassemia, beta thalassemia, African iron overload, hyperferritinemia, ceruloplasmin deficiency, atransferrinemia, congenital dyserythropoietic anemia.
  • the iron overload associated disease is selected from the list consisting of hemochromatosis (adult and juvenile forms of hereditary hemochromatosis), iron- loading anemia and chronic liver diseases including alcoholic liver diseases, non-alcoholic steatohepatitis (NASH) and chronic hepatitis B and C.
  • hemochromatosis adult and juvenile forms of hereditary hemochromatosis
  • iron- loading anemia and chronic liver diseases including alcoholic liver diseases, non-alcoholic steatohepatitis (NASH) and chronic hepatitis B and C.
  • NASH non-alcoholic steatohepatitis
  • Iron loading anaemia also called “secondary iron overload” refers to diseases characterized by anaemia, high serum iron, transferrin saturation and ferritin values, and haemosiderin deposits in parenchymal cells and reticuloendothelial tissue with or without organ dysfunction.
  • the iron-loading anemia is selected from the list consisting of alpha-thalassemia, P-thalassemia, congenital dyserythropoietic anemias, myelodysplastic syndrome (MDS).
  • the iron overload associated disease is hemochromatosis or P-thalassemia.
  • hemochromatosis refers to a genetic disorder characterized by excessive intestinal absorption of dietary iron, resulting in a pathological increase in total body iron stores.
  • type 1, 2 (2A, 2B), 3, 4 and 5 all caused by mutated genes.
  • Hereditary hemochromatosis type 1 is the most frequent, and unique related to the HFE gene. Early diagnosis and treatment is critical to prevent complications from the disorder. Indeed, Too much iron is toxic to the body and over time the high levels of iron can damage tissues and organs and lead to cirrhosis, liver cancer, heart problems, arthritis or diabetes.
  • Beta thalassemias refers to a group of inherited blood disorders. They are forms of thalassemia caused by reduced or absent synthesis of the beta chains of hemoglobin that result in variable outcomes ranging from severe anemia to clinically asymptomatic individuals. Global annual incidence is estimated at one in 100,000 (Galanello, et al (2010). Orphanet J Rare Dis. 5: l l 21 ). Beta thalassemias occur due to malfunctions in the hemoglobin subunit beta or HBB. The severity of the disease depends on the nature of the mutation.
  • HBB blockage over time leads to decreased beta-chain synthesis and accumulation of alpha chains causing premature apoptosis of erythroid precursors.
  • the body's inability to construct new beta-chains leads to the underproduction of HbA (adult hemoglobin).
  • HbA adult hemoglobin
  • Reductions in HbA available overall to fill the red blood cells leads to microcytic anemia.
  • Microcytic anemia ultimately develops in respect to inadequate HBB protein for sufficient red blood cell functioning. Due to this factor, the subject may require blood transfusions to make up for the blockage in the beta-chains. Repeated blood transfusions cause severe problems associated with iron overload(Galanello, et al (2010). Orphanet J Rare Dis. 5: l l 21 ).
  • P -thalassemia is not medically curable except for the use of blood transfusion, although frequent blood transfusions that lead to or potentiate iron overload. Accordingly there is a medical need to specifically treat B-thalassemia subject with new therapeutic approach.
  • treating refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of subject at risk of contracting the disease or suspected to have contracted the disease as well as subject who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
  • the treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
  • the treatment of the disorder may consist in reducing iron overload caused by low hepcidin expression in conditions such as P-thalassemia, haemochromatosis, congenital dyserythropoietic anaemia and myelodysplastic syndromes (MDS). Most preferably, such treatment leads to the complete depletion of the pathological features as observed in iron overload associated disease.
  • therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
  • a therapeutic regimen may include an induction regimen and a maintenance regimen.
  • the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
  • An induction regimen may employ (in part or in whole) a “loading regimen”, which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
  • a “loading regimen” or “maintenance period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a subject during treatment of an illness, e.g., to keep the subject in remission for long periods of time (months or years).
  • a maintenance regimen may employ continuous therapy (e.g., administering a drug at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., pain, disease manifestation, etc.]).
  • continuous therapy e.g., administering a drug at regular intervals, e.g., weekly, monthly, yearly, etc.
  • intermittent therapy e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., pain, disease manifestation, etc.]).
  • a “therapeutically effective amount” is intended for a minimal amount of active agent which is necessary to impart therapeutic benefit to a patient.
  • a “therapeutically effective amount of the active agent” to a patient is an amount of the active agent that induces, ameliorates or causes an improvement in the pathological symptoms, disease progression, or physical conditions associated with the disease affecting the patient. It will be understood that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts. For example, it is well known within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated.
  • a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient.
  • An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 100 mg/kg of body weight per day.
  • Hepcidin has its general meaning in the art and was first identified as a liver-derived antimicrobial peptide.
  • the human hepcidin gene encodes an 84- residue prepropeptide that contains a 24-residue N-terminal signal peptide that is subsequently cleaved to produce pro-hepcidin.
  • Pro-hepcidin is then processed to produce a mature 25-amino acid hepcidin (Jordan et al., The Journal of Biological chemistry, 2009,284, 36, 24155-24167).
  • Hepatic hepcidin is the regulator of iron homeostasis (as disclosed by Lesbordes-Brion et al., Blood, 2006, 108, 1402-1405 and Nicolas et al., Proc. Natl. Acad. Sci. U.S. A., 2002, 99, 4596-4601).
  • Hepcidin is a hypoferremic hormone; it binds to and degrades ferroportin (FPN, the only iron exporter known to date), thereby decreasing the iron levels in the circulation. Since its expression can be induced by inflammation (Nicolas et al., J. Clin. Invest., 2002, 110, 1037-1044), hepcidin has been proposed as an important mediator of Anemia of Chronic disease (ACD), also known as Anemia of Inflammation (Al).
  • ACD Anemia of Chronic disease
  • Al Anemia of Inflammation
  • Hepcidin when used in the context of the present invention, should be understood broadly, it encompasses the mature forms of hepcidin, variants and fragments thereof having same biological activity (regulation of DMT 1 at the apical side of the enterocyte in order to limit iron overload) as well as a nucleic acid encoding said mature forms of hepcidin, variants or fragments.
  • Biological activity of mature form of hepcidin can be measured for example by iron and iron transporters DMT1 quantification (see experimental section and figure 3).
  • Hepcidin can be in the form of the known human 20, 22 or 25-amino acid peptide or of a functional fragment or variant thereof.
  • hepcidin structure is permissive for mutations (Nemeth et al., Blood, 2006, 107, 30 328-333).
  • the fragments of hepcidin according to the present invention comprise 10, 15, 20, 22 or 25 amino acids.
  • Precursors of said mature forms of hepcidin, i.e. prohepcidin and preprohepcidin and nucleic acids encoding said precursors can also be used.
  • Hepcidin encompasses also the analogues/variant of hepcidin which mimic the hepcidin activity of Hepcidin -25, the bioactive human 25-amino acid form.
  • These analogues can be known in the art as “mini-hepcidins” and include PR73 that mimics the first 9 amino residue of hepcidin and analogues described in WO2015200916.
  • Hepcidin or nucleic acid encoding thereof suitable for use according to the invention are vertebrate, preferably mammalian, homologous of mature forms of human hepcidin or precursors thereof.
  • Known vertebrate homologous of human hepcidin include for instance rat hepcidin, mouse hepcidin, trout hepcidin.
  • the hepcidin is a mature form of human hepcidin (UniProtKB - P81172).
  • the protein sequence of said human hepcidin may be found in NCBI database with the following access numbers: mRNA NM_ NM_021175, and protein_id: NP_ NP 066998 (hepcidin preproprotein).
  • Chimeric polypeptides, comprising the sequence of a mature form of hepcidin, can also be used.
  • the hepcidin comprises or consists of the sequence having at least 50%, 60%, 70%, 80% or more particularly 90% sequence identity with the amino acid sequence selected in the group consisting of SEQ ID NO: 1, SEQ ID NO:2 or SEQ ID NO:3.
  • a first amino acid sequence having at least 90% of identity with a second amino acid sequence means that the first sequence has 90; 91; 92; 93; 94; 95; 96; 97; 98; 99 or 100% of identity with the second amino acid sequence.
  • Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar are the two sequences.
  • Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith and Waterman, Adv. Appl. Math., 2:482, 1981; Needleman and Wunsch, J. Mol. Biol., 48:443, 1970; Pearson and Lipman, Proc. Natl. Acad. Sci.
  • the alignment tools ALIGN Myers and Miller, CAB IOS 4: 11-17, 1989
  • LFASTA Pearson and the University of Virginia, fasta20u63 version 2.0u63, release date December 1996.
  • the hepcidin comprises a sequence as set forth in SEQ ID NO: 3 (Hepcidin - 25)
  • the invention also encompasses the use of functional equivalents of the above-defined hepcidin.
  • Functional equivalents are herein defined as peptide variants, having the same functional biological activity as the mature forms of hepcidin.
  • polypeptides and nucleic acids can be obtained by classical methods known in themselves.
  • the 20 amino-acids and 25 amino-acids forms of hepcidin can be obtained from plasma or from urine, as disclosed by KRAUSE et al. ( FEBS Lett., 2000, vol. 480, 147-150) ; or PARK et al. (J. Biol. Chem., 2001, vol. 276,7806-7810).
  • they can be obtained by culturing cells expressing hepcidin, and recovering said polypeptide from the cell culture.
  • said cells are host cells transformed by a nucleic acid encoding one of the polypeptides defined above.
  • hepcidin can be obtained by classical sequential procedures for hepcidin purification from total human plasma.
  • ADDO L et al Int J Hematol. 2016 Jan;103(l):34-43 purified 3 isoform of human hepcidin by using liquid chromatography-tandem mass spectrometry (LC-tandem MS).
  • LC-tandem MS liquid chromatography-tandem mass spectrometry
  • LC-MS/MS liquid chromatography coupled with tandem mass spectrometry
  • hepcidin can be a recombinant hepcidin as described in Gagliardo et al., FEBS J. 2008 Aug;275(15):3793-803.
  • a nucleic acid encoding hepcidin can for instance be obtained from a genomic or cDNA library of a vertebrate, using suitable primers able to hybridize selectively with said nucleic acids. It can also be obtained by the classical techniques of polynucleotide synthesis.
  • hepcidin has at least 50%, preferably, at least 60%, preferably, at least 70%, preferably, at least 80%, preferably, at least 85% more preferably at least 90%, more preferably at least 95% and even more preferably at least 99% identity with human mature form of hepcidin.
  • administering refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., hepcidin) into the subject, such as by oral, mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery and/or any other method of physical delivery described herein or known in the art.
  • a disease, or a symptom thereof is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof.
  • administration of the substance typically occurs before the onset of the disease or symptoms thereof.
  • the hepcidin in order to be administered directly to the intestine (gut), the hepcidin is administered to the subject orally (including buccal and sublingual administration), rectally or topically (intracolic administration).
  • the hepcidin according to the invention can be in the form of a sustained release composition, or can be produced by a recombinant bacteria so as to deliver the hepcidin directly to the intestine.
  • the hepcidin is administered in intestinal lumen of the subject.
  • the hepcidin is administered in the gut of the subject by using a gut lumen-targeted delivery system.
  • gut lumen-targeted delivery system refers to delivery system has been developed for specific delivery and controlled release of cargos to gut area, i.e the gut lumen.
  • Gut lumen-targeted delivery system not only protects the cargos from premature degradation and controls the release of the cargos in the gut area, greatly improving its bioavailability and reducing side effects, but also has better patient compliance and less interference by a variety of immune cells and cytokine in circulatory system.
  • direct oral administration of bioactive agents (active substances) in the intestine (gut) can be fraught with problems due to the complex and harsh gastrointestinal environment.
  • the strong acidic environment in the stomach inactivates many bioactive substances, while digestive enzymes in the small intestine further decompose and absorb them
  • Gut lumen-targeted delivery system can include Gut lumen-targeted oral delivery system as described in Liu, J. et al. 20 and/or and/or probiotic bacterium (or food-grade bacterium) as described in Benbouziane B. et al. 2013. 17 and/or mucosal delivery system including lipid carrier delivery systems such as micelles, liposomes and nanoparticle-based oral delivery systems as described in Zhang, M. et al 22 .
  • lipid carrier delivery systems such as micelles, liposomes and nanoparticle-based oral delivery systems as described in Zhang, M. et al 22 .
  • the hepcidin is administered in the gut of the subject by using a food-grade bacterium or probiotic bacterium.
  • object of the invention relates to a recombinant food-grade bacterium comprising a gene coding for hepcidin for use for preventing or treating iron overload associated disease in a subject in need thereof, wherein said recombinant food-grade bacterium is being administered locally in the gut of the subject.
  • another aspect of the invention relates to a method for preventing or treating iron overload associated disease in a subject in need thereof, comprising administering to said subject a food-grade bacterium comprising a gene coding for hepcidin, said food-grade bacterium is used to deliver directly the hepcidin in the gut of said subject.
  • another aspect of the invention relates to hepcidin for use preventing or treating iron overload associated disease in a subject in need thereof, wherein a food-grade bacterium comprising a gene coding for said hepcidin is used to deliver directly the hepcidin in the gut.
  • the recombinant food-grade bacterium comprising a gene coding for hepcidin having sequence set forth as SEQ ID NO:3.
  • the term “food-grade bacterium” denotes a bacterium that is widely used in fermented foods and possesses a perfect safety profile recognized by the GRAS (Generally Recognized As Safe) and QPS (Qualified Presumption of Safety) status in USA and European Community, respectively. Such bacterium can be safely in functional foods or food additives with allegations concerning maintain in good health and well-being or prevention of disease.
  • the term food-grade bacterium includes also probiotic bacterium.
  • probiotic bacterium denotes a bacterium which ingested live in adequate quantities can exert beneficial effects on the human health. They are now widely used as a food additive for their health-promoting effects. Most of the probiotic bacteria are Lactic Acid Bacterium (LAB) and among them strains of the genera Lactobacillus spp. and Bifidobacterium spp. are the most widely used probiotic bacteria.
  • LAB Lactic Acid Bacterium
  • the food-grade bacterium strain according to the invention is a Lactococcus lactis strain or a Lactobacillus casei strain or a Lactococcus lactis htrA strain [Poquet et al., 2000] or a Lactobacillus plantarum strain or a Bifidobacterium longum strain.
  • the food-grade bacterium strain according to the invention is a Lactobacillus lactis strain.
  • the food-grade bacterium strain according to the invention is a lactic acid bacteria.
  • lacid acid bacteria are well known as mucosal delivery vehicles, as described in Benbouziane B. et al. 2013. 17
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain.
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain comprising a controlled expression system.
  • controlled expression system refers to a system comprising inducible promoter enabling to regulates the expression of the gene of interest and to ensure that the protein of interest are coded when and where they are needed.
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain comprising a Stress-Inducible Controlled Expression (SICE) system.
  • SICE Stress-Inducible Controlled Expression
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain comprises the groESL operon promoter.
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain comprising a Stress-Inducible Controlled Expression (SICE) system as described in Benbouziane B. et al. 2013. 17
  • the food-grade bacterium strain according to the invention is an engineered Lactobacillus lactic strain comprising the groESL operon promoter and a gene coding for hepcidin having sequence set forth as SEQ ID NO:3.
  • the recombinant food-grade bacterium according to the invention is administered to the gut’s subject preferably orally (including buccal and sublingual administration), rectally or topically (intracolic administration).
  • compositions of the invention are provided.
  • hepcidin and recombinant food-grade bacterium as described above may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions.
  • the present invention relates to a pharmaceutical composition for use in the prevention or treatment of iron overload associated disease in a subject in need thereof, said pharmaceutical composition comprises hepcidin or a recombinant food-grade bacterium as defined above.
  • the present invention also relates to a pharmaceutical composition for use in the prevention or treatment of iron overload associated disease in a subject in need thereof, said pharmaceutical composition comprises i) hepcidin or a recombinant food-grade bacterium as defined above, and ii) a pharmaceutically acceptable carrier.
  • “Pharmaceutically” or “pharmaceutically acceptable” refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate.
  • a pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
  • compositions are administered to a patient already suffering from a disease, as described, in an amount sufficient to cure or at least partially stop the symptoms of the disease and its complications.
  • An appropriate dosage of the pharmaceutical composition is readily determined according to any one of several well-established protocols. For example, animal studies (for example on mice or rats) are commonly used to determine the maximal tolerable dose of the bioactive agent per kilogram of weight. In general, at least one of the animal species tested is mammalian. The results from the animal studies can be extrapolated to determine doses for use in other species, such as humans for example. What constitutes an effective dose also depends on the nature and severity of the disease or condition, and on the general state of the patient's health.
  • the antagonist contained in the pharmaceutical composition can be administered in several dosages or as a single dose until a desired response has been achieved.
  • the treatment is typically monitored and repeated dosages can be administered as necessary.
  • the active principle in the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings.
  • Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.
  • the appropriate unit forms of administration include forms for oral administration, such as tablets, gelatine capsules, powders, granules and solutions or suspensions to be taken orally, forms for sublingual and buccal administration, aerosols, implants, forms for subcutaneous, intramuscular, intravenous, intranasal or intraocular administration and forms for rectal administration.
  • the active principle is generally formulated as dosage units containing from 0.5 to 1000 mg, preferably from 1 to 500 mg, more preferably from 2 to 200 mg of said active principle per dosage unit for daily administrations.
  • a wetting agent such as sodium laurylsulfate can be added to the active principle optionally micronized, which is then mixed with a pharmaceutical vehicle such as silica, gelatine, starch, lactose, magnesium stearate, talc, gum arabic or the like.
  • a pharmaceutical vehicle such as silica, gelatine, starch, lactose, magnesium stearate, talc, gum arabic or the like.
  • the tablets can be coated with sucrose, with various polymers or other appropriate substances or else they can be treated so as to have a prolonged or delayed activity and so as to release a predetermined amount of active principle continuously.
  • a preparation in the form of gelatin capsules is obtained by mixing the active principle with a diluent such as a glycol or a glycerol ester and pouring the mixture obtained into soft or hard gelatine capsules.
  • a preparation in the form of a syrup or elixir can contain the active principle together with a sweetener, which is preferably calorie-free, methyl-paraben and propylparaben as an antiseptic, a flavoring and an appropriate color.
  • the water-dispersible powders or granules can contain the active principle mixed with dispersants or wetting agents, or suspending agents such as polyvinyl-pyrrolidone, and also with sweeteners or taste correctors.
  • the active principle can also be formulated as microcapsules or microspheres, optionally with one or more carriers or additives.
  • implants can be used. These can be prepared in the form of an oily suspension or in the form of a suspension of microspheres in an isotonic medium.
  • Hepcidin or food-grade bacterium according to the invention can be administered by any suitable route of administration.
  • hepcidin or food-grade bacterium according to the invention can be administered by oral (including buccal and sublingual), rectal, nasal, topical (intracolic), pulmonary, vaginal, or parenteral (including intramuscular, intra-arterial, intrathecal, subcutaneous and intravenous) administration.
  • the hepcidin or food-grade bacterium of the present invention may be formulated in a wide variety of oral administration dosage forms.
  • preparation is intended to include the formulation of the active compound with an encapsulating material as carrier, providing a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is in association with it.
  • encapsulating material as carrier
  • cachets and lozenges are included. Tablets, powders, capsules, pulls, cachets, and lozenges may be as solid forms suitable for oral administration.
  • Other forms suitable for oral administration include liquid form preparations including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, or solid form preparations which are intended to be converted shortly before use to liquid form preparations.
  • Emulsions may be prepared in solutions, for example, in aqueous propylene glycol solutions or may contain emulsifying agents, for example, such as lecithin, sorbitan monooleate, or acacia.
  • Aqueous solutions can be prepared by dissolving the active component in water and adding suitable colorants, flavours, stabilizers, and thickening agents.
  • Aqueous suspensions can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.
  • Solid form preparations include solutions, suspensions, and emulsions, and may contain, in addition to the active component, colorants, flavours, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.
  • the hepcidin or food-grade bacterium of the present invention of the present invention may be formulated for administration as suppositories.
  • a low melting wax such as a mixture of fatty-acid glycerides or cocoa butter is first melted and the active component is dispersed homogeneously.
  • FIGURES are a diagrammatic representation of FIGURES.
  • FIG. 1 Hepcidin is expressed all along the intestine.
  • A. Hampl expression mRNA (qPCR) in the gut of 8-10 weeks old C57B16j mice (n 4).
  • B. Hampl expression mRNA (qPCR) along the duodenum villi in 8-10 weeks old C57B16j mice (n 4).
  • Figure 2_ Hampl Kl-Vil mice developped en iron deficiency anemia
  • Blood count parameters Red blood cells (10 12 /L), Hemoglobine (g/dL), Hematocrit (%), Mean Corpusula Volume (MCV fL) and Mean Corpuscular Hemoglobin (MCH pg). All of analyses was done on male and female Hampl CMV ' 1O X -STOP-1OX ( rounc [) anc [ Hamp 1 KI ' V11 (square) aged of 26 days old. Mean +/- SD. Unpaired Mann- Whitney test used : *P ⁇ 0.05, **P ⁇ 0.01 and ***P ⁇ 0.001.
  • Figure 3 Apical hepcidin decreased iron absorption via downregulation of DMT1
  • Lac-pGroESL and Lac-hepc constructs and quantification of hepcidin production (n 3) by ELISA after stimulation of bacteria at 37°C for 2 hours.
  • H-K Tissus iron content (pg/g) in liver, pancreas, spleen and heart respectively.
  • Hampl KLvil mice To generate Hampl KLvil mice, we first generated a Rosa26-pCAG-LSL-Hamp mouse line by homologous recombination Knock-in technology into Rosa26 locus.
  • Hampl cDNA was inserted downstream the cytomegalovirus (CMV), immediate enhancer/p-actin (CAG) promoter and a loxP-STOP-loxP (LSL) cassette.
  • CMV cytomegalovirus
  • CAG immediate enhancer/p-actin
  • LSL loxP-STOP-loxP
  • the vector was transfected into embryonic stem cell (ESC) line and the selection was made in G418.
  • ES cell clones were screened for proper homologous recombination by DNA-PCR before injection into BALB/c blastocysts. Chimeric male mice were crossed with C57BL/6N female mice to obtain the ROSAHampl Fl mice.
  • the identification of Fl mice was performed by PCR of tail DNA using primers that amplify a 310 bp band in case of the mutant allele, and primers that amplify a 192 bp band in the case of the WT Rosa Allele.
  • ROSAHampl Fl mice don’t express the Hampl cDNA but after breeding with B6.Cg-Tg(Vill-cre)1000Gum/J mice (Jackson Laboratory) 15 , the LSL cassette were deleted and Hampl mRNA was expressed in the gut epithelial cells.
  • Hemochromatosis mouse model (Hampl Allver ) was generated by our laboratory as described before 16
  • the stable iron isotope 57 Fe ( 57 Fe at 96% enrichment, MFE57M, Eurisotop) was used as tracer.
  • the procedure was adapted from Fiorito et al. 17 days old Ham P 1CMV - lox - STOp - lox anc [ Hamp l Klv11 mice were force-fed with a solution of 20mM 57 FeSO4, 5.38mM ascorbic acid and 10%sucrose. 90min after oral administration, mice were anesthetized, perfused and their duodenum collected. Iron isotope measurement was performed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) on the Nexion 5000® spectrometer (Perkin Elmer, Les Ulis, France).
  • ICP-MS Inductively Coupled Plasma Mass Spectrometry
  • mice 7 weeks old male C57B16/J mice were anesthetized with pentobarbital. Next, mice were open and the duodenum was ligatured twice, first at the pylore and 3cm after pylore. Before to finish the second ligature, 400pL PBS or 400pL hepcidin-25 (IpM) were injected in the duodenum lumen. Mice have been closed during 2 hours. Mice were maintained under isoflurane during the surgery. 2 hours later, sleeping mice were open and the duodenum were collected.
  • IpM hepcidin-25
  • Lac-pGroESL and Lac-hepc was grown in agar-M17 medium (Difco Laboratories) supplemented with 0.5% glucose and antibiotic Chloramphenicol (Cm, lOpg/mL). The next day, one colony was resuspended in 5mL of M17 + 0.5% glucose + Cm and growing at 30°C all day. Ten hours later, 5mL of preculture was added to IL of M17 + 0.5% glucose + Cm at 30°C overnight. DO (600nm) was measured after overnight incubation to determine L. lactis concentration. Bacteria were centrifugated at 6,000 rpm, 4°C, 15min.
  • Hampl fl/fl and Hampl Allver were weaned at 28 days old and directly treated with frozen aliquot of Lac-pGroESL or Lac-hepc. Male and female are included. Mice were treated every day (at 6 pm) by oral gavage with 5xlO 9 CFU of Lac-pGroESL or Lac-hepc. Mice were separated in cage by group sex and treatment. They received food and water ad libitum.
  • RNA extraction, reverse transcription and quantitative PCR have been performed as previously described 16 .
  • Mouse samples were normalized to the threshold cycle value for 36B
  • Epithelial cells from duodenum were harvested and resuspended in a solution of 0.25 M sucrose/0.03 M histidine (pH 7.2) supplemented with l x complete inhibitor cocktail EDTA- free (Roche) (Volume is 4 times the weight of tissue). After a 30min incubation on ice, cells were subjected to brief sonication and centrifugation at 6,000 g for 10 min. To obtain crude membrane fraction the supernatant was subsequently isolated by ultracentrifugation at 41 000 rpm (TLA-45 rotor; Beckman-Coulter) for 1 hour. Protein concentration was determined using the PierceTM BCA Protein Assay Kits (23225, Thermo Scientific).
  • Lysates containing 15pg of proteins were subjected to standard methos of Western blot.
  • DMT1 primary antibody gift of Francois Cannone-Hergaux, CNRS [ICSN], Gif-sur-Yvette, France
  • FPN primary antibody NBP1-21502, Novus
  • Loading control P-actin (A5316, Sigma Aldrich at 1 :5000) antibodies were also used. Immunoblots were quantified using ImageJ software.
  • Plasma and iron tissues (heart, liver, pancreas and spleen) levels were quantified colorimetrically by a previously described method 18 . Hematological parameters were determined using a OX-560 RET automatic analyzer (Baliodiasgnotics). Perl’s staining
  • Liver tissues were fixed in 4% formaldehyde and embedded in paraffin. Sections were cut at a thickness of 4 pm. Following dewaxing in xylene baths and rehydratation, sections were either stained with Peris’s Prussian blue and nuclear fast red counterstain.
  • mice The serum from mice were directly used in Hepcidin Murine-CompeteTM ELISA (HMC-001, Intrinsic Lifesciences) and in mouse TNFa Quantikine ELISA (MTAOOB, R&Dsy stems). Luminal duodenum content was collected in ImL PBS IX supplemented with inhibitor protease (A32965, Thermo Scientific). Tissues was homogenized by vortex and directly used in Hepcidin Murine-CompeteTM ELISA (HMC-001, Intrinsic Lifesciences). After induction of hepcidin production by Lac-hepc and Lac-pGroESL, supernatant was used in mouse hepcidin ELISA (S-1465, BMA Biomedicals).
  • Hepcidin is expressed by the gut
  • hepcidin- ferroportin axis may be crucial, in pathophysiological conditions, for maintaining iron homeostasis in these organs.
  • hepcidin- ferroportin axis may be crucial, in pathophysiological conditions, for maintaining iron homeostasis in these organs.
  • mice To probe the function of gut-derived hepcidin, we generated a mouse model overexpressing hepcidin in intestinal epithelium. For that, the mouse Hampl gene sequence was inserted into the ROSA26 locus downstream of a strong CMV promoter and a lox-STOP- lox (LSL) sequence (referenced as Hampl CMV ' lox ' STOP ' lox ). After breeding with a transgenic strain expressing the Cre recombinase under the control of the murine villin promoter (Villinl - CRE), the LSL cassette is deleted and Hampl is overexpressed. This newly generated model is referenced as Hamp 1 KI ' V11 (See Methods, Fig 1C).
  • LSL lox-STOP- lox
  • Duodenal lumen hepcidin decreased iron absorption via DMT1 inhibition
  • transgenic hepcidin on DMT1 at the apical side of the enterocyte prompted us to ask whether the transgenic hepcidin peptide could be secreted in the intestinal lumen.
  • Fig 3G we did observe the presence of hepcidin in the intestinal lumen, with a twofold increase of hepcidin in the Hampl KI ' V11 mice as compared to Hampl CMV ' lox ' STOP ' lox mice.
  • duodenal hepcidin may regulate iron absorption by controlling the levels of apical DMT1 and may induce, when overexpressed as in the Hampl KI ' V11 mice, iron deficiency anemia Probiotic L. lactis producing hepcidin decrease iron overload in hemochromatosis mouse model
  • hepcidin can be secreted into the lumen to diminish DMT1 and iron absorption.
  • hepcidin could be a new therapeutic target to inhibit iron absorption in case of pathologies linked to iron hyperabsorption.
  • Hereditary hemochromatosis is a genetic disorder characterized by an iron overload. In these patients, hepatic hepcidin is abnormally low. This defect in liver hepcidin expression induces an iron hyperabsorption.
  • probiotics enable to produce hepcidin as therapeutic tools.
  • the most common probiotic strain belongs to Lactococcus, Lactobacillus and Bifidobacterium.
  • Lactococcus lactis (L. lactis), considered a "neutral strain”, is a microbial model widely used as an expression system for molecules of interest 19 . It has the advantage of being a GRAS (Generally Recognized As Safe) bacterium.
  • L. lactis recombinant for hepcidin (Lac-hepc).
  • the murine hepcidin gene was cloned into the Histidine-tagged pSICE plasmid under the control of the L. lactis GroESL promoter (Fig 4A). This promoter is inducible by stress, notably heat and pH, and will thus enable the induction of hepcidin in vivo.
  • the plasmid was sequenced and transformed into the L. lactis strain. We verified hepcidin secretion in vitro in the culture supernatant after heat induction (Fig 4A).
  • mice After validation of production of hepcidin by Lac-hepc, we administered by oral gavage of L-hepc to hemochromatosis mice (Hampl Allv ) and we showed a decrease in membrane duodenal DMT 1 expression as early as 3 hours (Fig 4B).
  • Brasse-Lagnel, C. et al Intestinal DMT1 cotransporter is down-regulated by hepcidin via proteasome internalization and degradation. Gastroenterology 140, 1261-1271. el (2011).

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Abstract

L'hepcidine est une hormone hyposidérémique principalement produite par le foie. Pour répondre au rôle de l'hepcidine produite spécifiquement par l'intestin dans l'abaissement du fer sérique, les inventeurs ont généré des souris transgéniques surexprimant le peptide spécifiquement dans ce tissu. Ces souris présentent, à un mois d'âge, une hyposidérémie sévère, ainsi que des indices hématologiques réduits et une perte de cheveux. Mécaniquement, ils ont montré que l'hepcidine intestinale produite par les souris transgéniques n'avait aucun effet sur la ferroportine intestinale, mais, au contraire, a induit une régulation à la baisse remarquable de la protéine du transporteur de métal divalent 1 (DMT1) au niveau du côté apical de l'entérocyte. L'hepcidine intestinale peut être produite dans le côté apical suggérant le rôle direct d'hepcidine apicale sur DMT1. Pour confirmer la capacité thérapeutique de l'hepcidine sur la régulation du DMT1, les inventeurs ont développé des probiotiques (bactéries lactiques recombinantes modifiées), capables de délivrer de l'hepcidine directement dans la lumière de l'intestin. Ils administrent par voie orale quotidiennement ces probiotiques dans un modèle de souris atteintes d'hémochromatose, après 28 jours de traitements ils observent une diminution de surcharge en fer. Ainsi, la présente invention concerne une méthode de prévention ou de traitement d'une maladie associée à une surcharge en fer chez un sujet en ayant besoin, comprenant l'administration locale d'hepcidine dans l'intestin du sujet.
PCT/EP2025/060984 2024-04-24 2025-04-23 Méthodes de traitement de maladies associées à une surcharge en fer par administration locale d'hepcidine dans l'intestin Pending WO2025224128A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011086172A1 (fr) * 2010-01-14 2011-07-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Bactéries probiotiques recombinantes pour la prévention et le traitement du syndrome abdominal inflammatoire (ibd) et du syndrome du côlon irritable (ibs)
WO2015200916A2 (fr) 2014-06-27 2015-12-30 Protagonist Therapeutics, Inc. Analogues d'hepcidine et de mini-hepcidine, et leurs utilisations
WO2016109363A1 (fr) * 2014-12-29 2016-07-07 The Regents Of The University Of California Peptides hepcidine s-alkylés et leurs procédés de préparation et d'utilisation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011086172A1 (fr) * 2010-01-14 2011-07-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Bactéries probiotiques recombinantes pour la prévention et le traitement du syndrome abdominal inflammatoire (ibd) et du syndrome du côlon irritable (ibs)
WO2015200916A2 (fr) 2014-06-27 2015-12-30 Protagonist Therapeutics, Inc. Analogues d'hepcidine et de mini-hepcidine, et leurs utilisations
WO2016109363A1 (fr) * 2014-12-29 2016-07-07 The Regents Of The University Of California Peptides hepcidine s-alkylés et leurs procédés de préparation et d'utilisation

Non-Patent Citations (46)

* Cited by examiner, † Cited by third party
Title
"Council of Europe", EUROPEAN CONVENTION FOR THE PROTECTION OF LABORATORY ANIMALS, 1991
ABBAS ET AL., ANAL BIOANAL CHEM, 18 April 2018 (2018-04-18)
ADDO L ET AL., INT J HEMATOL, vol. 103, no. 1, January 2016 (2016-01-01), pages 34 - 43
ALTSCHUL ET AL., NAT. GENET., vol. 6, 1994, pages 119 - 129
BENBOUZIANE, B. ET AL.: "Development of a Stress-Inducible Controlled Expression (SICE) system in Lactococcus lactis for the production and delivery of therapeutic molecules at mucosal surfaces", J BIOTECHNOL, vol. 168, 2013, pages 120 - 129
BERM�DEZ-HUMAR�N LUIS G ET AL: "Live bacterial biotherapeutics in the clinic", NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 36, no. 9, 1 October 2018 (2018-10-01), pages 816 - 818, XP036929666, ISSN: 1087-0156, [retrieved on 20181001], DOI: 10.1038/NBT.4248 *
BERMUDEZ-HUMARAN, L. G.LANGELLA, P.: "Live bacterial biotherapeutics in the clinic.", NAT BIOTECHNOL, vol. 36, 2018, pages 816 - 818, XP036929666, DOI: 10.1038/nbt.4248
BRASSE-LAGNEL, C. ET AL.: "Intestinal DMT1 cotransporter is down-regulated by hepcidin via proteasome internalization and degradation", GASTROENTEROLOGY, vol. 140, 2011, pages 1261 - 1271
CHASTON, T. ET AL.: "Evidence for differential effects of hepcidin in macrophages and intestinal epithelial cells", GUT, vol. 57, 2008, pages 374 - 382
CORPET ET AL., NUC. ACIDS RES., vol. 16, 1988, pages 10881 - 10890
DESCHEMIN, J.-C.MATHIEU, J. R. R.ZUMERLE, S.PEYSSONNAUX, C.VAULONT, S.: "Pulmonary Iron Homeostasis in Hepcidin Knockout Mice", FRONT PHYSIOL, vol. 8, 2017, pages 804
GAGLIARDO ET AL., FEBS J, vol. 275, no. 15, August 2008 (2008-08-01), pages 3793 - 803
GALANELLO ET AL., ORPHANET J RARE DIS, vol. 5, 2010, pages 1121
GALANELLO RORIGA R.: "Beta-thalassemia", ORPHANET J RARE DIS, vol. 5, 21 May 2010 (2010-05-21), pages 11, XP021081722, DOI: 10.1186/1750-1172-5-11
GANZ, T.: "Systemic iron homeostasis", PHYSIOL REV, vol. 93, 2013, pages 1721 - 1741
HIGGINSSHARP, CABIOS, vol. 4, 1989, pages 151 - 153
HIGGINSSHARP, GENE, vol. 73, 1988, pages 237 - 244
HUANG ET AL., COMP. APPLS BIOSCI., vol. 8, 1992, pages 155 - 165
JORDAN ET AL., THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 284, no. 36, 2009, pages 24155 - 24167
KAWABATA HIROSHI ED - ABE MASAHIRO: "The mechanisms of systemic iron homeostasis and etiology, diagnosis, and treatment of hereditary hemochromatosis", INTERNATIONAL JOURNAL OF HEMATOLOGY, ELSEVIER SCIENCE PUBLISHERS, NL, vol. 107, no. 1, 13 November 2017 (2017-11-13), pages 31 - 43, XP036386179, ISSN: 0925-5710, [retrieved on 20171113], DOI: 10.1007/S12185-017-2365-3 *
KEMNA ET AL., CLIN CHEM., vol. 53, no. 4, April 2007 (2007-04-01), pages 620 - 8
KRAUSE ET AL., FEBS LETT, vol. 480, 2000, pages 147 - 150
LAKHAL-LITTLETON, S. ET AL.: "An essential cell-autonomous role for hepcidin in cardiac iron homeostasis", ELIFE, vol. 5, 2016, pages 19804
LEFEBVRE T ET AL., CLIN CHEM LAB MED, vol. 53, no. 10, 1 September 2015 (2015-09-01), pages 1557 - 67
LIU JUNYU ET AL: "Gut lumen-targeted oral delivery system for bioactive agents to regulate gut microbiome", JOURNAL OF FUTURE FOODS, vol. 2, no. 4, 1 December 2022 (2022-12-01), pages 307 - 325, XP093205606, ISSN: 2772-5669, DOI: 10.1016/j.jfutfo.2022.08.003 *
LIU, J. ET AL.: "Gut lumen-targeted oral delivery system for bioactive agents to regulate gut microbiome", JOURNAL OF FUTURE FOODS, pages 202218
MADISON, B. B. ET AL.: "Cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine", J BIOI CHEM, vol. 277, 2002, pages 33275 - 33283
MALERBA, M. ET AL.: "Epidermal hepcidin is required for neutrophil response to bacterial infection", J CLIN INVEST, vol. 130, 2020, pages 329 - 334
MASTROGIANNAKI, M. ET AL.: "HIF-2q, but not HIF-1α, promotes iron absorption in mice", J CLIN INVEST, vol. 119, 2009, pages 1159 - 1166
MENA, N. P.ESPARZA, A.TAPIA, V.VALDÉS, P.NÚÑEZ, M. T.: "Hepcidin inhibits apical iron uptake in intestinal cells", AM J PHYSIOL GASTROINTEST LIVER PHYSIOL, vol. 294, 2008, pages 192 - 198
NEEDLEMANWUNSCH, J. MOL. BIOL., vol. 48, 1970, pages 443
NEMETH, E. ET AL.: "Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization", SCIENCE, vol. 306, 2004, pages 2090 - 2093
NICOLAS ET AL., J. CLIN. INVEST., vol. 110, 2002, pages 1037 - 1044
NICOLAS ET AL., PROC. NATL. ACAD. SCI., vol. 99, 2002, pages 4596 - 4601
NICOLAS, G. ET AL.: "Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice", PROC NATL ACAD SCI, vol. 98, 2001, pages 8780 - 8785, XP002211395, DOI: 10.1073/pnas.151179498
PARK ET AL., J. BIOL. CHEM., vol. 276, 2001, pages 7806 - 7810
PEARSON ET AL., METH. MOL. BIOL., vol. 24, 1994, pages 307 - 31
PEARSONLIPMAN, PROC. NATL. ACAD. SCI., vol. 85, 1988, pages 2444
PEYSSONNAUX, C. ET AL.: "TLR4-dependent hepcidin expression by myeloid cells in response to bacterial pathogens", BLOOD, vol. 107, no. 30, 2006, pages 3727 - 3732, XP009510781, DOI: 10.1182/blood-2005-06-2259
PIETRANGELO, A.: "Hereditary Hemochromatosis: Pathogenesis, Diagnosis, and Treatment", GASTROENTEROLOGY, vol. 139, 2010, pages 393 - 408
SCHWARTZ, A. J. ET AL.: "Hepcidin sequesters iron to sustain nucleotide metabolism and mitochondrial function in colorectal cancer epithelial cells", NAT METAB, vol. 3, 2021, pages 969 - 982
SMITHWATERMAN, ADV. APPL. MATH., vol. 2, 1981, pages 482
STEINBICKER, A. U.MUCKENTHALER, M. U.: "Out of balance--systemic iron homeostasis in iron-related disorders", NUTRIENTS, vol. 5, 2013, pages 3034 - 3061, XP055949806, DOI: 10.3390/nu5083034
W. R. PEARSON: "Internet Program®", December 1996, THE UNIVERSITY OF VIRGINIA
YAMAJI, S.SHARP, P.RAMESH, B.SRAI, S. K.: "Inhibition of iron transport across human intestinal epithelial cells by hepcidin", BLOOD, vol. 104, 2004, pages 2178 - 2180
ZUMERLE, S. ET AL.: "Targeted disruption of hepcidin in the liver recapitulates the hemochromatotic phenotype", BLOOD, vol. 123, 2014, pages 3646 - 3650

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