WO2018162426A1 - Traitement et diagnostic d'une xérocytose héréditaire - Google Patents

Traitement et diagnostic d'une xérocytose héréditaire Download PDF

Info

Publication number
WO2018162426A1
WO2018162426A1 PCT/EP2018/055373 EP2018055373W WO2018162426A1 WO 2018162426 A1 WO2018162426 A1 WO 2018162426A1 EP 2018055373 W EP2018055373 W EP 2018055373W WO 2018162426 A1 WO2018162426 A1 WO 2018162426A1
Authority
WO
WIPO (PCT)
Prior art keywords
piezol
channel
mutation
inhibitor
mutations
Prior art date
Application number
PCT/EP2018/055373
Other languages
English (en)
Inventor
Hélène GUIZOUARN
Catherine BADENS
Loïc GARCON
Olivier SORIANI
Raphaël RAPETTI-MAUSS
Véronique Picard
Original Assignee
INSERM (Institut National de la Santé et de la Recherche Médicale)
Universite Paris-Sud
Centre Hospitalier Universitaire D'amiens
Universite De Picardie Jules Verne
Université D'aix-Marseille
Assistance Publique Hopitaux De Marseille
Université de Nice Sophia Antipolis
Centre National De La Recherche Scientifique - Cnrs -
Sorbonne Universite
Assistance Publique Hopitaux De Paris
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by INSERM (Institut National de la Santé et de la Recherche Médicale), Universite Paris-Sud, Centre Hospitalier Universitaire D'amiens, Universite De Picardie Jules Verne, Université D'aix-Marseille, Assistance Publique Hopitaux De Marseille, Université de Nice Sophia Antipolis, Centre National De La Recherche Scientifique - Cnrs -, Sorbonne Universite, Assistance Publique Hopitaux De Paris filed Critical INSERM (Institut National de la Santé et de la Recherche Médicale)
Publication of WO2018162426A1 publication Critical patent/WO2018162426A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4174—Arylalkylimidazoles, e.g. oxymetazolin, naphazoline, miconazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/15—Oximes (>C=N—O—); Hydrazines (>N—N<); Hydrazones (>N—N=) ; Imines (C—N=C)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/341—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide not condensed with another ring, e.g. ranitidine, furosemide, bufetolol, muscarine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A61K31/421—1,3-Oxazoles, e.g. pemoline, trimethadione
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4245—Oxadiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4422—1,4-Dihydropyridines, e.g. nifedipine, nicardipine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/661—Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion or mevinphos
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/22—Haematology

Definitions

  • the present invention relates to the treatment and the diagnosis of the hereditary xerocytosis.
  • Water and solute homeostasis is essential for the maintenance of erythrocyte integrity and is controlled via the regulation of monovalent cation content.
  • erythrocytes hydration exist and are characterized by an abnormal permeability of the erythrocyte membrane to sodium and potassium, resulting either in swelling or shrinkage of red cells (Rinehart et al. , 2010).
  • Clinically, these inherited disorders are associated with chronic hemolytic anemia and are due to defects in various transmembrane ion channels or transporters (Da Costa L, et al. , 2013).
  • HX Hereditary xerocytosis
  • DHSt is an autosomal dominant hemolytic anemia characterized by primary erythrocyte (also named red blood cells) dehydration caused by an increase of their permeability to cations (Delaunay J., 2004).
  • HX erythrocytes exhibit decreased total cation and potassium content that are not accompanied by a proportional net gain of sodium and water.
  • HX patients typically exhibit mild to moderate compensated hemolytic anemia, with an increased erythrocyte mean corpuscular hemoglobin concentration and a decreased osmotic fragility, both of which reflect cellular dehydration. Patients may also show perinatal edema and pseudohyperkalemia due to loss of potassium from red cells stored at room temperature.
  • red blood cells A minor proportion of red blood cells (RBCs) appear as stomatocytes on blood films. Complications such as splenomegaly and cholelithiasis, resulting from increased red cell trapping in the spleen and elevated bilirubin levels, respectively, may occur. The course of DHS is also frequently associated with iron overload, which may lead to hepatosiderosis.
  • KCNN4 coding for a calcium activated potassium channel also named Gardos channel in RBCs
  • PIEZOl coding for the non-selective cation channel PIEZOl, activated by mechanical forces.
  • the Gardos channel has been initially described in erythrocytes by G. Gardos (1958 BBA) but it is present in many cell types including pancreas cells where it is called KCa3.1 or KCNN4 (Maher and Kuchel, 2003).
  • the locus of the gene encoding the Gardos channel (KCNN4 protein) is mapped 19ql3.2.
  • the Gardos channel is made of 4 identical subunits; each subunit is encoded by a single gene, KCNN4, and comprises 6 transmembrane domains and a pore region between the 5 th and the 6 th transmembrane domains (Maher and Kuchel, 2003). In steady state conditions, the Gardos channel is inactive.
  • PIEZOs are newly identified mechanically activated (MA) cation channels, which responds to a wide array of mechanical forces, including poking, stretching, and shear stress, and is essential for proper vascular development in mice (Nilius and Honore, 2012; Li et al., 2014; Ranade et al., 2014). These ion channels are proposed homotrimers, with each subunit encompassing 14 transmembrane domains (Ge J. et al., 2015). They are expressed in many cell types including human erythroid progenitor cells (Zarychanski R et al., 2004; Coste B et al., 2010; Coste B et al., 2012).
  • This channel is permeable to Na + , K + and divalent cations such as Ca 2+ (Gnanasambandam R et al., 2015; Gottlied PA et al., 2012).
  • PIEZOl was shown to contribute to cell volume homeostasis by controlling Ca uptake and a functional link between PIEZOl and KCNN4 has been proposed (Cahalan et al., 2015). The opening of PIEZOl leads to a calcium influx that in turns activates KCNN4 mediating a K + efflux accompanied by CI " and osmotically linked water effluxes.
  • PIEZOl was also shown to regulate ATP release from human red blood cells by increasing calcium influx (Cinar E. et al., 2015). Thus, PIEZOl appears as a major contributor to red blood cell response to mechanical stress by controlling calcium influx.
  • KCNN4 activation alone is responsible for the red blood cell dehydration. They further demonstrated that the blocking of KCNN4 by an inhibitor of the Gardos channel completely reverses the red blood cell dehydration associated with the mutation of the PIEZOl channel. Thus, KCNN4 appears as the major effector of red blood cell dehydration in HX even in case of PIEZOl mutation.
  • the present invention therefore provides an inhibitor of the Gardos channel (i.e.: KCNN4 protein) for use in the treatment of hereditary xerocytosis (HX) in a subject, which is the carrier of at least one mutation of the PIEZOl gene encoding a mutated PIEZOl channel.
  • HX hereditary xerocytosis
  • the mutated PIEZOl channel is a human PIEZOl channel having at least one mutation as compared to the wild- type human PIEZOl channel, said mutation being selected from V598M, F681S, G718S/R2488Q, G782S/R808Q, S1117L/A2020V, R1358P, A2003D, A2020T, T2127M, DELK2166-2169, M2225R, R2456H, E2496ELE and R2488Q, preferably said least one mutation is selected from V598M and F681S.
  • HX is difficult to diagnose because of a highly variable expression, ranging from the absence of clinical symptoms to lethal perinatal oedema, but the most frequent HX condition is moderately symptomatic hemolysis.
  • the only test for HX is ektacytometry, which is available in a very limited number of laboratories. The disease may be overlooked for years or decades and is sometimes confused with spherocytosis. The provision of a diagnostic test is therefore or high clinical relevance.
  • the inventors have identified two additional single point mutations of the human
  • PIEZOl gene namely c.l792G>A or c.2042T>C resulting respectively in the mutation V598M or F681S in the PIEZOl channel, which participate in HX physiopathology and lead to a gain of function of the PIEZOl channel.
  • the mutations V598M and F681S are located in the N-terminal part of the extracellular domain of the protein, supposed to sense membrane shear stress (Ge J. et al., 2015).
  • the present application therefore further provides an in vitro method for diagnosing the presence of, or the predisposition to, hereditary xerocytosis (HX) in a human subject, comprising the step of:
  • the present invention relates to an inhibitor of the Gardos channel (KCNN4 protein) for use in the treatment of hereditary xerocytosis (HX) in a subject, characterized in that the subject is the carrier of at least one mutation of the PIEZOl gene encoding a PIEZOl channel having at least one mutation.
  • said at least one mutation is a gain of function mutation.
  • the PIEZOl channel is a mechanically- activated no specific cation channel that links mechanical forces to biological signals. As mentioned previously, it is permeable to Na + , K + and divalent cations such as Ca 2+ and generates currents characterized by a linear current- voltage relationship that are sensitive to ruthenium red and gadolinium. PIEZOl opening leads to a calcium influx in the red blood cell.
  • the PIEZOl channel is from human origin.
  • the amino acid sequence of the wild- type (WT) PIEZOl channel is available under accession number Q92508 in the UniProtKB database, and referred herein as SEQ ID NO: l.
  • the human Piezol channel is encoded by the gene PIEZOl .
  • the nucleic acid sequence of the wild-type gene (map: 16q24.3) encoding the human PIEZOl channel is available under ID number 9780 (NG_042229.1) in the NCBI GenBank database, and referred herein as SEQ ID NO:2.
  • the nucleic acid sequence of the mRNA (cDNA) encoded by the wild-type human PIEZOl gene is available under the accession number NM_001142864 in the NCBI GenBank database, and referred herein as SEQ ID NO:3.
  • the mutation in the PIEZOl gene may be any type of mutation, addition, deletion, missense mutation, or duplication. Typically the mutation is a missense mutation or a duplication.
  • gain of function mutation it is intended herein a mutation which produces an increased cation transport in erythroid cells. In a heterozygote, the new function will be expressed, and therefore the gain-of-function mutation will act like a dominant allele and produce the new phenotype mutated channel.
  • the subject is a human subject.
  • the mutated PIEZOl channel is a human PIEZOl channel having at least one mutation as compared to the WT human PIEZOl channel.
  • said at least one mutation is selected from V598M, F681S, G718S, R2488Q, G782S, R808Q, S1117L, A2020V, R1358P, A2003D, A2020T T2127M, DELK2166-2169, M2225R, R2456H, E2496ELE and R2488Q
  • optionally said at least one mutation is selected from V598M, F681S, G718S/R2488Q, G782S/R808Q, S1117L/A2020V, R1358P, A2003D, A2020T T2127M, DELK2166-2169, M2225R, R2456H, E2496ELE and R2488Q
  • said at least one mutation is selected from V598M and F681S.
  • V598M should be intended as an amino acid change from valine to methionine at position 598 of the WT human PIEZOl channel.
  • the same nomenclature applies to all other mutations mentioned in the present application.
  • the mutation G782S/R808Q should be intended as the double mutation G782S and R808Q.
  • E2496ELE means that the amino acid leucine and glutamate respectively in position 2495 and 2496 are duplicated. Said mutations are described in the documents of Zarychinski et al., 2012; Albuisson J et al., 2013 and Andolfo I et al., 2013, the content of which is included herein by reference. The single mutations V598M and F681S have now been discovered by the inventors.
  • the Gardos channel is a Ca 2+ sensitive, intermediate conductance, potassium selective channel also referred to as KCa3.1 or KCNN4 (Maher and Kuchel, 2003).
  • the Gardos channel is from human origin.
  • the amino acid sequence of the wild-type human Gardos channel (KCNN4 protein) is available under accession number 015554 (GI: 17366160) in the UniProtKB database, and referred herein to as SEQ ID NO: 4.
  • An inhibitor of the Gardos channel refers to a selective Gardos channel blocker that specifically inhibits the efflux of potassium from the erythrocytes.
  • An inhibitor of the Gardos channel can be identified by screening a collection of candidate compounds for their ability to specifically inhibit the efflux of potassium from the erythrocytes. Methods for measuring the inhibition of the efflux of potassium from the erythrocytes are known in themselves. Examples of such methods are described in Brugnara et al., 1993a and 1993b; Ellory et al., 1994. Both the percent inhibition of the Gardos channel and the 1 ⁇ 2 0 of an inhibitor of the Gardos channel can be assayed utilizing the methods described in Brugnara et al., 1993b.
  • the potency of an inhibitor of the Gardos channel can be assayed using erythrocytes by a method such as that disclosed by Brugnara et al., 1993a.
  • Inhibitors of the Gardos channel include organic molecules (e. g. a small organic molecule (natural or not)), amino acids, antibodies or aptamers.
  • Aptamers are a class of molecules that represents an alternative to antibodies in term of molecular recognition.
  • Aptamers are oligonucleotide or oligopeptide sequences with the capacity to recognize virtually any class of target molecules with high affinity and specificity.
  • Such ligands may be isolated through Systematic Evolution of Ligands by Exponential enrichment (SELEX) of a random sequence library, as described in Tuerk C. and Gold L., 1990.
  • the random sequence library is obtainable by combinatorial chemical synthesis of DNA. In this library, each member is a linear oligomer, eventually chemically modified, of a unique sequence. Possible modifications, uses and advantages of this class of molecules have been reviewed in Jayasena S.D., 1999.
  • Peptide aptamers consist of conformationally constrained antibody variable regions displayed by a platform protein, such as E. coli Thioredoxin A that are selected from combinatorial libraries by two hybrid methods (Colas et al, 1996).
  • the antibodies can be polyclonal or monoclonal antibodies.
  • the term "antibody” or “antibodies” as used herein also encompasses functional fragments of antibodies, including fragments of chimeric, humanized, single chain antibodies or fragments thereof (e.g., antigen- binding fragments of antibodies that specifically bind human the Gardos channel, Fv, Fab, Fab'and F(ab') 2 fragments).
  • Suitable antibodies are those which are directed to KCNN4 protein (Gardos channel).
  • said antibody is a monoclonal antibody, or fragment thereof.
  • the inhibitor of the Gardos channel is selected from the group consisting of imidazole antimycotics (Brugnara et al., 1996), such as clotrimazole (Brugnara et al., 1993a) metronidazole (Brugnara et al., 1993a), econazole (Brugnara et al., 1993a); arginine (Romero et al., 2002); Tram-34 (l-[(2-Chlorophenyl)diphenylmethyl]-lH- pyrazole) (Wulff et al, 2000); Charybdo toxin; Maurotoxin (Castle et al., 2002); nifedipine (Brugnara et al , 1993a); Nitrendipine (Brugnara et al, 1993a); inhibitors of calcium activated potassium flux that display selectivity and a potency towards the Gardos channel
  • the inhibitor of the Gardos channel is senicapoc or TRAM-34.
  • the inhibitor of the Gardos channel can be administered by any suitable route, for example, intravenously, intranasally, peritoneally, intramuscularly, orally and other conventional methods.
  • Said inhibitor according to the invention can included in a composition. It can be mixed and/or carried with one or more liquid and/or solid pharmaceutically acceptable carriers, ingredients and/or excipients.
  • pharmaceutically acceptable carrier is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Supplementary active compounds can also be incorporated into the composition.
  • treatment includes the administration of an inhibitor of the Gardos channel as defined above to a subject who is the carrier of at least one mutation of the PIEZOl gene encoding the PIEZOl channel, with the purpose to alleviate, relieve, alter, remedy, ameliorate, improve or affect this disorder.
  • the present invention also provides a method for treating hereditary xerocytosis, in a subject who is the carrier of at least one mutation of the PIEZOl gene encoding the PIEZOl channel as previously identified.
  • This method of treatment comprises administering to said subject an effective amount of an inhibitor of the Gardos channel (KCNN4 protein) as defined above.
  • the inhibitor of the Gardos channel is senicapoc or TRAM-34.
  • the present invention also provides the use of an inhibitor of the Gardos channel (KCNN4 protein) as defined above for the preparation of a medicament for treating hereditary xerocytosis in a subject who is the carrier of at least one mutation of the PIEZOl gene encoding the PIEZOl channel as previously identified.
  • KCNN4 protein an inhibitor of the Gardos channel
  • the present invention also provides a method of diagnosing and treating hereditary xerocytosis in a subject, comprising the steps of:
  • step (iii) diagnosing the subject with a hereditary xerocytosis when the presence of a mutation as defined in step (ii) in the biological sample is detected.
  • the method comprises an additional step iv) when the subject is diagnosed with hereditary xerocytosis is diagnosed consisting in administering an effective amount of an inhibitor of the Gardos channel (KCNN4 protein).
  • said inhibitor is senicapoc, or TRAM-34 as defined previously.
  • step (ii) rather comprises the detection in the said biological sample of at least one mutation in the PIEZOl gene (map: 16q24.3) as compared to the wild-type PIEZOl gene, preferably at least one mutation in the human PIEZOl gene (map: 16q24.3) as compared to the wild-type human PIEZOl gene which is responsible for the mutation V598M or F681S in the corresponding PIEZOl channel.
  • step (ii) can consist in detecting in the biological sample the presence of a missense mutation selected from c. l792G>A mutation in exon 14 or c.2042T>C mutation in exon 16 in the PIEZOl gene.
  • the term "detecting the presence of the missense mutation c. l792G>A or c.2042T>C refers to detecting the presence of the mutation corresponding to mutation c. l792G>A or c.2042T>C in the PIEZOl gene, either in mRNA or genomic DNA from a nucleic acid sample obtained from the subject.
  • the presence of said mutation c.l792G>A or c.2042T>C in the PIEZOl gene or of the corresponding mutation V598M or F681S in the PIEZOl channel constitutes a marker of a hereditary xerocytosis or a predisposition to hereditary xerocytosis in said subject.
  • a biological sample can be for example blood, serum, lymph, or any biological tissue.
  • the biological sample may also be pretreated, for example, by homogenization, extraction, enzymatic and/or chemical treatments as commonly used in the field.
  • FIGURES are a diagrammatic representation of FIGURES.
  • Figure 1 Red blood cell osmotic fragility test using osmotic gradient ranging from 0.1% to 1% of NaCl solution for healthy volunteers (A) and informed patients carrying the V598M mutation (B), the F681S mutation (C), or the G782S/R808Q double mutation (D) in the PIEZOl at TO and after 18 hours incubation at 37°C in presence or absence of Senicapoc at 0.4 or 4 ⁇ .
  • Figure 2 Variation in the red blood cell Na + (A) and K + (B) contents measured in red blood cells from informed patients carrying the F681S mutation, the G782S/R808Q double mutation, the V598M mutation or from healthy volunteers at TO (control condition, black bar), or after 18h incubation with 10 ⁇ Tram-34 (white bar), or 4 ⁇ of Senicapoc (grey bar).
  • C-D Cell water content (expressed in g water per g of dry weight) in patient red blood cells carrying the double mutation G782S/R808G (C) or control red blood cells from an healthy volunteer (D) measured at TO (control condition, black bar) and after 18h incubation with Senicapoc 4 ⁇ (dark grey bar) or 0.4 ⁇ (light grey bar).
  • Figure 3 Electrical features of patient red blood cells with Piezol mutations.
  • B Time course of currents measured at -80 mV for control, control+15 ⁇ Yodal or patient with G282S/R808Q Piezol. Data are median from currents recorded in A).
  • KCNN4 appears as the major effector of red blood cell dehydration in HX characterized so far and Senicapoc seems a promising therapeutic strategy to treat this condition whatever the mutated ion channel, Piezol or KCNN4.
  • Osmotic resistance tests Fresh venous blood was obtained by venipuncture from informed patients and healthy volunteers in EDTA collecting tubes kept at 4°C and sent overnight to Nice. At reception (24h after withdrawn) blood was washed 4 times at room temperature in medium containing in mM: NaCl 147, KC1 5, MgCl 2 1, CaCl 2 1, Hepes 10, buffered with NaOH pH7.4 (320 mOsm). Red blood cell suspension (40% hematocrit) was then incubated at 37°C for 18 hours in presence or absence of 10 ⁇ TRAM-34 or Senicapoc at 0.4 or 4 ⁇ . An osmotic resistance test in hypotonic saline solutions was performed on blood at reception and after 18 hours' incubation at 37°C.
  • Red blood cell cation content and volume measurements Red blood cell suspension before and after osmotic resistance test was used to quantify intracellular Na + and K + contents as previously described.
  • washed red blood cell suspension was set to 30% hematocrit and 0.5 ⁇ ouabain (Sigma- Aldrich) was added.
  • 15 ⁇ of Yodal was added to cell suspension either containing 10 ⁇ TRAM-34 (Chemi, 10 ⁇ GsMTx4 (Smartox) or DMSO. Samples were collected 15, 30 and 60 minutes after Yodal addition and cell water, Na + and K + contents were measured.
  • Electrophysiology All patch-clamp experiments were performed with a PC-controlled
  • EPC 9 patch-clamp amplifier (HEKA, Lambrecht/Pfalz, Germany). Currents were acquired and analyzed with Pulse and Pulsefit softwares (HEKA). Whole cell configuration was used and hematocrit was set at 10%. Glass pipettes (Brand, Wertheim, Germany) were made on a horizontal pipette puller (P-97; Sutter Instrument Co.; Navato, CA) to give a final resistance ranging from 18 to 20 ⁇ . For whole cell experiments, the bath solution was in mM: NaCl 150, KC1 5, MgCl 2 1, CaCl 2 1, Hepes 10, buffered with NaOH pH7.4 (320 mOsm).
  • the intracellular solution was in mM: KC1 140, NaCl 5, MgCl 2 1, Hepes 10 pH 7.2 adjusted with NaOH, 0.5 ⁇ CaCl 2 .
  • Currents were measured at room temperature using a ramp protocol from -120 to +80 mV from a holding potential of -60 mV (sampling frequency 10 kHz; filtered 1 kHz) Red blood cell suspension at 10% hematocrit.
  • PIEZOl sequencing revealed two new missense mutations: a c.l792G>A mutation in exon 14, leading to pVal598Met (i.e.: V598M) (predicted as tolerated by SIFT, score 0.1, and disease causing by Mutation taster, p value 0.998) and substitution in patient 2 and a c.2042T>C mutation in exon 16 leading to p.Phe681Ser substitution (i.e.: F681S) (predicted as deleterious by SIFT, score 0), in patient 3. No mutation was identified in KCNN4. Interestingly, in these 3 cases, HX mutations are not in the C terminal part of the protein, where most gain of function Piezo mutation were identified.
  • Osmotic resistance curves were done on washed red blood cells in Ca 2+ containing medium after 18 hours incubation at 37°C.
  • red blood cell Na + and K + contents were measured at time zero, before running the 18h incubation at 37°C and at the end of the incubation when osmotic resistance tests were done.
  • Table 2 Cation movements in control red blood cells in presence of 15 ⁇ Yodal, activator of Piezol channel, with or without a KCNN4 blocker (TRAM-34 10 ⁇ ) or a Piezol blocker (GsMTx4 10 ⁇ ). Data are means + sem of 4 experiments. ** p ⁇ 0.001 comparison of control condition with the 3 conditions with Yodal. ++ p ⁇ 0.001 comparison between Yodal and Yodal +inhibitors. Mann and Whitney test.
  • Piezol activation by Yodal induced a rapid change in cation contents with a steady state reached in 15 minutes (data not shown).
  • the water loss following Piezol activation is very faint and only due to the subsequent activation of KCNN4 as it is not observed in presence of KCNN4 inhibitor Tram-34.
  • FIG. 3A illustrates the currents in RBCs with mutated Piezol.
  • Figure 3B illustrates the kinetic of currents recorded at -80 mV for WT erythrocytes with or without Yodal and erythrocytes with G782S/R808Q mutated Piezol.
  • the large linear current observed in patient RBCs declined with a slower kinetic than the conductance induced by Yodal. It was possible to record currents in patient RBCs or control RBCs with Yodal when the large conductance was turned down. V curves are plotted on figure 3C.
  • Yodal effect on mouse red blood cell hydration suggested that it activates a calcium uptake via Piezol independently of any mechanical stress (Cahalan SM. Et al., 2015).
  • Yodal is also able to induce a rapid change in human red blood cell Na + and K + contents. A steady state is reached within about 15 minutes for cation content and cell volume. The Na + uptake is counter-balanced by the K + leak in presence of KCNN4 inhibitors.
  • the Piezol toxin GsMTx4 is able to partially prevent red blood cell dehydration and Na + and K + movements.
  • the effect on dehydration is likely due to a partial inhibition of Ca 2+ uptake and hence a lower activation of KCNN4.
  • Metabolic differences could explain discrepancy between patients for intracellular Na + and K + levels, as well as the changes in Piezol conductance induced by the different mutations.
  • red blood cell dehydration occurring in blood flow each time Piezol is activated by cell stretching might be less severe than with a constitutively activated Piezol channel.
  • all the mutations described in HX generate an inappropriate Piezol activity that is responsible for the higher Gardos channel activity leading to red blood cell dehydration.
  • the ability of red blood cells to cope with a hyperactive Gardos channel will depend on many other issues such as metabolism and pump efficiency. Senicapoc has already been shown to prevent red blood cell dehydration due to gain of function mutations in Gardos channel (Rapetti-Mauss R. et ah, 2016), the other molecular cause of HX. Thus this drug could also be used to treat HX due to gain of function mutations in Piezol.
  • Grootenboer S Schischmanoff PO, Cynober T, et al., Br J Haematol. 1998;103(2):383-386. Grootenboer S, Schischmanoff P-O, Laurendeau I et al., Blood. 2000;96(7):2599-2605.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Diabetes (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Cell Biology (AREA)
  • Organic Chemistry (AREA)
  • Microbiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne le traitement et le diagnostic de la xérocytose héréditaire (XH). Les inventeurs ont démontré que dans le cas d'une XH due à des mutations de PIEZO1, l'activation de KCNN4 est responsable de la déshydratation des érythrocytes, et que ladite déshydratation est inversée par l'inhibition du canal Gardos (protéine KCNN4). Ainsi, la présente invention concerne un inhibiteur du canal Gardos destiné à être utilisé dans le traitement de XH chez un sujet qui porte au moins une mutation du gène PIEZO1. En particulier, les inventeurs ont démontré que le Sénicapoc est efficace dans la prévention de la déshydratation des érythrocytes du patient portant une mutation de PIEZO. Ils ont également identifié que les mutations V598M ou F681S du canal PIEZO1 conduisent à un gain de fonctionnalité du canal et participent à la physiopathologie de la XH. Ainsi, la présente invention concerne également une méthode in vitro du diagnostic de XH, comprenant la détection dans un échantillon biologique de la présence de l'une quelconque de ces mutations.
PCT/EP2018/055373 2017-03-06 2018-03-05 Traitement et diagnostic d'une xérocytose héréditaire WO2018162426A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP17305234 2017-03-06
EP17305234.1 2017-03-06
EP17305290 2017-03-16
EP17305290.3 2017-03-16

Publications (1)

Publication Number Publication Date
WO2018162426A1 true WO2018162426A1 (fr) 2018-09-13

Family

ID=63448360

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2018/055373 WO2018162426A1 (fr) 2017-03-06 2018-03-05 Traitement et diagnostic d'une xérocytose héréditaire

Country Status (1)

Country Link
WO (1) WO2018162426A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110411991A (zh) * 2019-05-22 2019-11-05 郑州大学 一种对活细胞中Piezo1蛋白的超声敏感性的鉴定系统及方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000050026A1 (fr) 1999-02-23 2000-08-31 Icagen, Inc. Antagonistes du canal de gardos
WO2004016221A2 (fr) 2002-08-15 2004-02-26 Icagen, Inc Sulfonamides utilises comme agents de blocage des canaux a potassium
WO2005113490A1 (fr) 2004-05-21 2005-12-01 Icagen, Inc. Promedicaments contenant du sulfone
WO2006084031A1 (fr) 2005-02-01 2006-08-10 Icagen, Inc. Imines modulateurs du canal ionique
WO2014064465A1 (fr) * 2012-10-25 2014-05-01 Nhs Blood And Transplant Procédé pour détecter des fuites de cations
EP3106155A1 (fr) * 2015-06-15 2016-12-21 Universite d'Aix Marseille Traitement et diagnostic de la xérocytose héréditaire

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000050026A1 (fr) 1999-02-23 2000-08-31 Icagen, Inc. Antagonistes du canal de gardos
WO2004016221A2 (fr) 2002-08-15 2004-02-26 Icagen, Inc Sulfonamides utilises comme agents de blocage des canaux a potassium
WO2005113490A1 (fr) 2004-05-21 2005-12-01 Icagen, Inc. Promedicaments contenant du sulfone
WO2006084031A1 (fr) 2005-02-01 2006-08-10 Icagen, Inc. Imines modulateurs du canal ionique
WO2014064465A1 (fr) * 2012-10-25 2014-05-01 Nhs Blood And Transplant Procédé pour détecter des fuites de cations
EP3106155A1 (fr) * 2015-06-15 2016-12-21 Universite d'Aix Marseille Traitement et diagnostic de la xérocytose héréditaire
WO2016202854A1 (fr) 2015-06-15 2016-12-22 Universite D'aix-Marseille Traitement et diagnostic d'une xérocytose héréditaire

Non-Patent Citations (70)

* Cited by examiner, † Cited by third party
Title
ALBUISSON J; MURTHY SE; BANDELL M ET AL., NAT COMMUN., vol. 4, 2013, pages 1884
ANDOLFO I ET AL., AM J HEMATOL, vol. 88, 2013, pages 66 - 72
ANDOLFO I ET AL., AM J HEMATOL, vol. 90, 2015, pages 921 - 926
ANDOLFO I ET AL., BLOOD, vol. 121, 2013, pages 3925 - 3935
ARCHER NM ET AL., AM J HEMATOL, vol. 89, no. 1, 2014, pages 142 - 1146
ATAGA KI ET AL., BLOOD, vol. 11 1, 2008, pages 3991 - 3997
ATAGA KI ET AL., BR J HAEMATOL, vol. 153, 2011, pages 92 - 104
ATAGA KI ET AL., EXPERT OPIN INVESTIG DRUGS, vol. 18, 2009, pages 231 - 239
ATAGA KI; STOCKER J, EXPERT OPIN INVESTIG DRUGS, vol. 18, no. 2, 2009, pages 231 - 239
BAE C ET AL., PROC NATL ACAD SCI USA, vol. 1 10, 2013, pages E1 162 - 1 168
BAE C; GOTTLIEB PA; SACHS F, BIOPHYS J., vol. 105, no. 4, 2013, pages 880 - 886
BAE C; SACHS F; GOTTLIEB PA, BIOCHEMISTRY, vol. 50, no. 29, 2011, pages 6295 - 6300
BARNEAUD-ROCCA ET AL., J BIOL. CHEM., vol. 286, 2011, pages 8909 - 8916
BEAURAIN G; MATHIEU F; GROOTENBOER S ET AL., EUROPEAN JOURNAL OF HAEMATOLOGY, vol. 78, no. 3, 2007, pages 253 - 259
BENNEKOU P ET AL., BLOOD CELLS MOL. DIS., vol. 48, 2012, pages 102 - 109
BRUCE LJ ET AL., NAT GENET, vol. 37, 2005, pages 1258 - 1263
BRUGNARA C ET AL., J CLIN INVEST, vol. 92, 1993, pages 520 - 526
BRUGNARA C ET AL., J CLIN INVEST, vol. 97, 1996, pages 1227 - 1234
BRUGNARA C ET AL., J. BIOL. CHEM., vol. 268, 1993, pages 8760 - 8768
CAHALAN SM ET AL., ELIFE, 2015, pages 4
CARELLA M ET AL., AM J HUM GENET, vol. 63, 1998, pages 810 - 816
CARLI P ET AL., LA REVUE DE MEDECINE INTERNE, vol. 28, no. 12, 2007, pages 879 - 881
CASTLE A ET AL., MOL. PHARMACOL., vol. 63, 2002, pages 409 - 18
CINAR E ET AL., PROC NATL ACAD SCI USA, vol. 112, no. 38, 2015, pages 11783 - 11788
COSTE B ET AL., SCIENCE, vol. 330, 2010, pages 55 - 60
COSTE B; XIAO B; SANTOS JS ET AL., NATURE, vol. 483, no. 7388, 2012, pages 176 - 181
DA COSTA L ET AL., BLOOD REV., vol. 27, 2013, pages 167 - 178
DELAUNAY J., SEMIN HEMATOL, vol. 41, no. 2, April 2004 (2004-04-01), pages 165 - 72
DYRDA A ET AL., PLOS ONE, vol. 5, 2010, pages e9447
ELLORY JC ET AL., BR J PHARMACOL, vol. 1 1 1, 1994, pages 903 - 905
FANGER CM ET AL., J BIOL CHEM., vol. 274, 1999, pages 5746 - 5754
GE J; LI W; ZHAO Q ET AL., NATURE, vol. 527, no. 7576, 2015, pages 64 - 69
GLADER BE ET AL., N ENGL J MED., vol. 291, no. 10, 1974, pages 491 - 496
GLOGOWSKA E ET AL., BLOOD, vol. 126, 2015, pages 1281 - 1284
GNANASAMBANDAM R; BAE C; GOTTLIEB PA; SACHS F, PLOS ONE, vol. 10, no. 5, 2015, pages e0125503
GOTTLIEB PA; SACHS F, CHANNELS (AUSTIN, vol. 6, no. 4, 2012, pages 214 - 219
GROOTENBOER S; SCHISCHMANOFF PO; CYNOBER T ET AL., BR J HAEMATOL, vol. 103, no. 2, 1998, pages 383 - 386
GROOTENBOER S; SCHISCHMANOFF P-O; LAURENDEAU I ET AL., BLOOD, vol. 96, no. 7, 2000, pages 2599 - 2605
HOUSTON BL ET AL., BLOOD CELLS MOL DIS., vol. 47, 2011, pages 226 - 231
I. ANDOLFO ET AL: "Multiple clinical forms of dehydrated hereditary stomatocytosis arise from mutations in PIEZO1", BLOOD, vol. 121, no. 19, 11 March 2013 (2013-03-11), pages 3925 - 3935, XP055092445, ISSN: 0006-4971, DOI: 10.1182/blood-2013-02-482489 *
JOINER WJ ET AL., J BIOL CHEM., vol. 276, 2001, pages 37980 - 5
JOINER WJ ET AL., PROC NATL ACAD SCI USA, vol. 94, 1997, pages 11013 - 11018
KAPLAN JM; DELPECH M.: "Biologie Moleculaire et Medecine", 2007
LANG F ET AL., ADV EXP MED BIOL., vol. 559, 2004, pages 21 1 - 217
LI J ET AL., NATURE, vol. 515, 2014, pages 279 - 282
MAHER AD; KUCHEL PW, CELL BIOL., vol. 35, no. 8, 2003, pages 1182 - 1197
MARTIAL S ET AL., J CELL PHYSIOL., vol. 213, 2007, pages 70 - 78
MARTINAUD C; GISSEROT O; GRAFFIN B ET AL., THROMBOSIS RESEARCH., vol. 122, no. 4, 2008, pages 572 - 575
MILLER DR ET AL., BLOOD, vol. 38, no. 2, 1971, pages 184 - 204
MORALES P ET AL., J GEN PHYSIOL., vol. 142, 2013, pages 37 - 60
NILIUS B; E HONORE, TRENDS IN NEUROSCIENCES, vol. 35, 2012, pages 477 - 486
QUINLAN AR; HALL IM, BIOINFORMATICS, vol. 26, 2010, pages 841 - 842
RANADE SS ET AL., PNAS, vol. 111, 2014, pages 10347 - 10352
RAPETTI-MAUSS R ET AL., BLOOD, vol. 126, 2015, pages 1273 - 1280
RAPETTI-MAUSS R ET AL., HAEMATOLOGICA, vol. 101, no. 11, 2016, pages e431 - e435
RAPHAËL RAPETTI-MAUSS ET AL: "Red blood cell Gardos channel (KCNN4): the essential determinant of erythrocyte dehydration in hereditary xerocytosis", HAEMATOLOGICA, THE HEMATOLOGY JOURNAL : OFFICIAL ORGAN OF THE EUROPEAN HEMATOLOGY ASSOCIATION, vol. 102, no. 10, 15 June 2017 (2017-06-15), IT, pages e415 - e418, XP055472776, ISSN: 0390-6078, DOI: 10.3324/haematol.2017.171389 *
RINEHART J ET AL., CURR OPIN HEMATOL, vol. 17, 2010, pages 191 - 197
ROMERO JR ET AL., BLOOD, vol. 99, 2002, pages 1 103 - 1 108
STOCKER JW ET AL., BLOOD, vol. 101, no. 6, 2003, pages 2412 - 2418
SUCHYNA TM; JOHNSON JH; HAMER K ET AL., J GEN PHYSIOL., vol. 115, no. 5, 2000, pages 583 - 598
SYEDA R; XU J; DUBIN AE ET AL., ELIFE, 2015, pages 4
SYFUSS PY ET AL., CLINICAL & LABORATORY HAEMATOLOGY, vol. 28, no. 4, 2006, pages 270 - 274
VARECKA L; CARAFOLI E., J BIOL CHEM., vol. 257, 1982, pages 7414 - 7421
WANG K ET AL., NUCLEIC ACIDS RES., vol. 38, 2010, pages el 64
WEISS E ET AL., ANEMIA, 2011, pages 379894
WU J; LEWIS AH; GRANDL J, TRENDS BIOCHEM SCI., vol. 42, no. 1, 2017, pages 57 - 71
WULFF H ET AL., PNAS, vol. 97, 2000, pages 8151 - 8156
WULFF H; CASTLE NA, EXPERT REV CLIN PHARMACOL, vol. 3, 2010, pages 385 - 396
WULFF H; KOHLER R, J CARDIOVASC PHARMACOL, vol. 61, 2013, pages 102 - 112
ZARYCHANSKI R ET AL., BLOOD, vol. 120, 2012, pages 1908 - 1915

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110411991A (zh) * 2019-05-22 2019-11-05 郑州大学 一种对活细胞中Piezo1蛋白的超声敏感性的鉴定系统及方法

Similar Documents

Publication Publication Date Title
Rapetti-Mauss et al. A mutation in the Gardos channel is associated with hereditary xerocytosis
Jenkins et al. Lipoproteins, glycoxidation and diabetic angiopathy
Tan et al. Association between serum levels of soluble receptor for advanced glycation end products and circulating advanced glycation end products in type 2 diabetes
Helmond et al. Treatment of immune‐mediated hemolytic anemia with individually adjusted heparin dosing in dogs
Ferru et al. Thalassemic erythrocytes release microparticles loaded with hemichromes by redox activation of p72Syk kinase
Koizumi et al. Therapeutic effects of ulinastatin on experimental crescentic glomerulonephritis in rats
Bissinger et al. Proteinuric chronic kidney disease is associated with altered red blood cell lifespan, deformability and metabolism
US20140179700A1 (en) Methods of treating hemoglobinopathies
Caulier et al. Primary red cell hydration disorders: pathogenesis and diagnosis
Kamal et al. New emerging roles of the novel hepatokine SERPINB1 in type 2 diabetes mellitus: Crosstalk with β-cell dysfunction and dyslipidemia
WO2015179855A1 (fr) Procédés pour limiter la morbidité dans les cas d&#39;hémoglobinopathie
WO2018162426A1 (fr) Traitement et diagnostic d&#39;une xérocytose héréditaire
KR20090083335A (ko) 간 기능 평가용 바이오마커
Hu et al. The critical role of pancreatic stone protein/regenerating protein in sepsis-related multiorgan failure
Murakoshi et al. Pleiotropic effect of pyridoxamine on diabetic complications via CD36 expression in KK-Ay/Ta mice
Nagano et al. Expression of a Crry/p65 is reduced in acute lung injury induced by extracellular histones
Agouti et al. Plasma levels of E‐selectin are associated with retinopathy in sickle cell disease
US20180136227A1 (en) Treatment and Diagnosis of Hereditary Xerocytosis
Hirata et al. Serum lipoprotein (a) concentrations and apolipoprotein (a) phenotypes in the families of NIDDM patients
Dong et al. Fibrinogen-like protein 2 prothrombinase may contribute to the progression of inflammatory bowel disease by mediating immune coagulation
Tan et al. Molecular basis of kidney disease
EP3655014A1 (fr) Clusterine pour son utilisation dans le traitement des micro-angiopathies thrombotiques
Smith et al. Platelet noradrenaline release in patients with familial hypercholesterolaemia
Hamdan et al. The Diagnostic Value of Serum Ferritin in Assessing Iron Status in Syrian Hemodialysis Patients
Kang et al. Aliskiren regulates neonatal Fc receptor and IgG metabolism with attenuation of anti-GBM glomerulonephritis in mice

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18712516

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18712516

Country of ref document: EP

Kind code of ref document: A1