Methods for preventing or treating disorders by increasing bioavailability of iron and related pharmaceutical formulation

Abstract

The present disclosure relates to methods of treating, ameliorating or preventing a disorder comprising administering a therapeutically effective amount of a composition to a subject in need thereof, which composition contains a lipocalin mutein or a fragment or a variant thereof capable of increasing the bioavailability of iron in the subject.

1 . A method of treating, ameliorating or preventing a disorder associated with an altered level of iron, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a lipocalin mutein or a fragment thereof having binding affinity to hepcidin,
 wherein the disorder associated with an altered level of iron is anemia,   wherein the lipocalin mutein has the same amino acids as the mutein set forth in SEQ ID NO: 1 at two or more positions corresponding to positions 36, 40, 41, 49, 52, 68, 70, 72, 73, 77, 79, 81, 96, 100, 103, 106, 125, 127, 132, and 134 of the linear polypeptide sequence of the mature human neutrophil gelatinase-associated lipocalin (hNGAL).   
     
     
         2 . The method of  claim 1 , wherein said pharmaceutical composition is administered by intracutaneous, subcutaneous, intramuscular or intravenous injection or by infusion techniques. 
     
     
         3 . The method of  claim 1 , wherein said pharmaceutical composition is administered via an enteral route. 
     
     
         4 . The method of  claim 1 , wherein said pharmaceutical composition is administered up to twice daily, up to once daily, up to once every other day, up to once every third day, up to twice every week, up to once every week, up to once every other week, or up to once every month. 
     
     
         5 . The method of  claim 1 , wherein said pharmaceutical composition is administered to a subject in need thereof, each time at a dosage level selected from the group consisting of: 0.1-1 mg/kg, 0.1-40 mg/kg, 1-20 mg/kg, 1-10 mg/kg, 3-20 mg/kg and 1-3 mg/kg. 
     
     
         6 . The method of  claim 1 , wherein said pharmaceutical composition is administered to a subject in need thereof, each time at a dosage level of 3 mg/kg to 10 mg/kg. 
     
     
         7 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof is capable of increasing the bioavailability of iron in the subject. 
     
     
         8 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof is capable of inhibiting binding of hepcidin to a hepcidin specific monoclonal antibody having the variable light chain region shown in SEQ ID NO: 6 and the variable heavy chain region shown in SEQ ID NO: 7. 
     
     
         9 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof competes for binding to hepcidin with a hepcidin specific monoclonal antibody having the variable light chain region shown in SEQ ID NO: 6 and the variable heavy chain region shown in SEQ ID NO: 7. 
     
     
         10 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof has the same amino acids as the mutein set forth in SEQ ID NO: 1 at the positions corresponding to positions 36, 40, 41, 49, 52, 68, 70, 72, 73, 77, 79, 81, 96, 100, 103, 106, 125, 127, 132, and 134 of the linear polypeptide sequence of mature hNGAL. 
     
     
         11 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof has at least 80% sequence identity to SEQ ID NO: 1. 
     
     
         12 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof has at least 95% sequence identity to SEQ ID NO: 1. 
     
     
         13 . The method of  claim 1 , wherein said lipocalin mutein or a fragment thereof is conjugated to a compound that extends the serum half-life of the mutein. 
     
     
         14 . The method of  claim 13 , wherein said compound that extends the serum half-life of the mutein is selected from the group consisting of a polyalkylene glycol molecule, a hydroxy ethyl starch, a protein domain, a Fc part of an immunoglobulin, a CH3 domain of an immunoglobulin, a CH4 domain of an immunoglobulin, an albumin-binding peptide, and an albumin-binding protein. 
     
     
         15 . The method of  claim 14 , wherein said polyalkylene glycol is polyethylene glycol (PEG) or an activated derivative thereof, and wherein said polyethylene glycol (PEG) is preferably 30 kiloDalton in molecular weight. 
     
     
         16 . The method of  claim 1 , wherein said anemia is anemia of inflammation, chronic inflammatory anemia, an iron-deficiency anemia, an iron loading anemia, anemia associated with chronic kidney disease (CKD), anemia of cancer (AC), chemotherapy induced anemia (CIA), or an anemia associated with erythropoiesis-stimulating agent (ESA)-resistance. 
     
     
         17 . A concentrated, stable pharmaceutical formulation of at least one lipocalin mutein which is capable of increasing the bioavailability of iron in a subject in need thereof, wherein the lipocalin mutein has a sequence identity of at least 80% to the amino acid sequence of SEQ ID NO: 1, comprising:
 a. up to about to 350 mg/ml of the lipocalin mutein;   b. about 1 to 100 mM of a buffering agent providing a pH of 5.5 to 8;   c. about 1 to 500 mM of a stabilizer or a mixture of two or more stabilizers;   d. about 0.01 to 0.08% of a non-ionic surfactant; and   e. an effective amount of at least one hyaluronidase enzyme.   
     
     
         18 . The formulation according to  claim 17  for use in a method of treating a disorder which is amenable to treatment with a lipocalin mutein or a fragment thereof capable of increasing the bioavailability of iron in a subject in need thereof, comprising the step of administering to the subject an amount effective to treat the said disorder.