Recombinant alpha-L-iduronidase, methods for producing and purifying the same and methods for treating diseases caused by deficiencies thereof

Abstract

The present invention provides a recombinant α-L-iduronidase and biologically active fragments and mutants thereof, methods to produce and purify this enzyme as well as methods to treat certain genetic disorders including—α-L-iduronidase deficiency and mucopolysaccharidosis I (MPS 1).

1 - 20 . (canceled)  
     
     
         21 . A method of treating diseases caused all or in part by a deficiency in α-L-iduronidase, comprising the steps of administering to a subject a pharmaceutical composition comprising a therapeutically effective amount of a recombinant α-L-iduronidase enzyme with a purity of equal to or greater than 99%, wherein said recombinant α-L-iduronidase enzyme comprises (a) the amino acid sequence of residues 26 to 653 of SEQ ID NO:2, or a biologically active fragment of SEQ ID NO:2 which retains α-L-iduronidase enzyme biological activity; and (b) one or more mannose-6-phosphate residues.  
     
     
         22 . The method of  claim 21  wherein the disease is mucopolysaccharidosis.  
     
     
         23 . The method of  claim 21  wherein the disease is mucopolysaccharidosis I.  
     
     
         24 . The method of  claim 21  wherein the disease is selected from the group consisting of Hurler's disease, Scheie syndrome and Hurler-Scheie syndrome.  
     
     
         25 . The method of  claim 21  wherein said subject suffering from the disease demonstrates about 1% or less of a normal α-L-iduronidase activity.  
     
     
         26 . The method of  claim 21  wherein a dose of at least about 125,000 units or 0.5 mg/kg of said human recombinant α-L-iduronidase is administered weekly to a patient suffering from a deficiency thereof.  
     
     
         27 . The method of  claim 21  wherein said administering is the slow infusion of at least 0.5 mg/kg of said formulation for about an hour, followed by a rapid two-hour infusion rate.  
     
     
         28 . The method of  claim 27  wherein said infusion is used to minimize complement mediation clinical allergic reactions.  
     
     
         29 . The method of  claim 21  wherein said treatment with human recombinant α-L-iduronidase reduces lysosomal storage caused all or in part by said deficiency in α-L-iduronidase of said subject.  
     
     
         30 . The method of  claim 21  wherein said treatment causes reduction in clinical and biochemical symptoms caused all or in part by said deficiency in α-L-iduronidase of said subject.  
     
     
         31 . The method of  claim 21  wherein said treatment results in normalization of liver volume and urinary glycosaminoglycan excretion, reduction in spleen size and apnea/hypopnea events, increase in height and growth velocity in prepubertal patients, increase in shoulder flexion and elbow and knee extension, and reduction in tricuspid regurgitation or pulmonic regurgitation.  
     
     
         32 . The method of  claim 21  wherein the recombinant α-L-iduronidase enzyme has a specific activity greater than 200,000 units per milligram protein.  
     
     
         33 . The method of  claim 21  wherein the recombinant α-L-iduronidase enzyme of has a specific activity greater than 240,000 units per milligram protein.  
     
     
         34 . The method of  claim 21  wherein the recombinant α-L-iduronidase enzyme is a recombinant human α-L-iduronidase enzyme.  
     
     
         35 . A method of treating diseases caused all or in part by a deficiency in α-L-iduronidase, comprising the steps of administering to a subject a pharmaceutical composition comprising a therapeutically effective amount of a recombinant α-L-iduronidase enzyme with a purity of equal to or greater than 99%, wherein said recombinant α-L-iduronidase enzyme comprises (a) the amino acid sequence of residues 26 to 653 of SEQ ID NO:2, or a biologically active fragment of SEQ ID NO:2 which retains α-L-iduronidase enzyme biological activity; and (b) has a specific activity greater than 200,000 units per milligram protein.  
     
     
         36 . The method of  claim 35  wherein the recombinant α-L-iduronidase enzyme comprises a mannose-6-phosphate residue attached at position 3 and a mannose-6-phosphate residue attached at position 6.  
     
     
         37 . The method of  claim 35  wherein the disease is mucopolysaccharidosis.  
     
     
         38 . The method of  claim 35  wherein the disease is mucopolysaccharidosis I.  
     
     
         39 . The method of  claim 35  wherein the disease is selected from the group consisting of: Hurler's disease, Scheie syndrome and Hurler-Scheie syndrome.  
     
     
         40 . The method of  claim 35  wherein said subject suffering from the disease demonstrates about 1% or less of a normal α-L-iduronidase activity.  
     
     
         41 . The method of  claim 35  wherein a dose of at least about 125,000 units or 0.5 mg/kg of said recombinant α-L-iduronidase is administered weekly to a patient suffering from a deficiency thereof.  
     
     
         42 . The method of  claim 35  wherein said administering is the slow infusion of at least 0.5 mg/kg of said formulation for about an hour, followed by a rapid two-hour infusion rate.  
     
     
         43 . The method of  claim 42  wherein said infusion is used to minimize complement mediation clinical allergic reactions.  
     
     
         44 . The method of  claim 35  wherein said treatment with human recombinant α-L-iduronidase reduces lysosomal storage caused all or in part by said deficiency in α-L-iduronidase of said subject.  
     
     
         45 . The method of  claim 35  wherein said treatment causes reduction in clinical and biochemical symptoms caused all or in part by said deficiency in α-L-iduronidase of said subject.  
     
     
         46 . The method of  claim 35  wherein said treatment results in normalization of liver volume and urinary glycosaminoglycan excretion, reduction in spleen size and apnea/hypopnea events, increase in height and growth velocity in prepubertal patients, increase in shoulder flexion and elbow and knee extension, and reduction in tricuspid regurgitation or pulmonic regurgitation.  
     
     
         47 . The method of  claim 35  wherein the recombinant α-L-iduronidase enzyme of has a specific activity greater than 240,000 units per milligram protein.  
     
     
         48 . The method of  claim 35  wherein the recombinant α-L-iduronidase enzyme is a recombinant human α-L-iduronidase enzyme.