Highly Potent Acid Alpha-Glucosidase With Enhanced Carbohydrates
Recombinant human alpha glucosidase (rhGAA) composition derived from CHO cells that contains a more optimized glycan composition consisting of a higher amount of rhGAA containing N-glycans carrying mannose-6-phosphate (M6P) or bis-M6P than conventional rhGAAs, along with low amount of non-phosphorylated high mannose glycans, and low amount of terminal galactose on complex oligosaccharides. Compositions containing the rhGAA, and methods of use are described
1 . A rhGAA composition derived from CHO cells that contains a higher amount of rhGAA containing N-glycans carrying mono-mannose-6-phosphate (M6P) or bis-M6P than conventional rhGAA as exemplified by Lumizyme® (Alglucosidase Alfa; CAS 420794-05-0).
2 . The rhGAA composition of claim 1 , wherein the rhGAA comprises an amino acid sequence that is at least 95% identical to that described by SEQ ID NO: 1.
3 . The rhGAA composition of claim 1 , wherein 30% or more of the rhGAA binds to cation-independent manose-6-phosphate receptor (CIMPR) or 30% or more of the rhGAA contains an N-glycan carrying mono-M6P or bis-M6P.
4 . The rhGAA composition of claim 1 , wherein 50% or more of the rhGAA binds to cation-independent manose-6-phosphate receptor (CIMPR) or 50% or more of the rhGAA contains an N-glycan carrying mono-M6P or bis-M6P.
5 . The rhGAA composition of claim 1 , wherein 90% to 100% of the rhGAA binds to cation-independent manose-6-phosphate receptor (CIMPR) or 90% to 100% or more of the rhGAA contains an N-glycan carrying mono-M6P or bis-M6P.
6 . The rhGAA composition of claim 1 wherein the average content of N-glycans carrying M6P ranges from 0.5 to 7.0 mol/mol rhGAA.
7 . The rhGAA composition of claim 1 wherein the average content of N-glycans carrying M6P ranges from 1.0 to 4.0 mol/mol rhGAA.
8 . The rhGAA composition of claim 1 wherein the average content of N-glycans carrying M6P ranges from 5.0 to 6.0 mol/mol rhGAA.
9 . The rhGAA composition of claim 1 , wherein on average the N-glycans contain greater than 3 mol/mol of M6P and greater than 4 mol/mol sialic acid.
10 . The rhGAA composition of claim 1 , wherein on average 40 to 60% of the N-glycans on the rhGAA are complex type N-glycans; wherein no more than 6.5% of N-glycans on the rhGAA are hybrid-type N-glycans, and wherein no more than 15% of the high mannose-type N-glycans on the rhGAA are non-phosphorylated, at least 10% of the high mannose-type N-glycans on the rhGAA are mono-M6P phosphorylated, and/or at least 2% of the high mannose-type N-glycans on the rhGAA are bis-M6P phosphorylated.
11 . The composition of claim 1 , wherein the rhGAA on average carries 2.0 to 8.0 sialic acid residues per mol of rhGAA.
12 . The composition of claim 1 , wherein the rhGAA is produced by CHO cell line ATB200-001-X5-14 or by a subculture thereof.
13 . The composition of claim 1 , further comprising a chaperone.
14 . A method for making the composition of claim 1 comprising culturing a CHO cell line and recovering said composition from the culture of CHO cells.
15 . A CHO cell line that produces the composition of claim 1 .
16 . CHO cell line GA-ATB-200 or a subculture thereof that produces the composition of claim 1 .
17 . A method for making the CHO cell line of claim 15 , comprising transforming a CHO cell with DNA encoding rhGAA, selecting a CHO cell that stably integrates the DNA encoding rhGAA into its chromosome(s) and that stably expresses rhGAA, and selecting a CHO cell that expresses rhGAA having a high content of glycans bearing M6P or bis-M6P, and, optionally, selecting a CHO cell having N-glycans with high sialic acid content and and/or having N-glycans with a low non-phosphorylated high-mannose content.
18 . A method for treating a condition, disorder or disease associated with insufficient lysosomal rhGAA comprising administering the composition of claim 1 to a subject in need thereof.
19 . The method of claim 18 , wherein the subject has Glycogen Storage Disease Type II (Pompe Disease).
20 . The method of claim 18 , comprising administering said composition to cardiac muscle of the subject.
21 - 27 . (canceled)