Methods for treating pompe disease

1 . A method for treating pompe disease in a subject comprising administering to the subject a therapeutically effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

2 . The method of claim 1 , wherein the lysosomal targeting domain comprises mature human insulin-like growth factor II (IGF-II) or a fragment or sequence variant thereof.

3 . The method of claim 2 , wherein the lysosomal targeting domain comprises amino acids 1 and 8-67 of mature human IGF-II.

4 . The method of claim 1 , wherein the fusion protein comprises amino acids 70-952 of human GAA.

5 . The method of claim 1 , wherein the fusion protein has reduced mannose-6-phosphate (M6P) level thereon compared to wild-type human GAA.

6 . The method of claim 1 , wherein the fusion protein has no functional M6P level thereon.

7 . The method of claim 1 , wherein the therapeutically effective amount is in the range of 2.5-20 mg per kilogram of body weight of the subject.

8 . The method of claim 1 , wherein the fusion protein is administered intravenously.

9 . The method of claim 1 , wherein the fusion protein is administered bimonthly, monthly, triweekly, biweekly, weekly, daily, or at variable intervals.

10 . A method for treating Pompe disease in a subject comprising administering to the subject a therapeutically effective amount of a fusion protein comprising amino acids 1 and 8-67 of mature human insulin-like growth factor II (IGF-II) and amino acids 70-952 of human acid alpha-glucosidase (GAA).

11 . The method of claim 10 , wherein the fusion protein further comprises a spacer sequence Gly-Ala-Pro between the amino acids of mature human IGF-II and the amino acids of human GAA.

12 . The method of claim 10 , wherein the fusion protein has reduced mannose-6-phosphate (M6P) level thereon compared to wild-type human GAA.

13 . The method of claim 10 , wherein the fusion protein has no functional M6P level thereon.

14 . A method for reducing glycogen levels in vivo comprising administering to a subject suffering from Pompe disease an effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

15 . The method of claim 14 , wherein the lysosomal targeting domain comprises mature human insulin-like growth factor II (IGF-II) or a fragment or sequence variant thereof.

16 . The method of claim 15 , wherein the lysosomal targeting domain comprises amino acids 1 and 8-67 of mature human IGF-II.

17 . The method of claim 14 , wherein the fusion protein comprises amino acids 70-952 of human GAA.

18 . The method of claim 14 , wherein the fusion protein has reduced mannose-6-phosphate (M6P) level thereon compared to wild-type human GAA.

19 . The method of claim 14 , wherein the fusion protein has no functional M6P level thereon.

20 . The method of claim 14 , wherein the therapeutically effective amount is in the range of 2.5-20 mg per kilogram of body weight of the subject.

21 . The method of claim 14 , wherein the fusion protein is administered intravenously.

22 . The method of claim 14 , wherein the fusion protein is administered bimonthly, monthly, triweekly, biweekly, weekly, daily, or at variable intervals.

23 . A method for reducing glycogen levels in a mammalian lysosome comprising targeting to the lysosome an effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

24 . The method of claim 23 , wherein the lysosomal targeting domain comprises mature human insulin-like growth factor II (IGF-II) or a fragment or sequence variant thereof.

25 . The method of claim 23 , wherein the lysosomal targeting domain comprises amino acids 1 and 8-67 of mature human IGF-II.

26 . The method of claim 23 , wherein the fusion protein comprises amino acids 70-952 of human GAA.

27 . A method for reducing glycogen levels in a muscle tissue of a subject suffering from Pompe disease comprising delivering to the muscle tissue a therapeutically effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

28 . The method of claim 27 , wherein the muscle tissue is skeletal muscle.

29 . A method for treating cardiomyopathy associated with Pompe disease in a subject comprising administering to the subject a therapeutically effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

30 . A method for treating myopathy associated with Pompe disease in a subject comprising administering to the subject a therapeutically effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

31 . A method for increasing acid alpha-glucosidase (GAA) activity in a subject suffering from Pompe disease comprising administering to the subject a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

32 . A pharmaceutical composition suitable for treatment of Pompe disease comprising a therapeutically effective amount of a fusion protein comprising human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain, wherein the lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

33 . The pharmaceutical composition of claim 32 , wherein the lysosomal targeting domain comprises mature human insulin-like growth factor II (IGF-II) or a fragment or sequence variant thereof.

34 . The pharmaceutical composition of claim 32 , wherein the lysosomal targeting domain comprises amino acids 1 and 8-67 of mature human IGF-II.

35 . The pharmaceutical composition of claim 32 , wherein the fusion protein comprises amino acids 70-952 of human GAA.

36 . The pharmaceutical composition of claim 32 , wherein the fusion protein comprises amino acids 70-952 of human GAA and amino acids 1 and 8-67 of mature human IGF-II.

37 . The pharmaceutical composition of claim 36 , wherein the fusion protein further comprises a spacer sequence Gly-Ala-Pro between the amino acids of human GAA and the amino acids of mature human IGF-II.

38 . The pharmaceutical composition of claim 32 , wherein the fusion protein has reduced mannose-6-phosphate (M6P) level thereon compared to wild-type human GAA.

39 . The pharmaceutical composition of claim 32 , wherein the fusion protein has no functional M6P level thereon.

40 . The pharmaceutical composition of claim 32 , wherein the pharmaceutical composition further comprises a pharmaceutical carrier.