Targeted protein replacement for the treatment of lysosomal storage disorders
The present invention relates to compositions and methods for delivering lysosomal proteins. The compositions and methods described herein permit the targeted delivery of exogenous lysosomal proteins to cell surface proteins that allow their internalization via non-clathrin pathways. The present invention further relates to the use of the compositions and methods for enzyme replacement therapy of lysosomal storage diseases. Nucleic acids, recombinant cells and kits useful for making and using the compositions of the invention are also provided.
1. A composition comprising a mammalian acid sphingomyelinase or an active fragment thereof which is attached to a targeting moiety, wherein the targeting moiety binds to an extracellular portion of Intercellular Adhesion Molecule-1 (ICAM-1) or Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1).
2. The composition of claim 1 wherein the mammalian acid sphingomyelinase or active fragment thereof is covalently attached to said targeting moiety.
3. The composition of claim 2 wherein the mammalian acid sphingomyelinase or active fragment thereof is a fusion protein comprising said targeting moiety and said acid sphingomyelinase or said fragment.
4. The composition of claim 3 wherein the targeting moiety is N-terminal to said acid sphingomyelinase or said fragment thereof.
5. The composition of claim 3 wherein the targeting moiety is C-terminal to said acid sphingomyelinase or said fragment thereof.
6. The composition of claim 2 wherein the mammalian acid sphingomyelinase or active fragment thereof is conjugated to said targeting moiety.
7. The composition of claim 6 wherein the mammalian acid sphingomyelinase or active fragment thereof is conjugated to said targeting moiety by chemical cross-linking.
8. The composition of claim 1 wherein the targeting moiety is an antibody or a non-immunoglobulin polypeptide.
9. The composition of claim 8 wherein the targeting moiety is an antibody.
10. The composition of claim 1 , wherein the mammalian acid sphingomyelinase or active fragment thereof is at least 95% pure.
11. A particle comprising a mammalian acid sphingomyelinase or an active fragment thereof which is attached to a targeting moiety, wherein the targeting moiety binds to an extracellular portion of Intercellular Adhesion Molecule-1 (ICAM-1) or Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1).
12. The particle of claim 11 wherein the targeting moiety is an antibody or a non-immunoglobulin polypeptide.
13. The particle of claim 11 wherein the mammalian acid sphingomyelinase or active fragment thereof is at least 95% pure.
14. The particle of claim 11 which is 50 nm to 10 μm in size.
15. The particle of claim 14 which is 200-300 nm in size.
16. The particle of claim 11 which is a synthetic carrier particle.
17. The particle of claim 11 which is a liposome, a microbubble, a dendrimer, or a micelle.
18. The particle of claim 16 in which the synthetic carrier particle is coupled to, loaded into, loaded onto or coated with said targeting moiety.
19. A pharmaceutical composition comprising (a) the composition of claim 1 and (b) a pharmaceutically acceptable carrier.
20. A pharmaceutical composition comprising (a) the composition of claim 10 and (b) a pharmaceutically acceptable carrier.
21. A pharmaceutical composition comprising (a) the particle of claim 11 and (b) a pharmaceutically acceptable carrier.
22. A pharmaceutical composition comprising (a) the particle of claim 14 and (b) a pharmaceutically acceptable carrier.
23. A method for treating a subject in need of acid sphingomyelinase replacement therapy, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition of claim 19 .
24. A method for treating a subject in need of acid sphingomyelinase replacement therapy, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition of claim 20 .
25. A method for treating a subject in need of acid sphingomyelinase replacement therapy, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition of claim 21 .
26. A method for treating a subject in need of acid sphingomyelinase replacement therapy, said method comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition of claim 22 .
27. The method of claim 23 , wherein the subject has Niemann-Pick disease type A or Niemann-Pick disease type B.
28. The method of claim 24 , wherein the subject has Niemann-Pick disease type A or Niemann-Pick disease type B.
29. The method of claim 25 , wherein the subject has Niemann-Pick disease type A or Niemann-Pick disease type B.
30. The method of claim 26 , wherein the subject has Niemann-Pick disease type A or Niemann-Pick disease type B.
31. The composition of claim 1 , wherein the mammalian acid sphingomyelinase and the targeting moiety are absorbed onto the surface of a particle.
32. The composition of claim 31 , wherein the mammalian acid sphingomyelinase and the targeting moiety are absorbed onto the surface of the particle as separate molecules.
33. A mammalian acid sphingomyelinase or an active fragment thereof conjugated to an antibody that binds to Intercellular Adhesion Molecule-1 (ICAM-1).
34. A particle comprising a mammalian acid sphingomyelinase or an active fragment thereof, wherein the acid sphingomyelinase or active fragment thereof is coated on a particle and wherein an antibody that binds to Intercellular Adhesion Molecule-1 (ICAM-1) is also coated on the particle.
35. The composition of claim 32 , wherein the mammalian acid sphingomyelinase has acid sphingomyelinase activity.
36. The composition of claim 32 , wherein the particle is a latex bead.
37. The composition of claim 1 , wherein the mammalian acid sphingomyelinase is human acid sphingomyelinase.
38. The particle of claim 34 , wherein the mammalian acid sphingomyelinase is human acid sphingomyelinase.
39. The particle of claim 34 , wherein the mammalian acid sphingomyelinase has acid sphingomyelinase activity.
40. The particle of claim 34 , wherein the particle is a latex bead.