Targeted therapeutic lysosomal enzyme fusion proteins and uses thereof
The present invention relates in general to therapeutic fusion proteins useful to treat lysosomal storage diseases and methods for treating such diseases. Exemplary therapeutic fusion proteins comprise a lysosomal enzyme, a lysosomal targeting moiety, e.g., an IGF-II peptide, and a spacer peptide. Also provided are compositions and methods for treating Mucopolysaccharidosis Type IIIB (Sanfilippo B Syndrome), comprising a targeted therapeutic fusion protein comprising alpha-N-acetylglucosaminidase (Naglu), a lysosomal targeting moiety, e.g., an IGF-II peptide, and a spacer peptide.
1. A targeted therapeutic fusion protein comprising a human α-N-acetylglucosaminidase (Naglu) protein comprising amino acids 1-743 or 24-743 of SEQ ID NO: 1, an IGF-II peptide tag having an amino acid sequence at least 70% identical to amino acids 8-67 of SEQ ID NO: 5 and a spacer peptide located between the Naglu protein and the IGF-II peptide tag, wherein the spacer peptide comprises the amino acid sequence selected from the group consisting of:
GAPGGGGSGGGGSGGGGSGGGGSGGGPSGGGGSGGGPSGAP (SEQ ID NO: 29), GAPGGGGSGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGGSGGGGSGGGPSG AP (SEQ ID NO: 31), GAPGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 33), GAPGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSG AP (SEQ ID NO: 35), GAPGGGGSGGGGSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 37), GAPGGGGSGGGGSAAAASGGGGSGGGPSGAP (SEQ ID NO: 39), GAPGGGGSGGGGSAAAASGGGGSGGGGSAAAASGGGGSGGGGSAAAASGGGPS GAP (SEQ ID NO: 41), GAPGGGGSGGGGSAAAASGGGPSGGGGSAAAASGGGPSGGGGSAAAASGGGPSG AP (SEQ ID NO: 43), GAPGGGSPAPTPTPAPTPAPTPAGGGPSGAP (SEQ ID NO: 49), GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51), GAPGGGSPAEAAAKEAAAKEAAAKEAAAKEAAAKAPSGGGGAP (SEQ ID NO: 55), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGGGGAGGGPSGAP (SEQ ID NO: 64), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGGAGGGGAGGG PSGA P (SEQ ID NO:66), GAPGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGPSGAP (SEQ ID NO: 68), GAPGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGPSGGGGAGGGGAGGGP S GA P (SEQ ID NO: 70), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGAP (SEQ ID NO: 72), GAPGGGGAGGGGAAAAASGGGGAGGGPSGAP (SEQ ID NO: 74), GAPGGGGAGGGGAAAAASGGGGAGGGGAAAAASGGGGAGGGGAAAAASGGGP SGAP (SEQ ID NO: 76), and GAPGGGGAGGGGAAAAASGGGPSGGGGAAAAASGGGPSGGGGAAAAASGGGP S GAP (SEQ ID NO: 78).
2. The targeted therapeutic fusion protein of claim 1 , wherein the IGF-II peptide tag is an N-terminal tag or a C-terminal tag.
3. The targeted therapeutic fusion protein of claim 1 , wherein the IGF-II peptide tag comprises amino acids 8-67 of SEQ ID NO: 5.
4. The targeted therapeutic fusion protein of claim 1 , wherein the IGF-II peptide tag comprises a mutation at the position corresponding to position 37 of the polypeptide of SEQ ID NO: 5.
5. A composition comprising the targeted therapeutic fusion protein of claim 1 , and a pharmaceutically acceptable carrier, diluent or excipient.
6. A pharmaceutical composition suitable for treating a lysosomal storage disease comprising the targeted therapeutic fusion protein of claim 1 .
7. A method for treating Mucopolysaccharidosis Type IIB (Sanfilippo B syndrome) in a subject or reducing glycosaminoglycan levels in vivo comprising administering to the subject suffering from Sanfilippo B syndrome a therapeutically effective amount of a pharmaceutical composition comprising a fusion protein comprising a human α-N-acetylglucosaminidase (Naglu) protein comprising amino acids 1-743 or 24-743 of SEQ ID NO: 1, an IGF-II peptide tag having an amino acid sequence at least 70% identical to amino acids 8-67 of SEQ ID NO: 5 and a spacer peptide located between the Naglu protein and the IGF-II peptide tag, wherein the spacer peptide comprises the amino acid sequence selected from the group consisting of:
GAPGGGGSGGGGSGGGGSGGGGSGGGPSGGGGSGGGPSGAP (SEQ ID NO: 29), GAPGGGGSGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 31), GAPGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 33), GAPGGGGSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 35), GAPGGGGSGGGGSGGGGSGGGGSGGGPSGAP (SEQ ID NO: 37), GAPGGGGSGGGGSAAAASGGGGSGGGPSGAP (SEQ ID NO: 39), GAPGGGGSGGGGSAAAASGGGGSGGGGSAAAASGGGGSGGGGSAAAASGGGPSGAP (SEQ ID NO: 41), GAPGGGGSGGGGSAAAASGGGPSGGGGSAAAASGGGPSGGGGSAAAASGGGPSGAP (SEQ ID NO: 43), GAPGGGSPAPTPTPAPTPAPTPAGGGPSGAP (SEQ ID NO: 49), GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51), GAPGGGSPAEAAAKEAAAKEAAAKEAAAKEAAAKAPSGGGGAP (SEQ ID NO: 55), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGGGGAGGGPSGAP (SEQ ID NO: 64), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGGAGGGGAGGGPSGA P (SEQ ID NO:66), GAPGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGPSGAP (SEQ ID NO: 68), GAPGGGGAGGGGAGGGGAGGGPSGGGGAGGGGAGGGPSGGGGAGGGGAGGGPSGAP (SEQ ID NO: 70), GAPGGGGAGGGGAGGGGAGGGGAGGGPSGAP (SEQ ID NO: 72), GAPGGGGAGGGGAAAAASGGGGAGGGPSGAP (SEQ ID NO: 74), GAPGGGGAGGGGAAAAASGGGGAGGGGAAAAASGGGGAGGGGAAAAASGGGPSGA P (SEQ ID NO: 76), and GAPGGGGAGGGGAAAAASGGGPSGGGGAAAAASGGGPSGGGGAAAAASGGGPSGAP (SEQ ID NO: 78).
8. The method of claim 7 , wherein the IGF-II peptide tag is an N-terminal tag or a C-terminal tag.
9. The method of claim 7 , wherein the IGF-II peptide tag comprises amino acids 8-67 of SEQ ID NO: 5.
10. The method of claim 7 , wherein the IGF-II peptide tag comprises a mutation at the position corresponding to position 37 of the polypeptide of SEQ ID NO: 5.
11. The method of claim 7 , wherein the effective amount of the fusion protein is in the range of 2.5-20 mg per kilogram of body weight of the subject.
12. The method of claim 7 , wherein the fusion protein is administered intrathecally, optionally further comprising administering the fusion protein intravenously.
13. The method of claim 7 , wherein the fusion protein is administered intravenously.