Targeted therapeutic lysosomal enzyme fusion proteins and uses thereof

Patent metadata

Field	Value
Publication number	US-9376480-B2
Application number	US-201314092336-A
Country	US
Kind code	B2
Filing date	Nov 27, 2013
Priority date	Nov 27, 2012
Publication date	Jun 28, 2016
Grant date	Jun 28, 2016

Abstract

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.

First claim

What is claimed:

1. A targeted therapeutic fusion protein comprising (i) a human α-N-acetylglucosaminidase (Naglu) protein comprising amino acids 1-743 or 24-743 of SEQ ID NO:1, (ii) a peptide tag comprising SEQ ID NO: 2 or amino acids 8-67 of SEQ ID NO:5and (iii) a spacer peptide located between the Naglu protein and the peptide tag, wherein the spacer peptide comprises the amino acid sequence GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51).

2. The targeted therapeutic fusion protein of claim 1 , wherein the peptide tag is an N-terminal tag or a C-terminal tag.

3. The targeted therapeutic fusion protein of claim 1 , wherein the peptide tag comprises amino acids 8-67 of SEQ ID NO:5.

4. The targeted therapeutic fusion protein of claim 1 , wherein the peptide tag comprises SEQ ID NO:2.

5. A pharmaceutical composition suitable for treating a lysosomal storage disease comprising the targeted therapeutic fusion protein of claim 1 and a pharmaceutically acceptable carrier, diluent or excipient.

6. The targeted therapeutic fusion protein of claim 1 , wherein the peptide tag consists of amino acids 8-67 of SEQ ID NO:5.

7. The targeted therapeutic fusion protein of claim 1 , wherein the spacer peptide consists of the amino acid sequence of SEQ ID NO:51.

8. A targeted therapeutic fusion protein comprising (i) a human α-N-acetylglucosaminidase (Naglu) protein consisting of amino acids 1-743 or 24-743 of SEQ ID NO:1, (ii) a peptide tag consisting of SEQ ID NO:2 or amino acids 8-67 of SEQ ID NO:5 and (iii) a spacer peptide located between the Naglu protein and the peptide tag, wherein the spacer peptide consists of the amino acid sequence GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51).

9. The targeted therapeutic fusion protein of claim 8 that consists of (i) a human α-N-acetylglucosaminidase (Naglu) protein consisting of amino acids 24-743 of SEQ ID NO:1, (ii) a peptide tag consisting of amino acids 8-67 of SEQ ID NO:5 and (iii) a spacer peptide located between the Naglu protein and the peptide tag, wherein the spacer peptide consists of the amino acid sequence GAPGGGSPAPAPTPAPAPTPAPAGGGPS GAP (SEQ ID NO: 51).

10. The targeted therapeutic fusion protein of claim 8 that consists of (i) a human α-N-acetylglucosaminidase (Naglu) protein consisting of amino acids 24-743 of SEQ ID NO:1, (ii) a peptide tag consisting of SEQ ID NO:2 and (iii) a spacer peptide located between the Naglu protein and the peptide tag, wherein the spacer peptide consists of the amino acid sequence GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51).

11. A pharmaceutical composition suitable for treating a lysosomal storage disease comprising the targeted therapeutic fusion protein of claim 8 and a pharmaceutically acceptable carrier, diluent or excipient.

12. A pharmaceutical composition suitable for treating a lysosomal storage disease comprising the targeted therapeutic fusion protein of claim 9 and a pharmaceutically acceptable carrier, diluent or excipient.

13. A pharmaceutical composition suitable for treating a lysosomal storage disease comprising the targeted therapeutic fusion protein of claim 10 and a pharmaceutically acceptable carrier, diluent or excipient.

14. A method for treating Mucopolysaccharidosis Type IIIB (Sanfilippo B Syndrome) in a subject or reducing glycosaminoglycan levels in vivo comprising administering to the subject suffering from Sanfilippo B Syndrome a therapeutically effect amount of a pharmaceutical composition comprising a targeted therapeutic fusion protein comprising (i) a human α-N-acetylglucosaminidase (Naglu) protein comprising amino acids 1-743 or 24-743of SEQ ID NO: 1, (ii) a peptide tag comprising SEQ ID NO: 2 or amino acids 8-67 of SEQ ID NO: 5, and (iii) a spacer peptide located between the Naglu protein and the peptide tag, wherein the spacer peptide comprises the amino acid sequence GAPGGGSPAPAPTPAPAPTPAPAGGGPSGAP (SEQ ID NO: 51).

15. The method of claim 14 , wherein the peptide tag is an N-terminal tag or a C-terminal tag.

16. The method of claim 14 , wherein the peptide tag comprises amino acids 8-67 of SEQ ID NO: 5.

17. The method of claim 14 , wherein the peptide tag comprises SEQ ID NO: 2.

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

19. The method of claim 14 , wherein the pharmaceutical composition is administered intrathecally and/or intravenously.

20. The method of claim 19 , wherein the administering comprises introducing the pharmaceutical composition into a cerebral ventricle, lumbar area, or cisterna magna.

21. The method of claim 14 , wherein the pharmaceutical composition is administered intravenously.

Assignees

• Biomarin Pharm Inc

Inventors

• Aoyagi-Scharber Mika
• Christianson Teresa Margaret
• Dvorak-Ewell Melita
• Wendt Daniel J
• Long Shinong
• Lebowitz Jonathan
• Gold Daniel Solomon

Classifications

• C12N9/2402 Primary

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• A61K38/00 Primary

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C07K2319/06

  containing a lysosomal/endosomal localisation signal · CPC title

• C12Y302/0105

  Alpha-N-acetylglucosaminidase (3.2.1.50) · CPC title