Method for targeting lysosomal enzymes

• US 7,858,576 B2
• Filed: 05/30/2007
• Issued: 12/28/2010
• Est. Priority Date: 04/30/2001
• Status: Expired due to Term

• Current Assignee: Biomarin Pharmaceutical Incorporated (Amgen Inc.)
• Original Assignee: ZyStor Therapeutics, Inc. (Amgen Inc.)
• Inventors: Beverley, Stephen M., LeBowitz, Jonathan H.
• Primary Examiner(s): Nickol; Gary B.
• Assistant Examiner(s): Gamett; Daniel C
• Application Number:
  US11/807,729
• Publication Number:
  US 20090029467A1
• Time in Patent Office: 1,308 Days
• Field of Search: None
• US Class Current: 514/1.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/515 Animal cells
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/54 Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
• CPC Class Codes: A61K 38/30 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2 insulin-like growth factor binding protein A61K38/1754
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 47/551 one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/642 the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate
• CPC Class Codes: A61K 47/6425 the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: C07K 14/65 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• CPC Class Codes: C07K 2319/02 : containing a signal sequence
• CPC Class Codes: C07K 2319/06 : containing a lysosomal/endosomal localisation signal
• CPC Class Codes: C07K 2319/21 : containing a His-tag
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/50 : containing protease site
• CPC Class Codes: C07K 2319/61 : containing an enzyme fusion for detection (lacZ, luciferase)
• CPC Class Codes: C07K 2319/74 : containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C07K 2319/75 : containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• CPC Class Codes: C12N 15/62 : DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 9/0051 : acting on a sulfur group of donors (1.8)
• CPC Class Codes: C12N 9/2402 : hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01022 : Alpha-galactosidase (3.2.1.22)

Abstract

Targeted therapeutics that localize to a specific subcellular compartment such as the lysosome are provided. The targeted therapeutics include a therapeutic agent and a targeting moiety that binds a receptor on an exterior surface of the cell, permitting proper subcellular localization of the targeted therapeutic upon internalization of the receptor. Nucleic acids, cells, and methods relating to the practice of the invention are also provided.

Claims

1. A method of targeting a therapeutic enzyme to a lysosome, the method comprising:

(a) providing a targeted therapeutic comprising a therapeutic enzyme and a lysosomal-targeting domain that binds the cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner;

wherein the lysosomal-targeting domain comprises a mature human IGF-II;

(b) contacting the targeted therapeutic with a cell deficient in a lysosomal enzyme, wherein the cell expresses the cation-independent mannose-6-phosphate receptor on the surface, thereby targeting the therapeutic enzyme to the lysosome as a result of the binding between the lysosomal-targeting domain and the cation-independent mannose-6-phosphate receptor.

2. The method of claim 1, wherein the therapeutic enzyme is a lysosomal enzyme.

3. The method of claim 2, wherein the lysosomal enzyme is human acid-α

1,4-glucosidase.

4. A method of targeting a therapeutic enzyme to a lysosome, the method comprising:

a) providing a targeted therapeutic comprising a therapeutic enzyme and a lysosomal-targeting domain that binds the cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner;

wherein the lysosomal-targeting domain comprises a mutein of mature human IGF-II having an amino acid sequence at least 70% identical to mature human IGF-II;

b) contacting the targeted therapeutic with a cell deficient in a lysosomal enzyme, wherein the cell expresses the cation-independent mannose-6-phosphate receptor on the surface,

thereby targeting the therapeutic enzyme to the lysosome as a result of the binding between the lysosomal-targeting domain and the cation-independent mannose-6-phosphate receptor.

5. The method of claim 4, wherein the mutein comprises a deletion or a replacement of amino acids 1-7 of mature human IGF-II.

6. The method of claim 5, wherein the mutein comprises a replacement of amino acids 1-7 of mature human IGF-II with one or more amino acid residues.

7. The method of claim 4, wherein the mutein is a fragment of mature human IGF-II.

8. The method of claim 4, wherein the therapeutic enzyme is a lysosomal enzyme.

9. The method of claim 8, wherein the lysosomal enzyme is human acid-α

1,4-glucosidase.

10. A method of treating a lysosomal storage disease comprising administering to a subject in need of treatment a targeted therapeutic comprising a lysosomal enzyme and a lysosomal-targeting domain that binds the cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner, wherein the lysosomal-targeting domain comprises a mature human IGF-II.

11. The method of claim 10, wherein the lysosomal enzyme is human acid-α

1,4-glucosidase.

12. A method of treating a lysosomal storage disease comprising administering to a subject in need of treatment a targeted therapeutic comprising a lysosomal enzyme and a lysosomal-targeting domain that binds the cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner, wherein the lysosomal-targeting domain comprises a mutein of mature human IGF-II having an amino acid sequence at least 70% identical to mature human IGF-II.

13. The method of claim 12, wherein the mutein comprises a deletion or a replacement of amino acids 1-7 of mature human IGF-II.

14. The method of claim 13, wherein the mutein comprises a replacement of amino acids 1-7 of mature human IGF-II with one or more amino acid residues.

15. The method of claim 12, wherein the lysosomal enzyme is human acid-α

1,4-glucosidase.