Method for targeting lysosomal enzymes

• US 20090029467A1
• Filed: 05/30/2007
• Published: 01/29/2009
• Est. Priority Date: 04/30/2001
• Status: Active Grant

• Current Assignee: Biomarin Pharmaceutical Incorporated (Amgen Inc.)
• Original Assignee: ZyStor Therapeutics, Inc. (Amgen Inc.)
• Inventors: Beverley, Stephen M., LeBowitz, Jonathan H.
• Granted Patent: US 7,858,576 B2
• Time in Patent Office: Days
• US Class Current: 435/375
• 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

1-57. -57. (canceled)

58. A method of delivering a therapeutic enzyme to a lysosome in a glycosylation-independent manner, the method comprising:

(a) contacting a cell with the therapeutic enzyme associated with a glycosylation-independent lysosomal targeting (GILT) domain, wherein the GILT domain binds the cation-independent mannose-6-phosphate receptor on the surface of the cell in a mannose-6-phostphate-independent manner,

(b) delivering the therapeutic enzyme to the lysosome as a result of the binding between the GILT domain and the cation-independent mannose-6-phosphate receptor.

59. The method of claim 58, wherein the therapeutic enzyme is associated with the GILT domain in a fusion protein configuration.

60. The method of claim 58, wherein the GILT domain binds to the cation-independent mannose-6-phosphate receptor with a dissociation constant less than 10⁻

8 M at pH 7.4.

61. The method of claim 60, wherein the GILT domain binds to the cation-independent mannose-6-phosphate receptor with a dissociation constant between 10⁻

7 and 10¹¹ M at pH 7.4.

62. The method of claim 58, wherein the GILT domain comprises a mature human IGF-II.

63. The method of claim 58, wherein the GILT domain comprises a mutein of mature human IGF-II having an amino acid sequence at least 70% identical to mature human IGF-II and wherein the mutein binds to the human cation-independent mannose-6-phosphate receptor with a dissociation constant less than 10⁻

8 M at pH 7.4.

64. The method of claim 63, wherein the mutein comprises a deletion or a replacement of amino acids 1-7 of mature human IGF-II.

65. The method of claim 63, wherein the mutein is a fragment of mature human IGF-II.

66. The method of claim 58, wherein the cell is a mammalian cell.

67. The method of claim 66, wherein the mammalian cell is deficient in a lysosomal enzyme.

68. The method of claim 67, wherein the therapeutic enzyme is capable of rescuing the deficiency of the lysosomal enzyme.

69. The method of claim 66, wherein the mammalian cell is a human cell.

70. The method of claim 58, wherein the therapeutic enzyme is underglycosylated.