Method for producing proteins suitable for treating lysosomal storage disorders

• US 7,241,442 B2
• Filed: 12/11/2003
• Issued: 07/10/2007
• Est. Priority Date: 09/28/2001
• Status: Expired due to Term

• Current Assignee: St Jude Children's Research Hospital Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: St Jude Children's Research Hospital Incorporated (Abbott Laboratories Incorporated)
• Inventors: Bonten, Erik Jacobus, D'Azzo, Alessandra
• Primary Examiner(s): Saidha; Tekchand
• Assistant Examiner(s): Fronda; Christian L.
• Application Number:
  US10/733,501
• Publication Number:
  US 20040126370A1
• Time in Patent Office: 1,307 Days
• Field of Search: 435/69.1, 435/183, 435/195, 435/200, 435/201, 424/94.1, 424/94.6, 424/94.61
• US Class Current: 424/94.1
• CPC Class Codes: A61K 38/00 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 Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12N 9/48 acting on peptide bonds (3.4)
• CPC Class Codes: C12Y 302/01018 Exo-alpha-sialidase (3.2.1.18), i.e. trans-sialidase
• CPC Class Codes: C12Y 304/16005 Carboxypeptidase C (3.4.16.5), i.e. carboxypeptidase Y

Abstract

The present invention is based on the discovery that proteins produced in insect cell cultures are glycosylated in a unique manner that causes them to be selectively imported by cells that express mannose receptors on their membranes, particularly macrophages. Proteins that are selectively imported into cells containing mannose receptors are provided, as well as pharmaceutical compositions containing such proteins and methods for producing such proteins. Application of the present invention to produce proteins useful for treating lysosomal storage disorders is also disclosed. Engineering of cells to express mannose receptors so that they will selectively import proteins produced in insect cells is also taught, as well as a protein targeting system using such cells and proteins. Finally, an improved elution buffer for the purification of proteins produced in insect cells from a Concanavalin A column is provided.

Claims

1. A method of treating a subject suffering from a lysosomal storage disorder other than Fabry Disease caused by a deficiency of a specific protein comprising:

(a) producing said protein in an insect cell culture such that said protein will be selectively imported into macrophages when administered to said subject, and

(b) administering a therapeutically effective amount of said protein to said subject.

2. The method of claim 1 wherein said protein is produced in an insect cell culture using a baculovirus expression system.

3. The method of claim 1 wherein said insect cell culture is derived from the species Spodoptera frugiperda.

4. The method of claim 3 wherein said insect cell culture is an Sf9 cell culture.

5. The method of claim 1 wherein said lysosomal storage disorder and associated protein useful for treating said lysosomal storage disorder are selected from the group consisting of Pompe Disease and acid α

-1,4 glucosidase, Pompe Disease and acid α

-1,6 glucosidase, GM1 gangliosidosis and β

-galactosidase, Tay-Sachs disease and β

-hexasaminidase A, GM2 gangliosialidosis;

AB Variant and GM₂, Activator Protein, Sandhoff Disease and β

-hexosaminidase A, Sandhoff Disease and β

-hexosaminidase B, Gaucher Disease and glucocerebrosidase, Gaucher Disease and β

-galacosidase, Krabbe Disease and galactosylcerebrosidase, Niemann-Pick Type A and acid sphingomyelinase, Niemann-Pick Type B and acid sphingomyelinase, Farber Disease and acid ceramidase, Wolman Disease and acid lipase, Cholesterol Ester Storage Disease and acid lipase, Hurler Syndrome and α

-L-iduronidase, Scheie Syndrome and α

-L-iduronidase, Hurler-Scheie and α

-L-iduronidase, Hunter Syndrome and iduronate 2-sulfatase, Sanfilippo A and α

-N-acetylglucosaminidase, Sanfilippo B and α

-N-acetylglucosaminidase, Sanfilippo C and acetyl-CoA-glucosaminide acetyltransferase, Sanfilippo D and N -acetylglucosamine-6-sulfatase, Morquio A and N-acetylglucosamine-6-sulfate sulfatase, Morquio B and β

-galactosidase, Maroteaux-Lamy and arysuylfatase B, Sly Syndrome and β

-glucuronidase, Metachromatic Leukodystrophy and arylsulfatase A, Multiple Sulfatase Deficiency and arylsulfatase A, Multiple Sulfatase Deficiency and arylsulfatase B, Multiple Sulfatase Deficiency and arylsulfatase C, Sialidosis and α

-Neuraminidase, I -cell Disease and UDP GlcNAc;

lysosomal-enzyme N-acetyglucosamine-1-phosphotransferase, Pseudo-Hurler Polydistrophy and UDP GlcNAc;

lysosomal-enzyme N-acetylglucosamine-1-phosphotransferase, Mucolipidosis IV and mucolipin-1, α

-Mannosidosis and α

-mannosidase, β

-Mannosidosis and β

-mannosidase, Fucosidosis and α

-L-fucosidase, Aspartylglucosaminuria and N-aspartyl-β

-glucosaminidase, Galactosialidosis and protective protein/cathepsin A, Galactosialidosis and neuraminidase, Galactosialidosis and β

-galactosidase, Schindler Disease and α

-N-acetyl -galactosaminidase, Cystinosis and cystine transport protein, Salla Disease and sialin, Infantile Sialic Acid Storage Disorder and sialin, Infantile Neuronal Ceroid Lipofuscinosis and palmitoly-protein thioesterase, Prosaposin and Saposin A, Prosaposin and Saposin B, Prosaposin and Saposin C, and Prosaposin and Saposin D.

6. The method of claim 5 wherein said lysosomal storage disorder is Galactosialidosis and said protein useful for treating said lysosomal storage disorder is protective protein/cathepsin A.

7. The method of claim 5 wherein said lysosomal storage disorder is Sialidosis and said protein useful for treating said lysosomal storage disorder is α

-Neuraminidase.