Transgenically produced Antithrombin III

• US 6,441,145 B1
• Filed: 08/28/1998
• Issued: 08/27/2002
• Est. Priority Date: 02/21/1995
• Status: Expired due to Term

• Current Assignee: rEVO Biologics, Inc. (Agence des Participations de l’Etat)
• Original Assignee: Genzyme Transgenics Corporation
• Inventors: DiTullio, Paul, Cole, Edward S., Meade, Harry
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US09/143,155
• Time in Patent Office: 1,460 Days
• Field of Search: 800/7, 800/21, 800/14, 800/18, 536/24.5, 514/2-21, 435/69.1, 530/380, 530/381, 530/386, 530/392, 530/393, 530/395, 530/832, 930/240
• US Class Current: 530/393
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• 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: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: C07K 14/8128 Antithrombin III
• CPC Class Codes: Y10S 530/832 Milk; colostrum

Abstract

This invention relates to transgenically produced human Antithrombin III (tgATIII). The human ATIII produced by the transgenic process of the present invention has a monosaccharide composition which comprises N-acetylgalactosamine (GaINAc) along with fucose, N-acetylglucosamine, galactose, mannose, and N-acetylneuraminic acid/N-glycolyneuraminic acid. The monosaccharide composition differs with that of plasma derived ATIII (phATIII). It has been found that tgATIII has an increased clearance rate when compared to phATIII.

Claims

1. Mammary gland produced antithrombin III having a monosaccharide composition which comprises GalNAc.

4. The mammary gland produced antithrombin III of claim 1 further comprising fucose.

5. The mammary gland produced antithrombin III of claims 1, 2, or 4 wherein the antithrombin III further comprises a higher level of mannose than plasma derived antithrombin III.

6. The mammary gland produced antithrombin III of claim 1, 2, 3, or 4 wherein the antithrombin III is produced in the mammary glands of a transiemc goat.

2. Mammary gland produced antithrombin III having a monosaccharide composition which comprises fucose.

3. Mammary gland produced antithrombin III having a monosaccharide composition which comprises a higher level of mannose than plasma derived antithrombin III.

7. A method for producing antithrombin III in mammalian milk, comprising:

a. providing a transgenic mammal that expresses in its mammary tissue a transgene which encodes a human antithrombin III with a monosaccharide composition which comprises GalNAc, wherein said human antithrombin III is secreted into the milk of the mammal; and

b. collecting milk from the transgenic animal which contains the human antithrombin III;

c. to thereby obtain human antithrombin III with a monosaccharide composition which includes GalNAc.

8. The method of claim 7, further comprising isolating human ATIII from the milk.

9. The method of claim 7, wherein the transgenic mammal is goat.

10. The method of claim 7, wherein the transgenic mammal is a mouse.

11. A glycosylated human antithrombin III which is produced in mammary gland of a non-human transgenic mammal, wherein the antithrombin III comprises a monosaccharide composition which comprises GalNAc.

12. A glycosylated human antithrombin III which is produced in the mammary gland of a non-human transgenic mammal, wherein the antithrombin III comprises a monosaccharide composition which comprises fucose.

13. A glycosylated human antithrombin III which is produced in the mammary gland of a non-human transgenic mammal, wherein the antithrombin III comprises a monosaccharide composition which comprises a higher level of mannose than plasma derived antithrombin III.

14. The glycosylated human antithrombin III of claim 13 wherein the antithrombin III has a higher affinity for heparin binding as compared to plasma derived antithrombin III.

15. A method for producing antithrombin III in mammalian milk, comprising:

providing a transgenic mammal that expresses in its mammary tissue a transgene which encodes a human antithrombin III with a monosaccharide composition which comprises fucose, wherein said human antithrombin III is secreted into the milk of the mammal; and

collecting milk from the transgenic animal which contains the human antithrombin III

to thereby obtain human antithrombin III with a monosaccharide composition which includes fucose.

16. The method of claim 15, wherein the transgenic mammal is a goat.

17. The method of claim 15, wherein the transgenic mammal is a mouse.

18. A method for producing antithrombin III in mammalian milk, comprising:

providing a transgenic mammal that expresses in its mammary tissue a transgene which encodes a human antithrombin III having a monosaccharide composition which comprises a higher level of mannose than plasma derived antithrombin III, wherein said human antithrombin III is secreted into the milk,of the mammal; and

collecting milk from the transgenic animal which contains the human antithrombin III

to thereby obtain human antithrombin III having a monosaccharide composition which includes a higher level of mannose than plasma derived antithrombin III.

19. The method of claim 18, wherein the transgenic mammal is a goat.

20. The method of claim 18, wherein the transgenic mammal is a mouse.