Delivery of biologically active polypeptides

• US 6,605,286 B2
• Filed: 04/16/1998
• Issued: 08/12/2003
• Est. Priority Date: 10/20/1995
• Status: Expired due to Term

• Current Assignee: Jeremy Mark Wells, Intrexon Actobiotics N.V. (Precigen, Inc.), Richard William Falla Le Page
• Original Assignee: Microbial Technics Limited, Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw
• Inventors: Steidler, Lothar, Remaut, Erik, Wells, Jeremy Mark, Le Page, Richard William Falla
• Primary Examiner(s): MINNIFIELD, NITA M
• Application Number:
  US09/060,878
• Publication Number:
  US 20010006642A1
• Time in Patent Office: 1,944 Days
• Field of Search: 424/184.1, 424/243.1, 424/192.1, 424/200.1, 424/258.1, 424/198.1, 424/199.1, 435/252.3, 435/320.1, 514/2, 514/3
• US Class Current: 424/243.1
• 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: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C07K 14/33 from Clostridium (G)
• CPC Class Codes: C07K 14/5412 IL-6
• CPC Class Codes: C07K 14/55 IL-2
• CPC Class Codes: C12N 15/74 Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
• CPC Class Codes: C12N 15/746 for lactic acid bacteria (Streptococcus; Lactococcus; Lactobacillus; Pediococcus; Enterococcus; Leuconostoc; Propionibacterium; Bifidobacterium; Sporolactobacillus)

Abstract

Biologically active polypeptides and/or antigens are delivered by administering to a subject a non-invasive or non-pathogenic bacterium which expresses one or more antigens or polypeptides. The non-invasive or non-pathogenic bacterium can be included in delivery systems or pharmaceutical formulations.

Claims

1. A method of delivering at least one heterologous biologically active polypeptide to a subject in need of same which comprises administering to the subject a non-invasive or non-pathogenic bacterium which constitutively expresses at least one heterologous biologically active polypeptide to illicit a biological response from the subject.

2. The method of claim 1 wherein the at least one heterologous biologically active polypeptide is an antigen.

3. The method of claim 2 wherein at least one antigen is heterologous to the bacterium.

4. The method of claim 3 wherein the heterologous polypeptide is derived from an eukaryote or its virus, or from a prokaryote or its virus.

5. The method of claim 1 wherein the bacterium is a Gram-positive bacterium.

6. The method of claim 5 wherein the Gram-positive bacterium is Listeria innocua, Staphylococcus xylosus, Staphylococcus carnosus, Streptococcus gordoni, a Lactococcus species or a Lactobacillus species.

7. The method of claim 6 wherein the Gram-positive bacterium is Lactococcus lactis.

8. The method of claim 5 wherein the bacterium is a non-pathogenic attenuated strain of a Gram-positive pathogenic bacterium.

9. The method of claim 8 wherein the bacterium is Listeria monocytogenes.

10. The method of claim 1 wherein the biologically active polypeptide is one which exerts endocrine activities affecting local or systemic metabolism.

11. The method of claim 1 wherein the biologically active polypeptide is one which regulates the activities of cells belonging to the immunohaemopoeitic system.

12. The method of claim 1 wherein the biologically active polypeptide is one which affects the viability, growth and differentiation of normal or neoplastic cells in the body or affecting the immune regulation or induction of acute phase inflammatory responses to injury and infection.

13. The method of claim 1 wherein the biologically active polypeptide is one which enhances or reduces resistance to infection of cells and tissues mediated by chemokines acting on their target cell receptors, or the proliferation of epithelial cells or the promotion of wound healing.

14. The method of claim 1 wherein the biologically active polypeptide is insulin, growth hormone, prolactin, calcitonin, luteinising hormone, parathyroid hormone, somatostatin, thyroid stimulating hormone or vasoactive intestinal polypeptide.

15. The method of claim 1 wherein the biologically active polypeptide is a structural group 1 cytokine adopting an antiparallel 4α

helical bundle structure.

16. The method of claim 1 wherein the biologically active polypeptide is a structural group 2 cytokine which forms symmetric homotrimers and the subunits take up the conformation of β

-jelly roll described for the TNF family of cytokines, the IL-1 family of cytokines, the fibroblast growth factor family, the platelet derived growth factors, transforming growth factor β

or nerve growth factors.

17. The method of claim 1 wherein the biologically active polypeptide is a structural group 3 cytokine comprising short chain α

/β

molecules, which are produced as transmembrane pre-cursor molecules which each contain at least one EGF domain in the extracellular region.

18. The method of claim 1 wherein the biologically active polypeptide is a structural group 4 cytokine which exhibits mosaic structures.

19. The method of claim 1 wherein the biologically active polypeptide is a receptor or antagonist for a biologically active polypeptide.

20. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which regulates the survival, growth, differentiation, effector functions or susceptibility to infection of cells or tissues.

21. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which boosts an immune response against tumour cells or an infection colonising a mucosal surface or adjacent or distant tissue.

22. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which modulates the type of immune response (antibody versus cell-medicated) against a pathogenic infectious agent.

23. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which modulates the infiltration of normal tissues with inflammatory or tumour cells.

24. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which controls the rate of growth, rate of invasion or survival of tumour cells.

25. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which induces apoptosis in tumour cells.

26. The method of claim 1 wherein the bacterium expresses an antigen or polypeptide which downregulates an immune response.

27. The method of claim 1, wherein the bacterium expresses an antigen or polypeptide which treats an immune dysregulative diseases state.

28. The method of claim 15, wherein the structural group I cytokine is IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-11, IL-12, IL-13, GM-CSF, M-CSF, SCR, IFN-γ

, EPO, G-CSF, LIF, OSM, CNTF, GH, PRL or IFNα

/β

.

29. The method of claim 28, wherein the structural group I cytokine is IL-2 or IL-6.

30. The method of claim 27, wherein the immune dysregulative diseases is an allergic autoimmune disease state.