Site-specific antibody-mediated activation of proapoptotic cytokines: AMAIZEe (antibody-mediated apoptosis inducing cytokines)

• US 20040033511A1
• Filed: 03/14/2003
• Published: 02/19/2004
• Est. Priority Date: 09/15/2000
• Status: Active Grant

• Current Assignee: Biontech AG
• Original Assignee: Biontech AG
• Inventors: Wuest, Thomas, Pfizenmaier, Klaus, Wajant, Harald, Moosmayer, Dieter
• Granted Patent: US 8,980,265 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61K 47/6899 Antibody-Directed Enzyme Prodrug Therapy [ADEPT]
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 14/525 Tumour necrosis factor [TNF]
• CPC Class Codes: C07K 14/70575 NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 NGF C07K14/48, TNF C07K14/525
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Subject matter of the invention are antibody-cytokine fusion proteins having proapoptotic and immune modulating properties, but wherein the cytokine moiety a priori has a bioactivity which is very low or restricted to certain receptor subtypes. These reagent exert their full biological activity via the corresponding cytokine receptor(s) only after antibody-mediated binding of the fusion protein to a specific cell membrane-expressed target molecule. By suitable choice of the antibody specificity, the cytokine activity is directed to the tissue, e.g. tumour tissue, to be treated, and a therapeutic agent can be produced being specifically designed/optimised for the respective indication/tumour entity.

Claims

1. A polypeptide comprising:

(i) a segment (1) comprising biological activity for a specific target molecule;

(ii) N-terminal of segment (1) a segment (2) comprising a peptide linker; and

(iii) a segment (3) comprising an antibody or a fragment thereof that selectively recognizes a specific target molecule on a cell surface,

wherein the polypeptide has no or limited biological activity without site-specific or selective binding of segment (3) to the specific target molecule on the cell surface.

2. The polypeptide according to claim 1, wherein segment (1) comprises an amino acid sequence of a cytokine or a fragment thereof.

3. The polypeptide according to claim 2, wherein the cytokine comprises an extracellular domain, a functional variant of an extracellular domain, or a functional fragment of an extracellular domain of a member of the TNF family.

4. The polypeptide according to claim 2, wherein the cytokine comprises an extracellular domain, a functional variant of an extracellular domain or a functional fragment of an extracellular domain of TRAIL (TNF Related Apoptosis Inducing Ligand, AA 95-281, NCBI Accession No. AAC50332) or of FasL (AA 139-281, NCBI Accession No. AAC50124).

5. The polypeptide according to claim 1, wherein the polypeptide recognises a cell membrane-bound cytokine receptor as the specific target molecule.

6. The polypeptide according to claim 1, wherein segment (2) links segments (3) and (1), and wherein segment (2) is an intrinsically defined polymerisation module.

7. The polypeptide according claim 6, wherein the polymerisation module comprises a naturally occurring or synthetically produced peptide comprising dimerisation properties, trimerisation properties, hexamerisation properties or a combination thereof.

8. The polypeptide according to claim 1, wherein fragment (3) comprises an antigen binding antibody or an antigen binding antibody fragment.

9. The polypeptide according to claim 1, wherein segment (3) comprises an antibody or an antibody fragment of a mammal, or a humanised antibody or a humanised antibody fragment.

10. The polypeptide according to claim 9, wherein the antibody fragment comprises a scFv, Fab or complete immunoglobulin.

11. A nucleic acid construct comprising a nucleotide sequence encoding a polypeptide of claim 1.

12. A vector comprising the nucleic acid construct of claim 11.

13. A host cell comprising the nucleic acid of claim 11, the vector of claim 12, or a combination thereof.

14. A method for the isolation of a polypeptide according to claim 1 comprising:

(a) producing a vector according to claim 12 or a nucleic acid construct according to claim 11;

(b) transfecting cells with the vector or nucleic acid construct obtained according to step (a);

(c) culturing the cells transfected according to step (b); and

(d) isolating polypeptides expressed under appropriate conditions from the cultured cells according to step (c), culture supernatant, or a combination thereof.

15. A method for treating cancer, infectious diseases, metabolic diseases, inflammatory conditions, or autoimmune diseases comprising administration of a polypeptide of claim 1, a nucleic acid construct of claim 11, a vector of claim 12, or a host cell of claim 13.

16. A composition comprising a polypeptide according to claim 1, a nucleic acid construct according to claim 11, a vector according to claim 12, a host cell according to claim 13, or a combination thereof, and a pharmaceutically acceptable excipient, additive, and/or carrier.

17. A method for treating cancer, infectious diseases, metabolic diseases, inflammatory conditions, or autoimmune diseases comprising administration of the composition of claim 16.

18. The polypeptide according to claim 7, wherein the polymerization module comprises an immunoglobulin hinge region and the CH3 domain of a human immunoglobulin gene (AA 363-489, human IgG1, NCBI Accession No. AAF21613), a domain of a Tenascin molecule with trimerization properties, or an extended domain of a Tenascin molecule with hexamerisation properties (AA 34-139, Swiss Prot. Accession No. P10039, chicken, Swiss Prot. Accession No. P24821, human).

19. The polypeptide according to claim 9, wherein the mammal is murine or human.

20. The method according to claim 15, wherein the cancer comprises a solid or lymphatic tumour.

21. The method according to claim 15, wherein the autoimmune disease comprises a rheumatoid arthritic disease.

22. The method according to claim 17, wherein the cancer comprises a solid or lymphatic tumour.

23. The method according to claim 17, wherein the autoimmune disease comprises a rheumatoid arthritic disease.