Anti-human tenascin monoclonal antibody

• US 20050106145A1
• Filed: 02/20/2003
• Published: 05/19/2005
• Est. Priority Date: 02/26/2002
• Status: Active Grant

• Current Assignee: Alfasigma S.p.A.
• Original Assignee: Sigma-Tau Industrie Farmaceutiche Riunite SPA (Alfasigma S.p.A.)
• Inventors: Anna Maria, Anastasi, Rita, De Santis
• Granted Patent: US 7,438,908 B2
• Time in Patent Office: Days
• US Class Current: 424/145.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: G01N 2333/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/975 Kit

Abstract

A novel anti-human tenascin ST2146 monoclonal antibody is described endowed with high affinity with the native antigen and high tumor selectivity. The cST2146 hybridoma is stably producing the antibody in high density culture conditions and is suitable for the industrial development of ST2146-based products. ST2146 exhibits properties exploitable for both therapeutic and diagnostic applications.

Claims

1. Monoclonal antibody anti-human tenascin whose light and heavy chain variable region sequences are SEQ ID NO:

1 and SEQ ID NO;

2, respectively, and the proteolytic fragments thereof binding to an antigenic epitope within the EGF-like repeat of human tenascin C.

2. The antibody and the fragments thereof according to claim 1 further containing additional markers and/or diagnostic agents.

3. Recombinant derivatives of the antibody and the fragments thereof of claim 1.

4. Recombinant derivatives according to claim 3, wherein the murine constant region is replaced by the human counterpart or a biologically or pharmacologically active moiety.

5. Recombinant derivatives according to claim 4, wherein said biologically active moiety is selected from the group consisting of:

a member of the avidin family, a growth factor useful for stimulating tumor-directed immunological effectors.

6. Recombinant derivatives according to claim 4, wherein said pharmacologically active moiety is selected from the group consisting of:

A toxin, a superantigen, a cytokine or any other protein useful in enhancing the antitumor therapeutical efficacy.

7. Conjugate derivatives of the antibody and the fragments thereof of claim 1.

8. Conjugate according to claim 7, wherein a biologically or pharmacologically active moiety is linked to said antibody or the fragments thereof.

9. Conjugate according to claim 8, wherein said biologically active moiety is selected from the group consisting of a growth factor useful for stimulating tumor-directed immunological effectors or said pharmacologically active moiety is selected from the group consisting of a toxin, a superantigen, a cytokine or any other protein useful in enhancing the antitumor therapeutical efficacy, antitumor drugs, radioisotopes.

11. A biotinylated derivative of the antibody of claim 1, or fragments or recombinant derivatives or conjugate thereof or immunoglobulin thereof.

12. The hybridoma cell line producing the antibody of claim 1, deposited with Advanced Biotechnology Center on 29 Jan. 2002 under the provision of Budapest Treaty, with the number PD02003.

13. Process for the preparation of antibody, comprising cultivating the hybridoma of claim 12 and isolating said antibody.

14. A method of detecting of disease comprising contacting a patient sample with an antibody of claim 1 or a fragment or recombinant derivative or conjugate thereof and detecting binding of the same to a species of said sample as an indication of said disease.

15. The method of claim 14, wherein said disease is a tumor.

16. The method of claim 15, wherein said tumor is selected from the group consisting of gliomas, mammary, lung carcinomas, fibrosarcomas and squamous cell carcinomas.

17. The method of claim 14, comprising an in-vivo imaging technique.

22. Therapeutic kit composed of 5 vials, whose first vial contains the the biotinylated derivative of claim 11, the second vial contains an avidin, the third vial contains biotinylated albumine, the fourth vial contains a streptavidin, the fifth vial contains radiolabelled biotin or biotin derivative.

23. Therapeutic kit according to claim 22, wherein said vials are suitable for human injection.

24. encoding the antibody of claim 1 or fragment thereof or recombinant derivative or immunoglobulin thereof.

26. Vector containing DNA of claim 24.

27. Host cells containing vectors of claim 26.

28. A method of determining dievels of circulating tenascin comprising contacting an antibody of claim 1 or a fragment or recombinant derivative or conjugate thereof with a second tenascin-specific antibody in a sandwich assay in the presence of a patient sample and determing of the level of circulating tenascin.

29. Diagnostic kit comprising the antibody of claim 1 or fragments, or recombinant derivatives or conjugates thereof or an immunoglobulin thereof.

30. A container, optionally containing multiple compartments, comprising the biotinylated antibody or fragments thereof or immunoglobulin of claim 11, buffers and reagents suitable for use in a therapeutic kit for a three-step pre-targeting method.

31. The container according to claim 30, further comprising a separate tenascin-specific antibody preferentially directed towards the A-D fragment.

32. The container according to claim 30, further comprising a separate tumor specific antibody.

33. The antibody of claim 1 or in combination with a second tenascin-specific antibody in a sandwich ELISA in vitro assay under conditions where said second antibody binds to a second antigenic epitope of tenascin, said in vitro ELISA assay useful for determining the level of circulating tenascin.

34. Pharmaceutical composition comprising the antibody of claim 1 and/or a fragment and/or a recombinant derivative and/or a conjugate thereof and/or an immunoglobulin thereof, in admixture with at least one pharmaceutically acceptable excipient and/or vehicle.

10. Immunoglobulin molecule which specifically binds tenascin.

18-21. -21. (canceled)

25. Specific CDRs and proteins comprising or containing CDRs.