Fusion proteins and polynucleotides encoding gelonin sequences

• US 20030166196A1
• Filed: 04/23/2002
• Published: 09/04/2003
• Est. Priority Date: 11/04/1991
• Status: Abandoned Application

• Current Assignee: Research & Development Foundation
• Original Assignee: Xoma Technology Ltd. (Xoma Corp.)
• Inventors: Carroll, Stephen F., Better, Marc D.
• Application Number:
  US10/127,890
• Publication Number:
  US 20030166196A1
• Time in Patent Office: Days
• US Class Current: 435/188.5
• 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 47/6825 Ribosomal inhibitory proteins, i.e. RIP-I or RIP-II, e.g. Pap, gelonin or dianthin
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/16 against material from plants
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention provides purified and isolated polynucleotides encoding Type I ribosome-inactivating proteins (RIPS) and analogs of the RIPs having a cysteine available for disulfide bonding to targeting molecules. Vectors comprising the polynucleotides and host cells transformed with the vectors are also provided. The RIPs and RIP analogs are particularly suited for use as components of cytotoxic therapeutic agents of the invention which include gene fusion products and immunoconjugates. Cytotoxic therapeutic agents or immunotoxins according to the present invention may be used to selectively eliminate any cell type to which the RIP component is targeted by the specific binding capacity of the second component of the agent, and are suited for treatment of diseases where the elimination of a particular cell type is a goal, such as autoimmune disease, cancer and graft-versus-host disease.

Claims

1. A fusion protein comprising (a) a gelonin amino acid sequence that has enzymatic activity and (b) a targeting sequence that allows the internalization of said fusion protein, wherein said targeting sequence is an antibody, an antigen-binding portion of an antibody, a hormone, a lymphokine or a growth factor.

2. The fusion protein of claim 1, wherein said gelonin sequence is that of SEQ Id no. 2.

3. The fusion protein of claim 1, wherein said gelonin sequence is that of SEQ ID NO. 101.

4. The fusion protein of claim 1, further comprising a linker sequence between said gelonin sequence and said targeting sequence, wherein said gelonin possesses enzymatic activity, said antibody is capable of recognizing antigen and said hormone, lymphokine or growth factor is capable of binding to a cell that has a receptor for said hormone lymphokine or growth factor.

5. The fusion protein of claim 4, wherein said linker sequence is that of SEQ ID NO. 56 or SEQ ID NO. 57.

6. The fusion protein of any one of claim 1-5, wherein said targeting sequence is an antibody.

7. The fusion protein of any one of claims 1-5, wherein said targeting sequence is an antigen-binding portion of an antibody.

8. The fusion protein of claim 7, wherein said antigen-binding portion of said antibody is an Fab.

9. The fusion protein of claim 7, wherein, wherein said antigen-binding portion of said antibody is an Fab′

.

10. The fusion protein of claim 7, wherein said antigen-binding portion of said antibody is an F(ab′

)₂.

11. The fusion protein of claim 7, wherein said antigen-binding portion of said antibody is an Fv.

12. The fusion protein of claim 7, wherein said antigen-binding portion of said antibody has a single variable domain.

13. The fusion protein of claim 7, wherein said antibody is a single-chain antibody.

14. The fusion protein of claim 7, wherein said fusion protein is multivalent.

15. The fusion protein of any one of claims 1-5, wherein said targeting sequence is a hormone.

16. The fusion protein of any one of claims 1-5, wherein said targeting sequence is a lymphokine.

17. The fusion protein of any one of claims 1-5, wherein said targeting sequence is a growth factor.