Chimeric proteins and uses thereof

• US 8,362,206 B2
• Filed: 06/28/2005
• Issued: 01/29/2013
• Est. Priority Date: 06/28/2004
• Status: Expired due to Fees

• Current Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Original Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Inventors: Wallach, David, Appel, Elena, Goncahrov, Tanya
• Primary Examiner(s): ROMEO, DAVID S
• Application Number:
  US11/629,258
• Publication Number:
  US 20110123480A1
• Time in Patent Office: 2,772 Days
• Field of Search: None
• US Class Current: 530/350
• 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 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/06 for tuberculosis
• CPC Class Codes: A61P 33/06 Antimalarials
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• 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 14/21 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 14/7151 : for tumor necrosis factor [TNF], for lymphotoxin [LT]
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/55 : containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides a chimeric protein capable of killing or modifying a cell expressing abnormally high levels of a ligand of a receptor of the TNF/NGF family, comprising the amino acid sequence of at least one polypeptide consisting of an extracellular portion of said receptor connected to an effector molecule. In addition the invention provides pharmaceutical compositions comprising said chimeric protein and use thereof.

Claims

1. A chimeric protein comprising the amino acid sequence of at least one polypeptide consisting of an extracellular portion of a TNF receptor, said polypeptide being connected to a Pseudomonas exotoxin fragment, wherein the Pseudomonas exotoxin fragment lacks the cell binding domain and wherein said extracellular portion of the TNF receptor binds TNFα

.

2. The chimeric protein according to claim 1, wherein the protein comprises Pseudomonas exotoxin domains II, Ib and III.

3. The chimeric protein according to claim 2, wherein said fragment of Pseudomonas exotoxin (PE) comprises the amino acid sequence of SEQ ID NO:

4.

4. The chimeric protein according to claim 1, comprising the amino acid sequence of SEQ ID NO:

2.

14. A pharmaceutical composition comprising a chimeric protein according to claim 1 and a pharmaceutically acceptable carrier.

15. The pharmaceutical composition according to claim 14, wherein the chimeric protein is TBP-PE.

5. An isolated DNA sequence encoding a chimeric protein comprising the amino acid sequence of at least one polypeptide consisting of an extracellular portion of a TNF receptor, said polypeptide being connected to a Pseudomonas exotoxin fragment, wherein the Pseudomonas exotoxin fragment lacks the cell binding domain and wherein said extracellular portion of the TNF receptor binds TNFα

.

6. The DNA sequence according to claim 5, further encoding a signal peptide for secretion in eukaryotic cells.

7. The DNA according to claim 5, comprising the nucleotide sequence of SEQ ID NO:

1.

8. An expression vector comprising a DNA sequence according to claim 5.

9. A host cell comprising an expression vector according to claim 8.

10. The host cell according to claim 9, wherein the cell is an eukaryotic cell.

11. The host cell according to claim 10, wherein the eukaryotic cell is selected from the group consisting of HeLa, CHO, HEK293, THPI, Yeast, and insect cells.

12. The host cell according to claim 9, wherein the cell is a prokaryotic cell.

13. A method for producing a chimeric protein, comprising culturing a host cell according to any one of claims 9 to 12 and isolating the chimeric protein produced.

16. A composition comprising a DNA according to claim 5 and a carrier.

17. A composition comprising a vector according to claim 8 and a carrier.