FUSION PROTEINS FORMING TRIMERS

• US 20150126710A1
• Filed: 07/02/2014
• Published: 05/07/2015
• Est. Priority Date: 01/09/2009
• Status: Abandoned Application

• Current Assignee: Apogenix AG
• Original Assignee: Apogenix Gmbh
• Inventors: HILL, Oliver, Branschdel, Marcus, Gieffers, Christian, Thiemann, Meinolf
• Application Number:
  US14/322,852
• Publication Number:
  US 20150126710A1
• Time in Patent Office: Days
• US Class Current: 530/351
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/4726 Lectins
• 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 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/21 containing a His-tag
• CPC Class Codes: C07K 2319/22 containing a Strep-tag
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

The present invention refers to fusion proteins comprising a neck region and carbohydrate recognition domain of a collectin trimerization domain, a linker element and an effector polypeptide. Further the invention refers to a nucleic acid encoding the said fusion protein. The fusion proteins, the nucleic acid, and the cell are suitable as pharmaceutical composition or for therapeutic, diagnostic and/or research applications as described herein.

Claims

1. A fusion protein comprising

(i) a collectin family trimerization domain selected from the group consisting of the amino acid residues 217-375, 218-375, 219-375, 220-375, 221-375, 222-375, 223-375, 224-375, and 225-375 of SEQ ID NO:

21, optionally further consisting of an amino acid substitution at residue 355;

(ii) a linker element; and

(iii) TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) or a receptor binding domain thereof;

wherein said linker element has 25 or less amino acids.

2. The fusion protein claim 1, wherein (iii) is located N-terminally of the collectin family neck region.

3. The fusion protein of claim 2, wherein the linker element consists of amino acids glycine and serine.

4. The fusion protein of claim 2, wherein the linker has the amino acid sequence of SEQ ID NO:

33, which is (GSS)a(SSG)b(GSG)c, wherein a, b, c is each independently 0, 1, 2, 3, 4, 5 or 6 such that the linker has 25 or less amino acid.

5. The fusion protein of claim 2, wherein the linker element has the amino acid sequence of SEQ ID NO:

56, which is GSSGSSGSSGSG.

6. The fusion protein of claim 2, wherein the linker element has the amino acid sequence of SEQ ID NO:

34, which is GSSGSSGSSGS.

7. The fusion protein of claim 2, further comprising an N-terminal signal peptide domain.

8. The fusion protein of claim 2, further comprising a recognition/purification domain selected from the group consisting of a strep-tag domain and a poly-His domain.

9. The fusion protein of claim 1, wherein the amino acid residue 355 in SEQ ID NO:

21 is optionally substituted to a polar amino acid selected from the group consisting of aspartic acid, glutamic acid, asparagine and glutamine.

10. The fusion protein of claim 2, wherein (iii) comprises the sequence of human TRAIL, which is SEQ ID NO:

10.

11. The fusion protein of claim 2, wherein (iii) comprises amino acids 95-281, 116-281, 117-281, 118-281, 119-281 or 120-281 of human TRAIL (SEQ ID NO:

10).

12. A trimeric complex consisting of three identical fusion proteins of claim 1.