Rage fusion proteins
View Patent ↗Disclosed are RAGE fusion proteins comprising RAGE polypeptide sequences linked to a second, non-RAGE polypeptide. The RAGE fusion protein may utilize a RAGE polypeptide domain comprising a RAGE ligand binding site and an interdomain linker directly linked to an immunoglobulin C H 2 domain. Such fusion proteins may provide specific, high affinity binding to RAGE ligands. Also disclosed is the use of the RAGE fusion proteins as therapeutics for RAGE-mediated pathologies.
1. A fusion protein comprising an amino acid sequence which comprises from N-terminus to C-terminus: (a) a Receptor for Advanced Glycation Endproducts (RAGE) polypeptide which comprises a first RAGE immunoglobulin domain and a first RAGE interdomain linker linked to a second RAGE immunoglobulin domain and a second RAGE interdomain linker, such that the N-terminal amino acid of the first RAGE interdomain linker is linked to the C-terminal amino acid of the first RAGE immunoglobulin domain, the N-terminal amino acid of the second RAGE immunoglobulin domain is linked to the C-terminal amino acid of the first RAGE interdomain linker, and the N-terminal amino acid of the second RAGE interdomain linker is linked to the C-terminal amino acid of the second RAGE immunoglobulin domain, wherein the RAGE polypeptide comprises a RAGE ligand binding site, wherein the second RAGE interdomain linker comprises SEQ ID NO: 22 or a sequence 90% identical thereto, and wherein the C-terminal amino acid of the second RAGE interdomain linker is directly linked to the N-terminal amino acid of (b) a polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin, and wherein the fusion protein does not include an immunoglobulin Fc hinge region.
2. The fusion protein of claim 1 , wherein the RAGE ligand binding site comprises the amino acid sequence as set forth in SEQ ID NO: 9 or a sequence 90% identical thereto, or the amino acid sequence as set forth in SEQ ID NO: 10 or a sequence 90% identical thereto.
3. The fusion protein of claim 1 , wherein the RAGE polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 7 or the amino acid sequence as set forth in SEQ ID NO: 8.
4. The fusion protein of claim 1 , wherein the RAGE polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 13 or the amino acid sequence as set forth in SEQ ID NO: 14.
5. The fusion protein of claim 1 , wherein the RAGE polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 15 or the amino acid sequence as set forth in SEQ ID NO: 16.
6. The fusion protein of claim 1 , wherein the RAGE polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 17 or the amino acid sequence as set forth in SEQ ID NO: 18.
7. The fusion protein of claim 1 , wherein the RAGE polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 19 or the amino acid sequence as set forth in SEQ ID NO: 20.
8. The fusion protein of claim 1 , wherein the amino acid sequence of the RAGE polypeptide is as set forth in SEQ ID NO: 19 or SEQ ID NO: 20.
9. The fusion protein of claim 1 , wherein the amino acid sequence of the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin comprises the amino acid sequence as set forth in SEQ ID NO: 38.
10. The fusion protein of claim 1 , wherein the amino acid sequence of the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is as set forth in SEQ ID NO: 38.
11. The fusion protein of claim 1 , comprising the amino acid sequence set forth in SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO: 34.
12. A fusion protein produced by a method comprising culturing a host cell, the host cell comprising an expression vector comprising a nucleic acid that encodes for the fusion protein of claim 1 .
13. The fusion protein of claim 1 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgA, IgD, IgE, IgG or IgM.
14. The fusion protein of claim 13 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgG.
15. The fusion protein of claim 14 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgG1.
16. A pharmaceutical composition comprising a therapeutically effective amount of a RAGE fusion protein in a pharmaceutical carrier, wherein the RAGE fusion protein comprises an amino acid sequence which comprises from N-terminus to C-terminus: (a) a RAGE polypeptide which comprises a first RAGE immunoglobulin domain and a first RAGE interdomain linker linked to a second RAGE immunoglobulin domain and a second RAGE interdomain linker, such that the N-terminal amino acid of the first RAGE interdomain linker is linked to the C-terminal amino acid of the first RAGE immunoglobulin domain, the N-terminal amino acid of the second RAGE immunoglobulin domain is linked to the C-terminal amino acid of the first RAGE interdomain linker, and the N-terminal amino acid of the second RAGE interdomain linker is linked to the C-terminal amino acid of the second RAGE immunoglobulin domain, wherein the RAGE polypeptide comprises a RAGE ligand binding site, wherein the second RAGE interdomain linker comprises SEQ ID NO: 22 or a sequence 90% identical thereto, and wherein the second RAGE interdomain linker is directly linked to (b) a polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin, and wherein the fusion protein does not include an immunoglobulin Fc hinge region.
17. The pharmaceutical composition of claim 16 , wherein the RAGE ligand binding site comprises the amino acid sequence as set forth in SEQ ID NO: 9 or a sequence 90% identical thereto, or the amino acid sequence as set forth in SEQ ID NO: 10 or a sequence 90% identical thereto.
18. The pharmaceutical composition of claim 16 , wherein the fusion protein comprises the amino acid sequence as set forth in SEQ ID NO: 33 or the amino acid sequence as set forth in SEQ ID NO: 34.
19. The pharmaceutical composition of claim 16 , wherein the amino acid sequence of the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin comprises the amino acid sequence as set forth in SEQ ID NO: 38.
20. The pharmaceutical composition of claim 16 , wherein the amino acid sequence of the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is as set forth in SEQ ID NO: 38.
21. The pharmaceutical composition of claim 16 formulated as an injectable solution.
22. The pharmaceutical composition of claim 16 formulated as a sterile lyophilized powder.
23. The pharmaceutical composition of claim 16 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgA, IgD, IgE, IgG or IgM.
24. The pharmaceutical composition of claim 23 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgG.
25. The pharmaceutical composition of claim 24 , wherein the polypeptide comprising at least a portion of a C H 2 domain of an immunoglobulin and a C H 3 domain of an immunoglobulin is derived from a human IgG1.
26. A fusion protein comprising the amino acid sequence set forth in SEQ ID NO: 33.
27. A fusion protein comprising the amino acid sequence set forth in SEQ ID NO: 34.
28. A fusion protein encodable by the nucleic acid sequence as set forth in SEQ ID NO: 30.