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Details for Patent: 4,849,407

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Details for Patent: 4,849,407

Title: Biologically active mosaic proteins
Abstract:Dimeric proteins having substantially the same biological activity as PDGF are disclosed. More specifically, the proteins may have two polypeptide chains, one of the chains being a mosaic of amino acid sequences substantially identical to portions of the A- or B-chains of PDGF, the second of the chains being substantially homologous to either the A-chain or the B-chain of PDGF, the proteins being chemotactic or mitogenic for fibroblasts. Alternatively, each of the two polypeptide chains may be a mosaic of amino acid sequences as described above. Therapeutic compositions containing these proteins and methods for enhancing the wound-healing process in warm-blooded animals are also disclosed.
Inventor(s): Murray; Mark J. (Seattle, WA), Kelly; James D. (Seattle, WA)
Assignee: ZymoGenetics, Inc. (Seattle, WA)
Filing Date:Dec 15, 1986
Application Number:06/941,970
Claims:1. A protein having two polypeptide chains, one of said chains being a mosaic of amino acid sequences substantially identical to portions of the A- or B-chains of PDGF, the second of said chains being substantially homologous to the A-chain of PDFG, said protein being chemotactic or mitogenic for fibroblasts, wherein said mosaic chain comprises a sequence selected from the group consisting of:

(a) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-109; and

(b) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-97 fused in reading frame to A-chain amino acids 92-104;

and wherein said second chain comprises a sequence selected from the group consisting of:

(a) the amino acid sequence of the A-chain of PDGF from amino acid 15 to amino acid 104;

(b) the amino acid sequence of the A-chain of PDGF from amino acid 23 to amino acid 104;

(c) the amino acid sequence of the A-chain of PDGF from amino acid 9 to amino acid 95;

(d) the amino acid sequence of the A-chain of PDGF from amino acid 23 to amino acid 95;

(e) the amino acid sequence of the A-chain of PDGF from amino acid 1 to amino acid 95; and

(f) the amino acid sequence of the A-chain of PDGF from amino acid 1 to amino acid 104.

2. A protein having two polypeptide chains, one of said chains being a mosaic of amino acid sequences substantially identical to portions of the A- or B-chains of PDGF, the second of said chains being substantially homologous to the B-chain of PDGF, said protein being chemotactic or mitogenic for fibroblasts, wherein said mosaic chain comprises a sequence selected from the group consisting of:

(a) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-109; and

(b) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-97 fused in reading frame to A-chain amino acids 92-104;

and wherein said second chain comprises a sequence selected from the group consisting of:

(a) the amino acid sequence of the B-chain of PDGF from amino acid 15 to amino acid 109;

(b) the amino acid sequence of the B-chain of PDGF from amino acid 29 to amino acid 109;

(c) the amino acid sequence of the B-chain of PDGF from amino acid 15 to amino acid 101;

(d) the amino acid sequence of the B-chain of PDGF from amino acid 29 to amino acid 101;

(e) the amino acid sequence of the B-chain of PDGF from amino acid 1 to amino acid 101; and

(f) the amino acid sequence of the B-chain of PDGF from amino acid 1 to amino acid 109.

3. A protein having two polypeptide chains, each of said chains being a mosaic of amino acid sequences selected from the group consisting of:

(a) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-109; and

(b) A-chain amino acids 1-17 fused in reading frame to B-chain amino acids 24-97 fused in reading frame to A-chain amino acids 92-194.

4. A wound-healing composition comprising a protein according to claim 1, in combination with a physiologically acceptable carrier or diluent.

5. A wound-healing composition comprising a protein according to claim 2, in combination with a physiologically acceptable carrier or diluent.

6. A wound-healing composition comprising a protein according to claim 3, in combination with a physiologically acceptable carrier or diluent.

7. The wound-healing composition of claim 4, 5 or 6 wherein the carrier or diluent is selected from the group consisting of albumin, saline and sterile water.

8. The wound-healing composition of claim 4, 5 or 6, including an adjuvant.

9. The wound-healing composition of claim 8 wherein the adjuvant is selected from the group consisting of collagen, hyaluronic acid preparations, fibronectin and factor XIII.

10. A method for enhancing the wound-healing process in warm-blooded animals, comprising administering to a warm-blooded animal an effective amount of a wound-healing composition according to claim 4, 5 or 6.
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