Matrix scaffold with antimicrobial activity

• US 9,931,437 B2
• Filed: 02/04/2015
• Issued: 04/03/2018
• Est. Priority Date: 06/26/2012
• Status: Active Grant

• Current Assignee: Worcester Polytechnic Institute
• Original Assignee: Worcester Polytechnic Institute
• Inventors: Rolle, Marsha, Prifti, Fioleda, Malcuit, Christopher, Camesano, Terri Anne, Dominko, Tanja, Kole, Denis
• Primary Examiner(s): Alstrum-Acevedo, James H
• Assistant Examiner(s): Martinez, Tara L
• Application Number:
  US14/613,767
• Publication Number:
  US 20150202350A1
• Time in Patent Office: 1,154 Days
• Field of Search: None
• CPC Class Codes: A61L 2300/404 Biocides, antimicrobial agents, antiseptic agents
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: C07K 14/4723 Cationic antimicrobial peptides, e.g. defensins
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

The invention provides a scaffold of extracellular matrix polymers with recombinant chimeric peptides tethered thereto. The invention also provides recombinant chimeric peptides of antimicrobial peptides and extracellular matrix binding domains. The invention also provides methods for treating chronic wounds using the scaffold and/or recombinant chimeric peptides.

Claims

1. A scaffold comprising one or more extracellular matrix polymers and one or more chimeric peptides comprising one or more antimicrobial peptides and one or more extracellular matrix binding domains, wherein said one or more antimicrobial peptides is MKT QRDGHSLGRWSL VLLLLGLVMPL AII AQ VLS YKE AVLRAIDGINQRS SD ANL YRLL DLDPRPTMDGDPDTPKPVSFTVKETVCPRTTQQSPEDCDFKKDGLVKRCMGTVTLNQA RGSFDISCDKDNKRFALLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES (SEQ ID NO:

4) and said extracellular matrix binding domain is linked to the C-terminal end of SEQ ID NO;

4.

2. The scaffold according to claim 1, wherein said scaffold is produced by transfected cells.

3. The scaffold according to claim 2, wherein said transfected cells are removed from said produced scaffold.

4. The scaffold according to claim 2, wherein said transfected cells are selected from the group consisting of fibroblasts, keratinocytes, human lung carcinoma cells, smooth muscle cells, skeletal muscle cells, stem cells and combinations thereof.

5. The scaffold according to claim 1, wherein said one or more chimeric peptides are added to previously produced extracellular matrix polymers.

6. The scaffold according to claim 1, wherein said one or more extracellular matrix polymers are combined with one or more artificial polymers.

7. The scaffold according to claim 6, wherein said one or more extracellular matrix polymer is selected from the group consisting of collagen, fibronectin, laminin, elastin, hyaluronan, fibrin, gelatin, alginate, glycosaminoglycans and combinations thereof.

8. The scaffold according to claim 1, wherein said one or more extracellular matrix binding domains is selected from the group consisting of collagen binding domain, fibronectin binding domain, laminin binding domain, elastin binding domain, hyaluronan binding domain, fibrin binding domain, gelatin binding domain, alginate binding domain, glycosaminoglycan binding domain and combinations thereof.

9. The scaffold according to claim 1, wherein said one or more extracellular matrix binding domains is selected from the group consisting of TKKTLRT (SEQ ID NO:

5), CQDSETGTFY (SEQ ID NO;

     6), and combinations thereof.

11. The scaffold according to claim 1, wherein the scaffold is produced by a combination of transfected cells and non-transfected fibroblasts.

12. The scaffold according to claim 1, wherein said one or more chimeric peptides further comprises an intervening peptide domain between said antimicrobial peptide and said extracellular matrix binding domain.

13. The scaffold according to claim 1, wherein said one or more chimeric peptides are produced by transfected cells.

14. The scaffold according to claim 11, wherein said transfected cells are selected from the group consisting of fibroblasts, keratinocytes, human lung carcinoma cells, smooth muscle cells, skeletal muscle cells, stem cells and combinations thereof.

15. The scaffold according to claim 12, wherein said intervening peptide domain comprises five aspartic acid residues, two lysine residues and a tyrosine residue.

10. A scaffold comprising one or more extracellular matrix polymers and one or more chimeric peptides comprising one or more antimicrobial peptides and one or more extracellular matrix binding domains, wherein said one or more chimeric peptide comprises MKTQRDGHSLGRWSLVLLLLGLVMPLAIIAQVLSYKEAVLRAIDGINQRSSDANLYR LL DLDPRPTMDGDPDTPKPVSFTVKETVCPRTTQQSPEDCDFKKDGLVKRCMGTVTLNQA R GSFDISCDKDNKRFALLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES (SEQ ID NO:

4) and one or more of TKKTLRT (SEQ ID NO;

     5) or CQDSETGTFY (SEQ ID NO;

     6), and wherein said extracellular matrix binding domain is linked to the C-terminal end of SEQ ID NO;

4.

16. The scaffold according to claim 10, wherein said scaffold is produced by transfected cells.

17. The scaffold according to claim 16, wherein said transfected cells are removed from said produced scaffold.

18. The scaffold according to claim 16, wherein said transfected cells are selected from the group consisting of fibroblasts, keratinocytes, human lung carcinoma cells, smooth muscle cells, skeletal muscle cells, stem cells and combinations thereof.

19. The scaffold according to claim 10, wherein said one or more chimeric peptides are added to previously produced extracellular matrix polymers.

20. The scaffold according to claim 10, wherein said one or more extracellular matrix polymers are combined with one or more artificial polymers.

21. The scaffold according to claim 20, wherein said one or more extracellular matrix polymer is selected from the group consisting of collagen, fibronectin, laminin, elastin, hyaluronan, fibrin, gelatin, alginate, glycosaminoglycans and combinations thereof.

22. The scaffold according to claim 10, wherein the scaffold is produced by a combination of transfected cells and non-transfected fibroblasts.

23. The scaffold according to claim 10, wherein said one or more chimeric peptides further comprises an intervening peptide domain between said antimicrobial peptide and said extracellular matrix binding domain.

24. The scaffold according to claim 10, wherein said one or more chimeric peptides are produced by transfected cells.

25. The scaffold according to claim 22, wherein said transfected cells are selected from the group consisting of fibroblasts, keratinocytes, human lung carcinoma cells, smooth muscle cells, skeletal muscle cells, stem cells and combinations thereof.

26. The scaffold according to claim 23, wherein said intervening peptide domain comprises five aspartic acid residues, two lysine residues and a tyrosine residue.