Production and delivery of a stable collagen

Patent metadata

Field	Value
Publication number	US-9040484-B2
Application number	US-201214112444-A
Country	US
Kind code	B2
Filing date	Apr 26, 2012
Priority date	Apr 26, 2011
Publication date	May 26, 2015
Grant date	May 26, 2015

Abstract

Improved methods are provided for the recombinant synthesis of collagen, particularly collagen VII, in host cell, and for therapeutic delivery of the same. The recombinant collagen is produced in a host cell that has increased levels of prolyl-4-hydroxylase, relative to basal cell levels. The collagen produced by the methods of the invention has increased numbers of modified proline residues, relative to a recombinant collagen produced in a host cell having basal levels of prolyl-4-hydroxylase. The increased proline modification provides for a collagen having increased stability, including increased in vivo stability.

First claim

What is claimed is:

1. A method of synthesizing collagen VII, the method comprising:
synthesizing recombinant collagen VII in a host cell that has been genetically modified to increase expression of prolyl-4-hydroxylase, wherein the genetic modification to increase expression of prolyl-4-hydroxylase comprises (i) introduction into the cell of an episomal vector comprising a genetic sequence encoding prolyl-4-hydroxylase operably linked to a promoter, or (ii) modifying the genomic sequence of an endogenous prolyl-4-hydroxylase in said cell to increase expression;
wherein recombinantly produced collagen VII has increased stability relative to a collagen produced in a comparable host cell lacking said genetic modification.

2. The method of claim 1 , wherein said episomal vector further comprises a genetic sequence encoding collagen VII operably linked to a promoter.

3. The method of claim 1 , wherein one or both of said collagen VII and said prolyl-4-hydroxylase are human.

4. The method of claim 1 , wherein said host cell is a mammalian cell.

5. The method of claim 1 , further comprising the step of isolating said collagen VII from said cell.

6. The method of claim 5 , further comprising the step of administering said collagen VII to an individual in need thereof.

7. The method of claim 6 , wherein said administration is intradermal.

8. The method of claim 6 , wherein said individual suffers from epidermolysis bullosa.

9. The method of claim 8 , wherein said epidermolysis bullosa is the result of a genetic defect in collagen VII.

Assignees

• Marinkovich M Peter
• Lane Alfred T
• Rajadas Jayakumar
• Univ Leland Stanford Junior
• Us Dept Veterans Affairs

Inventors

• Marinkovich M Peter
• Lane Alfred T
• Rajadas Jayakumar

Classifications

• C07K14/78 Primary

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title

• A61M2037/0023

  Drug applicators using microneedles · CPC title

• A61M37/0015 Primary

  by using microneedles · CPC title

• C12N9/0071

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title

• C12N15/67

  General methods for enhancing the expression · CPC title