Methods and compositions for wound treatment

Patent metadata

Field	Value
Publication number	US-10101339-B2
Application number	US-201615191632-A
Country	US
Kind code	B2
Filing date	Jun 24, 2016
Priority date	Dec 21, 2010
Publication date	Oct 16, 2018
Grant date	Oct 16, 2018

Abstract

The present disclosure relates to methods for identifying proteins or peptide motifs of intracellular, extracellular, or extracellular matrix proteins specifically exposed in wound sites, as well as compositions for treating wounds, and methods for their use.

First claim

What is claimed is:

1. A method for identifying a wound target for facilitating delivery of a hemostatic agent to a wound, wherein the wound target is an exposed protein, peptide motif or epitope, wherein the exposed protein, peptide motif or epitope becomes exposed upon wounding, the method comprising:
removing predetermined internal organ tissue from a mammalian subject, the predetermined internal organ tissue being selected from the group consisting of esophagus, stomach, small and large intestines, pancreas, liver, kidney, bladder, urethra, heart, lung, brain, and reproductive organs;
wounding the removed predetermined internal organ tissue to expose one or more proteins, peptide motifs or epitopes, wherein the one or more proteins, peptide motifs or epitopes are not exposed in unwounded predetermined internal organ tissue and further wherein the one or more proteins, peptide motifs or epitopes are selected from the group consisting of actin, and a cryptic tripeptide arginine-glycine-aspartic acid (RGD) epitope;
incubating the wounded predetermined internal organ tissue with a preselected detectable wound targeting moiety;
washing away unbound wound targeting moiety from the wounded predetermined internal organ tissue after a predetermined incubation period;
detecting whether the wound targeting moiety is specifically bound to an exposed protein, peptide motif or epitope; and
identifying the exposed protein, peptide motif or epitope specifically bound to the wound targeting moiety upon wounding as the wound target; wherein the detectable wound targeting moiety is a detectable antibody specifically binding to said actin or said RGD epitope.

2. The method of claim 1 , wherein the wound targeting moiety is a fluorescently labeled antibody.

3. The method of claim 1 , wherein the antibody is an antibody to actin protein.

4. The method of claim 1 , wherein identifying the exposed protein, peptide motif or epitope specifically bound upon wounding as a wound target further includes determining that the wound targeting moiety does not bind appreciably above predetermined background levels to unwounded tissue.

5. The method of claim 1 , wherein the exposed protein, peptide motif or epitope is a cryptic tripeptide arginine-glycine-aspartic acid (RGD) epitope.

6. A method for identifying a wound target for facilitating delivery of a hemostatic agent to a wound, wherein the wound target is an exposed protein, peptide motif or epitope, the method comprising:
removing predetermined internal organ tissue from a mammalian subject, the predetermined internal organ tissue being selected from the group consisting of esophagus, stomach, small and large intestines, pancreas, liver, kidney, bladder, urethra, heart, lung, brain, and reproductive organs;
wounding the removed predetermined internal organ tissue to expose a first protein, peptide motif or epitope, wherein the first protein, peptide motif or epitope is not exposed in unwounded predetermined internal organ tissue and further wherein the first protein, peptide motif or epitope is selected from the group consisting of actin, and a cryptic tripeptide arginine-glycine-aspartic acid (RGD) epitope;
incubating the wounded predetermined internal organ tissue with a first preselected detectable wound targeting moiety; wherein the first preselected detectable wound targeting moiety is a detectable antibody specifically binding to said actin or said RGD epitope
washing away unbound detectable wound targeting moiety from the wounded predetermined internal organ tissue after a predetermined incubation period;
detecting whether the detectable wound targeting moiety is specifically bound to the first exposed protein, peptide motif or epitope;
determining that the detectable wound targeting moiety does not bind appreciably above predetermined background levels to unwounded tissue;
identifying the first exposed protein, peptide motif or epitope specifically bound to the first detectable wound targeting moiety as a first wound target;
identifying an amino acid sequence of the identified first exposed protein, peptide motif or epitope and selecting a second protein, peptide motif or epitope having a similar amino acid sequence to the first protein, peptide motif or epitope as a candidate wound target and performing the incubating, washing, detecting and determining steps above to confirm the second protein, peptide motif or epitope as a second wound target.

7. The method of claim 6 , wherein the first preselected detectable wound targeting moiety is a detectable antibody specific for the actin.

8. The method of claim 1 , wherein the exposed protein, peptide motif or epitope is a cryptic tripeptide arginine-glycine-aspartic acid (RGD) epitope.

Assignees

• Leidos Inc

Inventors

• Dresios John
• Griffey Richard

Classifications

• C07K16/18

  against material from animals or humans · CPC title

• A61K38/1777

  Integrin superfamily · CPC title

• G01N33/6893

  related to diseases not provided for elsewhere · CPC title

• A61K47/6811

  the drug being a protein or peptide, e.g. transferrin or bleomycin · CPC title

• G01N2333/78

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