# US 10,441,634 B2 · Treatment of peripheral vascular disease using protein solutions

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,441,634 B2
- Document ID: US10441634B2
- Status: Granted
- Kind: B2
- Application number: 15/836,249
- Filed: 2017-12-08
- Published: 2018-04-12
- Granted: 2019-10-15
- Earliest claimed: 2013-03-15
- Assignees: Biomet Biologics, LLC
- Applicants: Biomet Biologics, LLC
- Inventors: Matthew D. Landrigan; Krista Toler; Jennifer E. Woodell-May; David L. Suter
- Web page: https://patentgrep.com/patent/US10441634B2

## CPC

- A61K 38/19: Cytokines; Lymphokines; Interferons
- A61K 35/14: Blood; Artificial blood (perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42)
- A61K 35/15: Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells (presenting a specific antigen A61K39/00; therapeutic combinations of antibodies, or fragments thereof, and blood-derived cells A61K39/00)
- A61K 35/19: Platelets; Megacaryocytes
- A61K 38/1703: {from vertebrates}
- A61K 38/1793: {for cytokines; for lymphokines; for interferons}
- A61K 38/1808: {Epidermal growth factor [EGF] urogastrone}
- A61K 38/1858: {Platelet-derived growth factor [PDGF]}
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Methods of treating peripheral vascular disease comprising administering a protein solution site of a defect at least two proteins from the group IL-1ra, sTNF-RI, sTNF-RII, IGF-I, EGF, HGF, PDGF-AB, PDGF-BB, VEGF, TGF-β1, and sIL-1RII. The solution may also comprise white blood cells, platelets, concentrated bone marrow aspirate, and combinations thereof.

## Claims

### Claim 1

A method for improving arterial blood flow in an occluded artery, comprising: concentrating whole blood, a fraction of whole blood, white blood cells, bone marrow, a fraction of bone marrow, adipose tissue, a fraction of adipose tissue, or a combination of any of the foregoing, obtained from the subject to form an autologous therapeutic composition; and administering the therapeutic composition at or near the ischemic tissue, the therapeutic composition comprising at least about 10,000 pg/ml interleukin-1 receptor antagonist (IL-ira), at least about 15,000 pg/ml soluble interkeukin-1 receptor II (sIL-iRII), and at least two proteins selected from at least about 1,200 pg/ml soluble tumor necrosis factor receptor I (sTNF-RI), at least about 3,000 pg/ml soluble tumor necrosis factor receptor II (sTNF-RII), at least about 130,000 pg/ml insulin-like growth factor-I (IGF-I), at least about 800 pg/ml epidermal growth factor (EGF), at least about 1000 pg/ml hepatocyte growth factor (HGF), at least about 35,000 pg/ml platelet-derived growth factor-AB (PDGF-AB), at least about 10,000 pg/ml PDGF-BB platelet-derived growth factor-BB (PDGF-BB), at least about 500 pg/ml vascular endothelial growth factor (VEGF), and at least about 100,000 pg/ml transforming growth factor-β1 (TGF-β1), wherein the concentration of each selected protein in the therapeutic composition is greater than the concentration of the selected protein in the biological material.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the therapeutic composition further comprises the subject's white blood cells at a concentration of at least 15,000/μl.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the therapeutic composition further comprises
the subject's platelets at a concentration of at least 400,000/μl.

### Claim 4

A method for improving arterial blood flow in an occluded artery, comprising topically injecting at or near the site of the ischemic tissue an autologous composition comprising: at least 10,000 pg/ml interleukin-1 receptor antagonist (IL-1ra); at least 15,000 pg/ml soluble interleukin-1 receptor II (sIL-IRII); and at least 1,000 pg/ml hepatocyte growth factor (HGF), wherein the IL-ira, the sTNF-R1 and the sIL-1RII are prepared from whole blood, a fraction of whole blood, bone marrow, a fraction of bone marrow, adipose tissue, a fraction of adipose tissue, or a combination of any of the foregoing.

### Claim 5 (depends on 4)

The method according to claim 4, wherein the composition further comprises at least about 1,200 pg/ml soluble tumor necrosis factor receptor I (sTNF-RI).

### Claim 6 (depends on 4)

The method according to claim 4, wherein the composition further comprises a protein selected from the group consisting of soluble tumor necrosis factor-receptor II (sTNF-RII), insulin-like growth factor-I (IGF-1), epidermal growth factor (EGF), hepatocyte growth factor (HGF), platelet-derived growth factor-AB (PDGF-AB), platelet-derived growth factor-BB (PDGF-BB), vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), and sIL-1RII, and mixtures of any of the foregoing, wherein the concentration of the selected protein in the composition is greater than the concentration of the protein in the subject's blood.

### Claim 7 (depends on 4)

The method according to claim 4, wherein the composition further comprises concentrated bone marrow aspirate.

### Claim 8

A method for improving arterial blood flow in an occluded artery, comprising: obtaining a cytokine cell suspension from the subject; processing the cytokine-cell suspension to produce an autologous protein solution comprising at least 10,000 pg/ml interleukin-1 receptor antagonist (IL-1ra) and at least 15,000 pg/ml soluble interleukin-1 receptor II (sIL-IRII), and hepatocyte growth factor at a concentration of at least about 1,000 pg/ml wherein the ratio of IL-Ira to interleukin-β1 in the composition is greater than 1000, and the ratio of sIL-1RII to IL-βI in the composition is greater than 1; and
administering the autologous protein solution by topical injection at or near the site of the tissue ischemia.

### Claim 9 (depends on 8)

The method according to claim 8 wherein the cytokine cell suspension comprises whole blood, bone marrow aspirate, adipose tissue, urine, a fraction of any one of the foregoing, and a mixture of any of the foregoing.

### Claim 10 (depends on 8)

The method according to claim 8, wherein processing comprises separating blood to create a platelet-rich plasma fraction comprising white blood cells at a concentration of at least 15,000/μl.

### Claim 11 (depends on 8)

The method according to claim 8, wherein processing includes contacting the cytokine-cell suspension with polyacrylamide beads to form the autologous protein solution.

### Claim 12 (depends on 8)

The method according to claim 8, wherein processing includes contacting the cytokine-cell suspension with a solid extraction material.

### Claim 13 (depends on 12)

The method according to claim 12, wherein the solid extraction material is selected from the group consisting of corundum, quartz, titanium, dextran, agarose, polyacrylamide, polystyrene, polyethylene, polyvinyl chloride, polypropylene, and combinations thereof.

### Claim 14 (depends on 13)

The method according to claim 13, wherein the solid extraction material comprises a form selected from the group consisting of a bead, fiber, powder, porous material, and combinations thereof.

### Claim 15 (depends on 12)

The method according to claim 12, wherein the solid extraction material comprises a desiccating material.

### Claim 16 (depends on 8)

The method according to claim 8, further comprising:
obtaining bone marrow aspirate from the subject;
concentrating the bone marrow aspirate to produce a concentrated bone marrow aspirate (cBMA);
administering the cBMA to the subject.

### Claim 17 (depends on 16)

The method according to claim 16, wherein the autologous protein solution further comprises soluble tumor necrosis factor receptor II at a concentration of at least about 3,000 pg/ml.

### Claim 18 (depends on 8)

The method according to claim 8, wherein the autologous protein solution comprises white blood cells at a concentration of at least about 30,000/μl.

### Claim 19 (depends on 8)

The method according to claim 8, wherein treating tissue ischemia includes healing a lesion at or near the site of the tissue ischemia.