Method of producing concentrated non-buffered solutions of fibronectin

• US 6,528,483 B2
• Filed: 08/21/1999
• Issued: 03/04/2003
• Est. Priority Date: 06/07/1995
• Status: Expired due to Fees

• Current Assignee: Andre Beaulieu
• Original Assignee: Andre Beaulieu
• Inventors: Paquin, Robert, Beaulieu, André
• Primary Examiner(s): KRASS, FREDERICK F
• Application Number:
  US09/331,344
• Publication Number:
  US 20020010130A1
• Time in Patent Office: 1,291 Days
• Field of Search: 514/8, 530/380, 530/395, 424/78.06
• US Class Current: 514/9.3
• CPC Class Codes: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/32 Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers , poly(meth)acrylates, or polyvinyl pyrrolidone
• CPC Class Codes: A61K 47/38 Cellulose; Derivatives thereof
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00

Abstract

Aqueous, non-buffered solutions containing high concentrations of fibronectin and other wound healing promoters are produced, which can then be used to make topical gel and cream formulations with high concentrations of these wound healing promoters. The formulations are used for the healing of cutaneous wounds. The formulations provide slow release and increased contact time of fibronectin and of other wound healing promoters to the wound site leading to effective absorption of the promoters.

Claims

1. A method of preparing a non-buffered aqueous solution containing at least 2 mg/ml of fibronectin comprising the step of adding purified fibronectin to basic water.

2. The method of claim 1 wherein the water is demineralized water.

3. The method of claim 1 wherein the fibronectin is human plasma fibronectin.

4. The method of claim 1 wherein the purified fibronectin as lyophilized.

5. A method of preparing a non-buffered aqueous solution containing at least 2 mg/ml of fibronectin comprising the steps of:

a) adjusting the pH of deminealized water from about 8.0 to about 11.0 and

b) adding sufficient purified fibronectin to the water of step a to reach a concentration of at least 2 mg/ml of fibronectin.

6. The method of claim 5, wherein the non-buffered aqueous solution contains about 2 mg/ml to about 10 mg/ml of fibronectin.

7. The method of claim 5, wherein the fibronectin is human plasma fibronectin.

8. The method of claim 5, wherein the purified fibronectin is lyophilized.

9. A method of preparing a non-buffered aqueous solution containing at least 2 mg/ml of fibronectin comprising the steps of:

a) adjusting the pH of demineralized water to about 9.0 and

b) adding sufficient purified fibronectin to the water of step a to reach a concentration of at least 2 mg/ml of fibronectin.

10. A non-buffered aqueous solution containing more than 5 mg/ml to about 10 mg/ml of solubilized fibronectin.

11. A pharmaceutical formulation comprising more than 0.5% to about 1.0% by weight fibronectin based on the total weight of the pharmaceutical formulation comprising:

a) a non-buffered, aqueous fibronectin solution containing more than 5 mg/ml to about 10 mg/ml of solubilized fibronectin, and

b) a pharmaceutically acceptable carrier.

12. The pharmaceutical formulation of claim 11, wherein the pharmaceutically acceptable carrier is a water soluble, pharmaceutical polymer having a viscosity of 50,000 to about 1,000,000 cps at room temperature.

13. The pharmaceutical formulation of claim 11, further comprising at least one extracellular matrix protein selected from the group consisting of thrombospondin, laminin, vitronectin, and fibrinogen.

14. The pharmaceutical formulation of claim 11, further comprising at least one growth factor.

15. An aqueous gel formulation for healing chronic wounds comprising about 0.28% carbomer and more than 0.5% to about 1.0% by weight of fibronectin, based on the total weight of the gel formulation.