Stable macroscopic membranes formed by self-assembly of amphiphilic peptides and uses therefor

• US 7,098,028 B2
• Filed: 03/17/2003
• Issued: 08/29/2006
• Est. Priority Date: 12/28/1992
• Status: Expired due to Fees

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Holmes, Todd, DiPersio, C. Michael, Zhang, Shuguang, Lockshin, Curtis, Rich, Alexander
• Primary Examiner(s): Weber, Jon
• Assistant Examiner(s): TSAY, MARSHA M
• Application Number:
  US10/390,472
• Publication Number:
  US 20040087013A1
• Time in Patent Office: 1,261 Days
• Field of Search: None
• US Class Current: 435/401
• CPC Class Codes: A61K 9/009 Sachets, pouches characterised by the material or function of the envelope with gastric retention A61K9/0065; sachets which are not administered but function merely as a container are classified according to the content, e.g. sachets comprising powder for reconstitution of a drink A61K9/0095
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/395 from Saccharomyces
• CPC Class Codes: C07K 5/0815 with the first amino acid being basic
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Described herein is the self-assembly of amphiphilic peptides, i.e., peptides with alternating hydrophobic and hydrophilic residues, into macroscopic membranes. The membrane-forming peptides are greater than 12 amino acids in length, and preferably at least 16 amino acids, are complementary and are structurally compatible. Specifically, two peptides, (AEAEAKAK)₂ (ARARADAD)₂, were shown to self-assemble into macroscopic membranes. Conditions under which the peptides self-assemble into macroscopic membranes and methods for producing the membranes are also described. The macroscopic membranes have several interesting properties: they are stable in aqueous solution, serum, and ethanol, are highly resistant to heat, alkaline and acidic pH, chemical denaturants, and proteolytic digestion, and are non-cytotoxic. The membranes are potentially useful in biomaterial applications such as slow-diffusion drug delivery systems, artificial skin, and separation matrices, and as experimental models for Alzheimer'"'"'s disease and scrapie infection. The sequence of the peptide, EAK16, was derived from a putative Z-DNA binding protein from yeast, called zuotin. The cloning and characterization of the ZUO1 gene are also described.

Claims

1. A drug delivery system comprising:

membranes formed by self-assembly of amphiphilic peptides, wherein the peptides have alternating hydrophobic and hydrophilic amino acids, are complementary and structurally compatible, and self-assemble in the presence of a monovalent cation; and

;

a drug.

2. The drug delivery system of claim 1, wherein the drug is a protein.

3. The drug delivery system of claim 2, wherein the protein is selected from the group consisting of:

erythropoietin, tissue type plasminogen activator, synthetic hemoglobin and insulin.

4. The drug delivery system of claim 1, wherein the hydrophilic amino acids are acidic and basic amino acids.

5. The drug delivery system of claim 4, wherein the acidic amino acids are independently selected from the group consisting of aspartic acid and glutamic acid.

6. The drug delivery system of claim 4, wherein the basic amino acids are independently selected from the group consisting of arginine, lysine, histidine and ornithine.

7. The drug delivery system of claim 1, wherein the hydrophobic amino acids are selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tyrosine, tryptophan, and glycine.

8. A method of delivering a drug to a subject comprising:

administering the drug delivery system of claim 1 to the subject.

9. A cell culture system comprising:

membranes formed by self-assembly of amphiphilic peptides, wherein the peptides have alternating hydrophobic and hydrophilic amino acids, are complementary and structurally compatible; and

a cell culture vessel.

10. The cell culture system of claim 9, wherein the membranes are stacked in the cell culture vessel.

11. The cell culture system of claim 9, further comprising cells that adhere to surfaces of the membranes.

12. The cell culture system of claim 11, wherein the cells are engineered to produce a product.

13. The cell culture system of claim 9, wherein the hydrophilic amino acids are acidic and basic amino acids.

14. The cell culture system of claim 13, wherein the acidic amino acids are independently selected from the group consisting of aspartic acid and glutamic acid.

15. The cell culture system of claim 13, wherein the basic amino acids are independently selected from the group consisting of arginine, lysine, histidine and ornithine.

16. The cell culture system of claim 9, wherein the hydrophobic amino acids are selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tyrosine, tryptophan, and glycine.

17. A method of culturing cells comprising steps of:

stacking membranes formed by self-assembly of amphiphilic peptides, wherein the peptides have alternating hydrophobic and hydrophilic amino acids and are complementary and structurally compatible, in a tissue culture vessel in the presence of culture medium, wherein cells adhere to the surfaces of the membranes; and

maintaining the cells under conditions suitable for growth.

18. The method of claim 17, wherein the cells are engineered to produce a product.

19. The method of claim 18, further comprising the step of:

harvesting the product.

20. A product produced according to the method of claim 19.

21. A method of treating a subject comprising the step of:

introducing a composition comprising amphiphilic peptides, wherein the peptides have alternating hydrophobic and hydrophilic amino acids, are complementary and structurally compatible, and self-assemble to form a beta-sheet structure in the presence of a monovalent cation, into the subject.

22. The method of claim 21, wherein the composition comprises a drug.

23. The method of claim 21, wherein the step of introducing the composition comprises injecting the composition into the subject.