Method of binding collagen recognizing substances to liposomes

• US 5,846,561 A
• Filed: 12/10/1996
• Issued: 12/08/1998
• Est. Priority Date: 02/14/1991
• Status: Expired due to Fees

• Current Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Original Assignee: Baxter International Inc.
• Inventors: Margalit, Rimona
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US08/763,214
• Time in Patent Office: 728 Days
• Field of Search: 264/4.3, 428/402.2, 424/1.21, 424/78.04, 424/78.06, 424/450, 424/428, 424/812, 427/2.14, 436/829, 514/62, 514/801
• US Class Current: 424/450
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

Recognizing substances, including epidermal growth factor, gelatin, collagen and hyaluronic acid, have been covalently bound to liposomal surfaces and utilized to attach liposomes onto a cellular or an extracellular matrix (ECM) target site. These "bioadhesive" liposomes offer several advantages including the mutual protection of both the drug and biological environment; an increase in drug bioavailability and retention at the target site; and improved adherence or adhesion to the designated target site.

Claims

1. A process for creating a bioadhesive liposome formed by covalently bonding a recognizing substance to a liposome comprising the steps of:

(a) providing a reaction vessel containing a liposome having phosphatidylethanolamine;

(b) admixing a recognizing substance of collagen to the reaction vessel to form a reaction mixture with the liposome;

(c) buffering the reaction mixture;

(d) admixing a crosslinking reagent to the buffered reaction mixture;

(e) incubating the buffered reaction mixture and reagent for a period of time sufficient to form the covalent bonding of amine residues of the collagen recognizing substance and liposome to create a bioadhesive liposome; and

,

(f) separating and removing excess unreacted and noncovalently bound materials from the bioadhesive liposome.

2. The process of claim 1 wherein the liposome is selected from the group consisting of multilamellar vesicles and unilamellar vesicles.

3. The process of claim 2 wherein the unilamellar vesicles include microemulsified liposomes and large unilamellar vesicles.

4. The process of claim 1 wherein the crosslinking reagent is glutaraldehyde.

5. The process of claim 1 wherein the crosslinking reagent is a bifunctional oxirane.

6. The process of claim 1 where in the crosslinking reagent is ethylene glycol diglycidyl ether.

7. A bioadhesive liposome produced according to the method of claim 1.

8. A bioadhesive liposome designed for the sustained release of a therapeutic substance comprising a liposome component, the therapeutic substance encapsulated by the liposome component, and a collagen recognizing substance component bonded to the liposomal surface.

9. The bioadhesive liposome of claim 8 wherein the collogen recognizing substance component confers to the liposome component target specificity for and retention at a designated biological target site for release of the theraputic substance.

10. The bioadhesive liposome of claim 9 wherein the collagen recognizing substance component confers to the liposome component target specificity for and retention at the designated biological target site for release of the therapeutic substance from the liposome component without interference from the attachment of the collagen recognizing substance.

11. A modified liposome comprising a liposome having phosphatidylethanolamine as a provider of a primary amine residue covalently linked by a crosslinking reagent to a target recognizing substance which is collagen.

12. The modified liposome of claim 11 wherein the liposome and collagen are covalently linked by glutaraldehyde.