Process for microencapsulating cells

• US 6,033,888 A
• Filed: 07/10/1997
• Issued: 03/07/2000
• Est. Priority Date: 05/11/1992
• Status: Expired due to Fees

• Current Assignee: University of Florida (State University System of Florida)
• Original Assignee: University of Florida (State University System of Florida)
• Inventors: Vaghefi, Farid, Batich, Chris
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US08/891,189
• Time in Patent Office: 971 Days
• Field of Search: 264/4.32, 264/4.33, 424/488, 424/497, 424/499, 514/963, 514/965, 514/891, 435/179, 435/178, 427/2.14
• US Class Current: 435/178
• CPC Class Codes: A61K 35/74 Bacteria therapeutic use of a bacterial protein A61K38/00
• CPC Class Codes: A61K 9/1652 Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin homeopathic globules A61K9/1623
• CPC Class Codes: A61K 9/5026 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• CPC Class Codes: A61K 9/5031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5073 having two or more different coatings optionally including drug-containing subcoatings
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres

Abstract

Oxalate-degrading enzymes and bacteria were encapsulated for both enteric and intraperitoneal administration. We have shown that via alginate microencapsulation of Oxalobacter formigenes, enzymatic activity was retained for several months. A new process was developed which strengthened the aliginate microcapsules and their tolerance to citrate treatment. Much smaller (30-50 μm) aliginate microcapsules were made for injection as means of impantation. For oral administration, multi-encapsulated microspheres of cellulose acetate phthalate in poly-2-vinylpyridine (pKa=3.5) were prepared to protect the enzymes from gastric juices.

Claims

1. In a method for preparing microcapsules of a desired material wherein said method comprises the following steps:

(a) mixing said desired material to be encapsulated with a matrix-forming composition;

(b) making droplets of the mixture of part (a) and introducing said droplets into a gelling solution to form droplets;

(c) treating the gel droplets formed in part (b) with a polyelectrolyte to form a microcapsule;

an improvement which comprises the addition of at least one more polyelectrolyte layer and at least one more matrix-forming layer to the microcapsule of part (c), wherein said polyelectrolyte is positively charged and said matrix-forming composition comprises alginate, wherein a matrix-forming layer is the last layer to be added to said microcapsule.

2. The method, according to claim 1, wherein part (a) comprises mixing said material to be encapsulated with a solution of alginate having a concentration of about 0.5% to about 2.5% of alginate.

3. The method, according to claim 2, wherein said alginate has a molecular weight of about 50 kD to about 150 kD.

4. The method, according to claim 1, wherein at least one additional layer of alginate is added to said microcapsule by dipping said microcapsule into an alginate solution wherein said alginate solution has a concentration of approximately 10% of the concentration of the alginate solution used in part (a).

5. The method, according to claim 4, wherein said polyelectrolyte is poly-L-lysine.

6. The method, according to claim 5, which comprises adding three additional layers of alginate and three additional layers of poly-L-lysine with an alginate layer being the last to be added.

7. The method, according to claim 6, which further comprises a liquefaction step.

8. The method, according to claim 7, wherein said liquefaction step comprises treatment with citrate.

9. The method, according to claim 8, wherein said citrate treatment is with a 1.5% citrate solution for at least 5 minutes.

10. The method, according to claim 9, wherein said encapsulated material is selected from the group consisting of bacteria polypeptides and islet cells.

11. The method, according to claim 10, wherein said bacteria are Oxalobacter formigenes.

12. The method, according to claim 1, wherein said gelling solution is a calcium chloride solution.

13. A microcapsule produced by the process of claim 1.