Therapeutic treatment and prevention of infections with a bioactive materials encapsulated within a biodegradable-biocompatible polymeric matrix

• US 6,844,010 B1
• Filed: 07/18/2000
• Issued: 01/18/2005
• Est. Priority Date: 03/16/1984
• Status: Expired due to Fees

• Current Assignee: United States Army (U.S. Department Of Defense)
• Original Assignee: The United States Of America As Represented By The Secretary Of The Army
• Inventors: Brown, William, Tice, Thomas R., Jarboe, Daniel L., Jeyanthi, Ramasubbu, Jacob, Elliot, Reid, Robert H., Thies, Curt, Boedeker, Edgar C., Setterstrom, Jean A., Cassels, Frederick, Van Hamont, John E., McQueen, Charles E., Roberts, F. Donald, Friden, Phil
• Primary Examiner(s): Carlson, Karen Cochrane
• Assistant Examiner(s): Robinson, Hope A.
• Application Number:
  US09/618,577
• Time in Patent Office: 1,645 Days
• Field of Search: 424/501, 424/486, 424/499, 424/493, 424/491, 424/422, 424/426, 514/570, 514/2, 514/12, 530/350, 530/300
• US Class Current: 424/501
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2039/55505 Inorganic adjuvants
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/09 Luteinising hormone-releasing hormone [LHRH] , i.e. Gonadotropin-releasing hormone [GnRH]; Related peptides
• CPC Class Codes: A61K 38/1729 Cationic antimicrobial peptides, e.g. defensins
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/0258 Escherichia
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/292 Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 9/1611 Inorganic compounds
• CPC Class Codes: A61K 9/1617 Organic compounds, e.g. phospholipids, fats
• CPC Class Codes: A61K 9/1635 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• CPC Class Codes: A61K 9/1641 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
• CPC Class Codes: A61K 9/1647 Polyesters, e.g. poly(lactide-co-glycolide)
• 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: C12N 2730/10134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Novel burst-free, sustained release biocompatible and biodegrable microcapsules which can be programmed to release their active core for variable durations ranging from 1-100 days in an aqueous physiological environment. The microcapsules are comprised of a core of polypeptide or other biologically active agent encapsulated in a matrix of poly(lactide/glycolide) copolymer, which may contain a pharmaceutically-acceptable adjuvant, as a blend of upcapped free carboxyl end group, and end-capped forms ranging in ratios from 100/0 to 1/99.

Claims

1. A process for preparing controlled release compositions characterized by burst-free, sustained, programmable release of biologically active agents or active core, comprising:

dissolving biodegradable poly(lactide/glycolide), in uncapped from and biodegradable poly(lactide/glycolide) in end-capped form in methylene chloride, and dissolving a biologically active agent or active core in water;

adding the aqueous layer to the polymer solution and emulsifying to provide an inner water-in-oil (w/o) emulsion;

stabilizing the w/o emulsion in a solvent-saturated aqueous phase containing an oil-in-water (o/w) emulsifier;

adding said w/o emulsion to an external aqueous layer containing oil-in-water emulsifier to form a ternary emulsion; and

stirring the resulting water-in-oil (w/o/w) emulsion for sufficient time to remove said solvent, and rinsing hardened microcapsules with water and lyophilizing said hardened microcapsules.

2. The process of claim 1 wherein a solvent-saturated external aqueous phase is added to emulsify the inner w/o emulsion prior to addition of the external aqueous layer, to provide microcapsules of narrow size distribution range between 0.05-500 μ

m.

3. The process of claim 1 wherein a low temperature of about 0-4 degree C. is provided during preparation of the inner w/o emulsion, and a low temperature of about 4-20 degree C. is provided during preparation of the w/o/w emulsion to provide a stable emulsion and efficient encapsulation.

4. The process of claim 1 wherein a 100/0 blend of uncapped and end-capped polymer is used to provide release of the biologically active agents or active core in a continuous and sustained manner without a lag phase.

5. The process of claim 1, wherein the biologically active agent is a polypeptide.

6. The process of claim 5, wherein said polypeptide is any of the vaccine agents against enterotoxigenic E. coli selected from the group consisting CFA/I, CFA/II, CS1, CS3, CS6, and CS17, ETEC-related enterotoxins, and combinations thereof.

7. The process of claim 6, wherein said polypeptide is CFA/I.

8. The process of claim 7, wherein said CFA/I polypeptide is synthetic and is selected from the group of synthetic peptides containing the CFA/I pilus protein T-cell epitopes:

4(Asn-Ile-Thr-Val-Thr-Ala-Ser-Val-Asp-Pro),

8(Thr-Ala-Ser-Val-Asp-Pro-Val-Ile-Asp-Leu),

12(Asp-Pro-Val-Ile-Asp-Leu-Leu-Gln-Ala-Asp),

15(Ile-Asp-Leu-Leu-Gln-Ala-Asp-Gly-Asn-Ala),

20(Ala-Asp-Gly-Asn-Ala-Leu-Pro-Ser-Ala-Val),

26(Pro-Ser-Ala-Val-Lys-Leu-Ala-Tyr-Ser-Pro),

72(Leu-Asn-Ser-Thr-Val-Gln-Met-Pro-Ile-Ser),

78(Met-Pro-Ile-Ser-Val-Ser-Trp-Gly-Gly-Gln),

87(Gln-Val-Leu-Ser-Thr-Thr-Ala-Lys-Glu-Phe),

126(Ala-Gly-Thr-Ala-Pro-Thr-Ala-Gly-Asn-Tyr), and

133(Gly-Asn-Tyr-Ser-Gly-Val-Val-Ser-Leu-Val), and mixtures thereof;

synthetic peptides containing CFA/I pilus protein B-cell (antibody) epitopes;

3 (Lys-Ana-Ile-Thr-Val-Thr-Ala-Ser-Val),

11 (Val-Asp-Pro-Val-Ile-Asp-Leu-Leu-Gln-Ala-Asp),

22(Gly-Asn-Ala-Leu-Pro-Ser-Ala-Val),

32(Ala-Tyr-Ser-Pro-Ala-Ser-Lys-Thr-Phe-Lys-Thr-Phe-Glu-Ser-Tyr-Arg-Val),

32(Ala-Tyr-Ser-Pro-Ala-Ser-Lys-Thr-Phe),

38(Lys-Thr-Phe-Glu-Ser-Tyr-Arg-Val),

66(Pro-Gln-Leu-Thr-Asp-Val-Leu-Asn-Ser),

93(Ala-Lys-Glu-Phe-Glu-Ala-Ala-Ala),

124(Lys-Thr-Ala-Gly-Thr-Ala-Pro-Thr),

127(Gly-Thr-Ala-Pro-Thr-Ala-Gly-Asn-Tyr-Ser),

124(Lys-Thr-Ala-Gly-Thr-Ala-Pro-Thr-Ala-Gly-Asn-Thr-Ser), and mixtures thereof; and

synthetic peptides containing CFA/I pilus protein T-cell and B-cell (antibody) epitopes;

3(Lys-Asn-Ile-Thr-Val-Thr-Ala-Ser-Val-Asp-Pro),

8(Thr-Ala-Ser-Val-Asp-Pro-Val-Ile-Asp-Leu-Leu-Gln-Ala-Asp),

11(Val-Asp-Pro-Val-Ile-Asp-Leu-Leu-Gln-Ala-Asp),

20(Ala-Asp-Gly-Asn-Ala-Leu-Pro-Ser-Ala-Val),

124(Lys-Thr-Ala-Gly-Thr-Ala-Pro-Thr-Ala-Gly-Asn-Tyr-Ser), and

126(Ala-Gly-Thr-Ala-Pro-Thr-Ala-Gly-Asn-Tyr-Ser), and mixtures thereof.

9. The process of claim 1, wherein release rates and duration are achieved by blending said uncapped and said end-capped forms of poly(lactide/glycolide) polymer in different amounts within the same microcapsule, wherein the greater the amount of uncapped form of poly(lactide/glycolide) the faster the release rate and the shorter the release duration.

10. The process of claim 1, wherein when the ratio of uncapped polymer to end-capped polymer is increased, the release rate of the active ingredient increases.

11. The process of claim 1, wherein the uncapped polymer and end-capped polymer is present in ratios ranging from 100/0 to 1/99, respectively.

12. The process of claim 1, wherein the uncapped polymer and end-capped polymer is present in ratios ranging from 90/10 to 40/60.

13. The process of claim 1, wherein the ratio between the lactide and glycotide glycolide (L/G) component is within the range of 40/60 to 0/100.

14. The process of claim 1, wherein the ratio between the lactide and glycolide (L/G) component is within the range of 90/10 to 40/60.

15. The process of claim 1, wherein the ratio between the lactide and glycolide (L/G) component is within the range of 48/52 to 52/48.

16. A process for preparing controlled release characterized by burst-free, sustained, programmable release of biologically active agents, comprising:

dissolving biodegradable poly(lactide/glycolide) in end-capped form in methylene chloride, and dissolving a biologically active agent or active core in water;

adding the aqueous layer to the polymer solution and emulsifying to provide an inner water-in-oil emulsion;

stabilizing the w/o emulsion in a solvent-saturated aqueous phase containing an oil-in-water emulsifier;

adding said stabilized w/o emulsion to an external aqueous layer containing oil-in-water (w/o/w) emulsion for sufficient time to remove said solvent; and

rinsing hardened microcapsules with water; and

lyophilizing said hardened microcapsules.

17. The process of claim 16 wherein a solvent-saturated external aqueous phase is added to emulsify the inner w/o emulsion prior to addition of the external aqueous layer, to provide microcapsules of narrow size distribution range between 0.05-500 μ

m.

18. The process of claim 16 wherein a low temperature of about 0-4 degree C. is provided during preparation of the inner w/o emulsion, and a low temperature of about 4-20 degree C. is provided during preparation of the w/o/w emulsion to provide a stable emulsion and efficient encapsulation.