Microparticle carriers of maximal uptake capacity by both M cells and non-M cells

• US RE41,157 E1
• Filed: 11/30/1999
• Issued: 03/02/2010
• Est. Priority Date: 03/16/1984
• Status: Expired due to Term

• 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 R., VanHamont, John E., Reid, Robert H., Thies, Curt, Boedeker, Edgar C.
• Primary Examiner(s): Padmanabhan; Sreeni
• Assistant Examiner(s): MCMILLIAN, KARA RENITA
• Application Number:
  US09/451,321
• Time in Patent Office: 3,745 Days
• Field of Search: 424/491, 424/493, 424/486, 424/497, 424/499, 424/501, 514/788.1, 514/965
• US Class Current: 424/491
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: Y10S 514/965 Discrete particles in supporting matrix

Abstract

In a solvent extraction process for preparing microspheres of a biodegradable polymer, the improvement comprising: preparing a homogenized antigen-sucrose matrix and adding a solvent to the sucrose-antigen matrix to form a solution; preparing a solution of a biodegradable polymer by adding a solvent to the polymer; adding the biodegradable polymer solution to the antigen-sucrose solution; adding an oil to the polymer-sucrose-antigen solution to form an emulsion having a controlled viscosity that corresponds to a predetermined average particle size of distributions of microspheres of biodegradable polymers; centrifuging the emulsion of controlled viscosity and removing the supernatant to obtain microspheres of a predetermined range of particle size distributions of from about 0.5 to about 7.0 micrometers.

In immunostimulating composition comprising an encapsulating-microsphere of the biodegradable polymer has an average particle size distribution such that the majority of the microspheres will be taken up by the villous epithelium section of the intestines of a mammalian subject when administered as a vaccine against diseases caused by entero-pathogenic organisms.A solvent extraction process for preparing microspheres of a biodegradable polymer. The process includes preparing a homogenized antigen-sucrose matrix and adding a solvent to the sucrose-antigen matrix to form a solution. Preparing a solution of a biodegradable polymer by adding a solvent to the polymer. Adding the biodegradable polymer solution to the antigen-sucrose solution. Adding an oil to the polymer-sucrose-antigen solution to form an emulsion having a controlled viscosity that corresponds to a predetermined average particle size of distributions of microspheres of biodegradable polymers. Centrifuging the emulsion of controlled viscosity and removing the supernatant to obtain microspheres of a predetermined range of particle size distributions of from about 0.5 to about 70 micrometers.

Claims

1. In a solvent extraction process for preparing microspheres of an antigen containing biodegradable poly(DL-lactide-co-glycolide), the improvement comprising:

preparing a lyophilized antigen-sucrose matrix;

adding acetonitrile solvent to the antigen-sucrose matrix to form a solution;

preparing a solution of a biodegradable poly (DL-lactide-co-glycolide) polymer by adding acetonitrile solvent to the polymer;

adding the biodegradable poly (DL-lactide-co-glycolide) polymer acetonitrile solution to the antigen-sucrose acetonitrile solution;

adding an oil to the poly (DL-lactide-co-glycolide) polymer-sucrose-antigen solution to form an emulsion having a controlled viscosity, that corresponds to a predetermined average particle size of distributions of microspheres of poly (DL-lactide-co-glycolide) biodegradable polymers of from about 0.5 to about 7.0 micrometers;

centrifuging the emulsion of controlled viscosity and removing a supernatant to obtain microspheres of the predetermined range of particle size distributions.

2. The process of claim 1, wherein the oil is selected with a predefined viscosity to form the microspheres.

3. The process of claim 1, wherein a thickening agent is added to the oil to increase its viscosity.

4. The process of claim 1, wherein the oil is prediluted with an extractant solvent.

5. The process of claim 1, wherein the oil is a paraffin oil in which the viscosity is adjusted by preheating to a temperature of desired viscosity.

6. The process of claim 1, wherein relative ratios between the lactide and glycolide is 50:

50.

7. The process of claim 1, wherein the average particle size distribution is from about 1.0 to about 2.0 micrometers.

8. The process of claim 1, wherein the oil is a paraffin oil.

9. The process of claim 8, wherein the viscosity of the paraffin oil is reduced by diluting it with heptane or iso-octane.

10. The process of claim 2, wherein the viscosity of the oil is reduced to produce larger spheres.

11. The process of claim 3, wherein the thickening agent is polybutylene.

12. A method of controlling average particle size of microspheres containing an agent in a solvent extraction process, comprising:

adjusting a viscosity of an oil to a value that corresponds to a predetermined average particle size of microspheres to control said size of said microspheres;

adding said oil to a biodegradable poly(DL-lactide-co-glycolide) polymer-stabilizer agent solution emulsion;

centrifuging the emulsion;

and removing said microspheres of said predetermined average particle size.