Submicron emulsions as vaccine adjuvants

• US 5,961,970 A
• Filed: 04/29/1996
• Issued: 10/05/1999
• Est. Priority Date: 10/29/1993
• Status: Expired due to Fees

• Current Assignee: Pharmos Corporation, United States Department Of Health And Human Services
• Original Assignee: Pharmos Corporation, The United States Of America As Represented By The Secretary Of The Army
• Inventors: Friedman, Doron, Lowell, George H., Amselem, Shimon, Aviv, Haim
• Primary Examiner(s): MINNIFIELD, NITA M
• Application Number:
  US08/637,756
• Time in Patent Office: 1,254 Days
• Field of Search: 424/279.1, 424/278.1, 424/283.1, 424/455, 424/452, 424/490, 424/450, 424/462, 424/184.1, 424/93.1
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 2039/541 Mucosal route
• CPC Class Codes: A61K 2039/543 intranasal
• 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 2039/55544 Bacterial toxins
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 2039/6037 Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
• CPC Class Codes: A61K 2039/6068 Other bacterial proteins, e.g. OMP
• CPC Class Codes: A61K 39/008 Leishmania antigens
• CPC Class Codes: A61K 39/085 Staphylococcus
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/21 Retroviridae, e.g. equine infectious anemia virus
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12N 2740/16134 : 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
• CPC Class Codes: Y10S 977/906 : Drug delivery
• CPC Class Codes: Y10S 977/907 : Liposome
• CPC Class Codes: Y10S 977/915 : Therapeutic or pharmaceutical composition
• CPC Class Codes: Y10S 977/917 : Vaccine
• CPC Class Codes: Y10S 977/918 : Immunological

Abstract

A vaccine adjuvant composition of an oil-in-water submicron emulsion that has about 0.5 to 50% of a first component of an oil, about 0.1 to 10% of a second component of an emulsifier, about 0.05 to 5% of a nonionic surfactant, about 0.00001 to 1% of an immunogen, and an aqueous continuous phase. This submicron emulsion has a mean droplet size in the range of between about 0.03 and 0.5 μm, and preferably 0.05 and 0.2 μm.

Claims

1. A vaccine adjuvant composition of an oil-in-water submicron emulsion consisting essentially of about 0.5 to 50% of a first component of an oil, about 0.1 to 10% of a second component of an emulsifier, wherein the emulsifier is a phospholipid compound or a mixture of phospholipids selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, phosphatidic acid, sphingomyelin and cardiolipin, about 0.05 to 5% of a non-ionic surfactant, about 0.00001, to 1% of an immunogen, and an aqueous continuous phase, said submicron emulsion having a mean droplet size in the range of between about 0.03 and 0.5 μ

m, which composition is substantially free of added amounts of muramyl peptides or their lipophilic derivatives.

2. The emulsion of claim 1 wherein said mean droplet size is between about 0.05 and 0.2 μ

m.

3. The emulsion of claim 1, wherein said first component is selected from the group consisting of a medium chain triglyceride oil, a vegetable oil and mixtures thereof.

4. The emulsion of claim 3 wherein said first component is present in an amount of about 1 to 20%.

5. The emulsion of claim 1 wherein said emulsifier is present in an amount of about 0.2 to 5%.

6. The emulsion of claim 1 wherein said surfactant is selected from the group consisting of fatty alcohols, polyethylene glycol esters of fatty acids, polyethoxylated fatty acid esters, polyethoxylated fatty alcohol ethers and polyethoxylated alkylphenyl ethers.

7. The emulsion of claim 1 wherein said surfactant is an alkylene oxide condensate of an organic compound having one or more hydroxyl groups.

8. The emulsion of claim 7 wherein said non-ionic surfactant is present in an amount of about 0.1 to 5%.

9. The emulsion of claim 1 wherein said first component is present in an amount of about 1 to 20%, and the second component and the non-ionic surfactant are each present in an amount of about 0.1 to 2%.

10. The composition of claim 1 wherein said immunogen is selected from the group consisting of hydrophilic, lipophilic and amphiphilic immunogens.

11. The composition of claim 1 wherein said immunogen is selected from the group consisting of native, recombinant and synthetic peptides, proteins and glycoproteins derived from a source selected from the group consisting of bacteria, viruses and parasites.

12. The composition of claim 11 wherein said immunogen is the gp160 envelope protein of the HIV virus, or a fragment thereof.

13. The composition of claim 11 wherein said immunogen is the surface glycoprotein of a Leishmania parasite or a fragment thereof.

14. The composition of claim 13 wherein said surface glycoprotein or peptide is covalently conjugated to a hydrophobic foot.

15. The composition of claim 14 wherein said hydrophobic foot is lauryl-cysteine.

16. The composition of claim 1 wherein said immunogen is a protein toxoid.

17. The composition of claim 16 wherein said immunogen is Staphylococcus Enterotoxin B toxoid.

18. The composition of claim 1 wherein said immunogen is selected from the group consisting of an oligosaccharide, a polysaccharide and a lipopolysaccharide.

19. The composition of claim 18 wherein said immunogen is a lipopolysaccharide from Shigella flexneri.

20. The composition of claim 1 wherein said immunogen is completed to proteosomes.

21. A method for preparing the composition of claim 1 by an intrinsic procedure, which comprises adding the immunogen to the oil phase before emulsification of the water and oil phases.

22. A method for preparing the composition of claim 1 by an intrinsic procedure, which comprises adding the immunogen to the water phase before emulsification of water and oil phases.

23. A method for preparing the composition of claim 1 by an extrinsic procedure, which comprises preparing a submicron emulsion from the oil, surfactant and emulsifier components, and adding the immunogen externally by mixing it with the previously prepared submicron emulsion.

24. A method for providing enhanced immunogenicity in a subject which comprises administering the composition of claim 1, to the subject to introduce the immunogen therein.

25. The method of claim 24 which further comprises administering the composition parenterally, orally, intranasally or topically to the subject.