Modified viral particles with immunogenic properties and reduced lipid content useful for treating and preventing infectious diseases

• US 20050032222A1
• Filed: 06/21/2004
• Published: 02/10/2005
• Est. Priority Date: 06/29/2000
• Status: Active Grant

• Current Assignee: Eli Lilly and Company, Lipid Sciences, Inc.
• Original Assignee: Lipid Sciences, Inc.
• Inventors: Cham, Bill E., Bellotti, Marc, Maltais, Jo-Ann B.
• Granted Patent: US 7,407,662 B2
• Time in Patent Office: Days
• US Class Current: 435/456
• CPC Class Codes: A61K 2039/5252 inactivated (killed)
• 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 39/12 Viral antigens
• CPC Class Codes: A61K 39/29 Hepatitis virus
• CPC Class Codes: A61K 39/292 Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: A61L 2/0088 Liquid substances
• CPC Class Codes: A61M 1/3486 Biological, chemical treatment, e.g. chemical precipitation; treatment by absorbents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• 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: C12N 2730/10163 by chemical treatment
• CPC Class Codes: C12N 2740/15034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2740/15063 by chemical treatment
• CPC Class Codes: C12N 2770/20034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2770/24063 by chemical treatment
• CPC Class Codes: C12N 2770/24334 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2770/24363 by chemical treatment
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention relates to a method for reducing the occurrence and severity of infectious diseases, especially infectious diseases in which lipid-containing infectious viral organisms are found in biological fluids, such as blood. The present invention employs solvents useful for extracting lipids from the lipid-containing infectious viral organism thereby creating modified viral particles with reduced infectivity and enhanced antigenicity. The present invention provides vaccine compositions, comprising these modified viral particles with reduced infectivity and enhanced antigenicity, optionally combined with a pharmaceutically acceptable carrier or an immunostimulant. The vaccine composition is administered to a patient to provide protection against the lipid-containing infectious viral organism. The vaccine compositions of the present invention include combination vaccines of modified viral particles obtained from one or more strains of a virus and/or one or more types of virus.

Claims

1. A modified viral particle comprising at least a partially delipidated viral particle, wherein the partially delipidated viral particle:

initiates a positive immune response in a patient; and

,

incites protection against an infectious organism.

2. The modified viral particle of claim 1 wherein the modified viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

3. The modified viral particle of claim 1, wherein the modified viral particle has a lower cholesterol content than an unmodified viral particle.

4. The modified viral particle of claim 3, wherein the lower cholesterol content is at least 20% lower than the unmodified viral particle.

5. The modified viral particle of claim 3, wherein the lower cholesterol content is at least 30% lower than the unmodified viral particle.

6. The modified viral particle of claim 3, wherein the lower cholesterol content is at least 40% lower than the unmodified viral particle.

7. The modified viral particle of claim 1, wherein the modified viral particle has a different buoyant density than an unmodified viral particle.

8. The modified viral particle of claim 1, wherein the modified viral particle is an HIV viral particle or SIV viral particle.

9. The modified viral particle of claim 8, wherein the modified HIV viral particle or the modified SIV viral particle has reduced cholesterol content when compared to an unmodified HIV viral particle or an unmodified SIV viral particle.

10. The modified viral particle of claim 8, wherein the modified HIV viral particle has p24 levels within at least 40% of p24 levels in an unmodified HIV viral particle.

11. The modified viral particle of claim 8, wherein the modified HIV viral particle has p24 levels within at least 30% of p24 levels in an unmodified HIV viral particle.

12. The modified viral particle of claim 8, wherein the modified SIV viral particle has p27 levels within at least 40% of p27 levels in an unmodified SIV viral particle.

13. The modified viral particle of claim 8, wherein the modified SIV viral particle has p27 levels within at least 30% of p27 levels in an unmodified SIV viral particle.

14. A modified viral particle comprising at least a partially delipidated viral particle having:

a lower cholesterol content than an unmodified viral particle; and

,

a different buoyant density than an unmodified viral particle.

15. The modified viral particle of claim 14 wherein the modified viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

16. The modified viral particle of claim 14 wherein the modified viral particle is HIV and has p24 levels within at least 40% of p24 levels in an unmodified HIV viral particle.

17. The modified viral particle of claim 14 wherein the modified viral particle is SIV and has p27 levels within at least 40% of p27 levels in an unmodified SIV viral particle.

18. The modified viral particle of claim 14, wherein the lower cholesterol content is at least 20% lower than the unmodified viral particle.

19. The modified viral particle of claim 14, wherein the lower cholesterol content is at least 30% lower than the unmodified viral particle.

20. The modified viral particle of claim 14, wherein the lower cholesterol content is at least 40% lower than the unmodified viral particle.

21. The modified viral particle of claim 14, the modified viral particle comprises a protein and the protein is gp41, gp120, p24 or p27 protein.

22. The modified viral particle of claim 14, wherein the modified viral particle is HIV, SIV, SARS, hepatitis B, hepatitis C or BVDV.

23. A modified viral particle comprising at least a partially delipidated viral particle, wherein the partially delipidated viral particle is produced by exposing a non-delipidated viral particle to a delipidation process and wherein the partially delipidated viral particle comprises at least one exposed viral antigen that was not exposed in the non-delipidated viral particle.

24. A method for creating a modified viral particle comprising the steps of:

receiving a plurality of viral particles, each having a viral envelope containing lipid, in a fluid;

exposing the viral particles to a delipidation process; and

,

partially delipidating the viral particles wherein the delipidation process at least partially reduces the lipid content of the viral envelope to create the modified viral particle and wherein the modified viral particle is capable of provoking an immune response in a patient.

25. The modified viral particle of claim 24 wherein the viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

26. A method for creating an antigen delivery vehicle comprising the steps of:

receiving a plurality of viral particles, each having a viral envelope containing lipid, in a fluid;

exposing the viral particles to a delipidation process; and

,

partially delipidating the viral particles to create modified viral particles that act as antigen delivery vehicles, wherein the delipidation process at least partially decreases the lipid content of the viral envelope to expose at least one antigen and wherein the at least one antigen is capable of provoking an immune response in a patient.

27. The antigen delivery vehicle of claim 26 wherein the modified viral particle comprises patient specific antigens.

28. The antigen delivery vehicle of claim 26 wherein the viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

29. A vaccine composition, comprising at least a partially delipidated viral particle having at least one exposed viral antigen that was not exposed in the non-delipidated viral particle wherein the partially delipidated viral particle is capable of provoking an immune response when administered to a patient.

30. The vaccine composition of claim 29, wherein the immune response comprises enhanced interferon gamma production by T cells

31. The vaccine composition of claim 30, wherein the T-cells are CD4+or CD8+T-cells.

32. The vaccine composition of claim 29, wherein the immune response comprises proliferation of cells of the immune system.

33. The vaccine composition of claim 29, wherein the partially delipidated viral particle has a lower cholesterol content than the non-delipidated viral particle.

34. The vaccine composition of claim 33, wherein the lower cholesterol content is at least 20% lower than the non-delipidated viral particle.

35. The vaccine composition of claim 33, wherein the lower cholesterol content is at least 30% lower than the non-delipidated viral particle.

36. The vaccine composition of claim 29, comprising partially delipidated viral particles from one or more strains of virus or one or more types of virus.

37. The vaccine composition of claim 29, wherein the partially delipidated viral particle has a different buoyant density than the non-delipidated viral particle.

38. The vaccine composition of claim 29, wherein the partially delipidated viral particle is an HIV viral particle or an SIV viral particle.

39. The vaccine composition of claim 38, wherein the partially delipidated HIV viral particle or the partially delipidated SIV viral particle has reduced cholesterol content when compared to an unmodified HIV viral particle or an unmodified SIV viral particle.

40. The vaccine composition of claim 38, wherein the partially delipidated HIV viral particle has p24 levels within at least 40% of p24 levels in an unmodified HIV viral particle.

41. The vaccine composition of claim 38, wherein the partially delipidated SIV viral particle has p27 levels within at least 40% of p27 levels in an unmodified SIV viral particle.

42. The vaccine composition of claim 29, wherein the partially delipidated viral particle is immunodeficiency virus and the immunodeficiency virus is HIV or SIV.

43. The vaccine composition of claim 29, wherein the partially delipidated viral particle is pestivirus and the pestivirus is BVDV.

44. The vaccine composition of claim 29, wherein the partially delipidated viral particle is hepatitis and the hepatitis is hepatitis B or hepatitis C.

45. The vaccine composition of claim 29, wherein the partially delipidated viral particle is coronavirus and the coronavirus is SARS.

46. The vaccine composition of claim 29, wherein the partially delipidated viral particle comprises a protein and the protein is gp41, gp120, p24 or p27 protein.

47. The vaccine composition of claim 29, wherein the partially delipidated viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

48. A method of providing protection in a patient against an infectious viral particle comprising the step of:

administering to the patient an effective amount of the vaccine composition of claim 29, wherein the amount is effective to provide a protective effect against infection by the infectious viral particle in the patient.

49. The method of claim 48, wherein the infectious viral particle is immunodeficiency virus, hepatitis, pestivirus or coronavirus.

58. The method of claim 48 further comprising administering a pharmaceutically acceptable carrier or an immunostimulant.

50. A method for provoking a positive immune response in a patient having a plurality of lipid-containing viral particles comprising the steps of:

obtaining a fluid containing the lipid-containing viral particles from the patient;

contacting the fluid containing the lipid-containing viral particles with a first organic solvent capable of extracting lipid from the lipid-containing viral particles;

mixing the fluid and the first organic solvent;

permitting organic and aqueous phases to separate;

collecting the aqueous phase containing modified viral particles with reduced lipid content; and

introducing the aqueous phase containing the modified viral particles with reduced lipid content into the patient wherein the modified viral particles with reduced lipid content provoke an immune response in the patient.

51. The method of claim 50, further comprising:

contacting the aqueous phase with charcoal capable of removing the first organic solvent from the aqueous phase.

52. The method of claim 50, wherein after the aqueous phase is collected, the aqueous phase is contacted with charcoal capable of removing the first organic solvent from the aqueous phase, and the aqueous phase containing reduced levels of the first organic solvent is eluted from the charcoal before introducing the aqueous phase containing the modified viral particles with reduced lipid content into the patient.

59. The method of claim 50, wherein the first organic solvent is an alcohol, an ether, an amine, a hydrocarbon, an ester, a surfactant or a combination thereof.

62. The method of claim 50, wherein the first organic solvent is an alcohol, an ether, or a combination thereof.

65. The method of claim 62, wherein the ether is C4 to C8 ether and the alcohol is a C1 to C8 alcohol.

68. The method of claim 50, wherein the fluid is plasma, serum, peritoneal fluid, lymphatic fluid, pleural fluid, pericardial fluid, cerebrospinal fluid, or a fluid of the reproductive system.

53. A method for treating a viral infection in a patient comprising:

removing blood containing a plurality of lipid-containing infectious viral particles from the patient;

obtaining plasma from the blood, the plasma containing the lipid-containing infectious viral particles;

contacting the plasma containing the lipid-containing infectious viral particles with a first organic solvent capable of extracting lipid from the lipid-containing infectious viral particles to produce modified viral particles having reduced lipid content;

mixing the plasma and the first organic solvent;

permitting organic and aqueous phases to separate;

collecting the aqueous phase containing the modified viral particles; and

introducing the aqueous phase containing the modified viral particles into the patient wherein the modified viral particles have at least one exposed viral antigen that was not exposed in the plurality of lipid-containing infectious viral particles.

54. The method of claim 53, wherein after the aqueous phase is collected, the aqueous phase is contacted with charcoal capable of removing the first organic solvent from the aqueous phase and the aqueous phase with reduced levels of first organic solvent is eluted from the charcoal before introducing the aqueous phase containing the modified viral particles into the patient.

55. The method of claim 53, wherein the lipid containing infectious particles are immunodeficiency virus, hepatitis, pestivirus or coronavirus.

60. The method of claim 53, wherein the first organic solvent is an alcohol, an ether, an amine, a hydrocarbon, an ester, a surfactant or a combination thereof.

63. The method of claim 53, wherein the first organic solvent is an alcohol, an ether, or a combination thereof.

66. The method of claim 63 wherein the ether is C4 to C8 ether and the alcohol is a C1 to C8 alcohol.

56. A method for making a vaccine comprising:

contacting a lipid-containing viral particle in a fluid with a first organic solvent capable of extracting lipid from the lipid-containing viral particle;

mixing the fluid and the first organic solvent for a time sufficient to extract lipid from the lipid-containing viral particle;

permitting organic and aqueous phases to separate; and

,

collecting the aqueous phase containing a modified viral particle with reduced lipid content wherein the modified viral particle is capable of provoking an immune response when administered to a patient.

57. The method of claim 56, further comprising:

contacting the aqueous phase with charcoal capable of removing the first organic solvent.

61. The method of claim 56, wherein the first organic solvent is an alcohol, an ether, an amine, a hydrocarbon, an ester, a surfactant or a combination thereof.

64. The method of claim 56, wherein the first organic solvent is an alcohol, an ether, or a combination thereof.

67. The method of claim 64 wherein the ether is C4 to C8 ether and the alcohol is a C1 to C8 alcohol.

69. The method of claim 56, wherein the fluid is plasma, serum, peritoneal fluid, lymphatic fluid, pleural fluid, pericardial fluid, cerebrospinal fluid, or a fluid of the reproductive system.