Glycolipopeptide and uses thereof

• US 20090041836A1
• Filed: 07/03/2008
• Published: 02/12/2009
• Est. Priority Date: 01/03/2006
• Status: Active Grant

• Current Assignee: The University of Georgia Research Foundation, Inc.
• Original Assignee: The University of Georgia Research Foundation, Inc.
• Inventors: Wolfert, Margaretha A., Ingale, Sampat, Boons, Geert-Jan, Buskas, Therese
• Granted Patent: US 7,820,797 B2
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55577 Saponins; Quil A; QS21; ISCOMS
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 2039/6018 Lipids, e.g. in lipopeptides
• CPC Class Codes: A61K 2039/6087 Polysaccharides; Lipopolysaccharides [LPS]
• 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 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/02 Bacterial antigens
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/385 Haptens or antigens, bound to carriers
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: B01J 13/02 Making microcapsules or microballoons for medical preparations A61K9/50
• CPC Class Codes: B01J 13/04 by physical processes, e.g. drying, spraying
• CPC Class Codes: C07K 14/22 from Neisseriaceae (F)
• CPC Class Codes: C07K 14/4727 Mucins, e.g. human intestinal mucin
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/3076 against structure-related tumour-associated moieties
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 9/00 : Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
• CPC Class Codes: C12N 2770/24234 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: G01N 33/5308 : for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/92 : involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• 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

A glycolipopeptide comprising a carbohydrate component, a peptide component and a lipid component, for use as a therapeutic or prophylactic vaccine. Also provided are monoclonal and polyclonal antibodies that recognize the glycolipopeptide of the invention, as well as uses thereof.

Claims

1. A glycolipopeptide comprising:

at least one carbohydrate component comprising a B-epitope;

at least one peptide component comprising a T-epitope; and

at least one lipid component;

wherein the carbohydrate component and the peptide component are heterologous with respect to each other.

2. The glycolipopeptide of claim 1 wherein the B-epitope is from a microorganism.

3. The glycolipopeptide of claim 2 wherein the microorganism is selected from the group consisting of a virus, a bacterium, a fungus, and a protozoan.

4. The glycolipopeptide of claim 2 wherein the microorganism is a human immunodeficiency virus.

5. The glycolipopeptide of claim 2 wherein the microorganism is a hepatitis C virus.

6. The glycolipopeptide of claim 1 wherein the B epitope is overexpressed on a cancer cell.

7. The glycolipopeptide of claim 1 wherein the carbohydrate component comprises a self-antigen.

8. The glycolipopeptide of claim 7 wherein the self-antigen comprises a MUC-1 glycopeptide.

9. The glycolipopeptide of claim 1 wherein the carbohydrate component comprises a viral antigen.

10. The glycolipopeptide of claim 9 wherein the viral antigen is from human immunodeficiency virus.

11. The glycolipopeptide of claim 9 wherein the viral antigen is from hepatitis C virus.

12. The glycolipopeptide of claim 1 wherein the carbohydrate component comprises a glycoconjugate selected from the group consisting of a glycosylated protein, a glycosylated peptide, a glycosylated lipid, a glycosylated amino acid, a DNA and an RNA.

13. The glycolipopeptide of claim 12 wherein the glycosylated peptide or glycosylated protein comprises a β

-N-acetylglucosamine (β

-O-GlcNAc) modified peptide.

14. The glycolipopeptide of claim 1 comprising a glycosylated peptide, wherein the glycosylated peptide comprises the B-epitope and the T-epitope.

15. The glycolipopeptide of claim 1 wherein the carbohydrate component comprises a heparin sulfate fragment.

16. The glycolipopeptide of claim 1 wherein the lipid component comprises a Toll-like receptor (TLR) ligand.

17. The glycolipopeptide of claim 16 wherein the Toll-like receptor ligand comprises a lipid selected form the group consisting of Pam₂Cys, Pam₃Cys, Pam₂CysSK_n, and Pam₃CysSK_n, wherein n=0, 1, 2, 3, 4 or 5.

18. The glycolipopeptide of claim 1 wherein the lipid component facilitates internalization of the glycolipopeptide by a target cell.

19. The glycolipopeptide of claim 1 wherein the lipid component comprises a component selected from the group consisting of the lipid components listed in Scheme 8.

20. The glycolipopeptide of claim 1 wherein the T-epitope comprises a helper T-epitope.

21. The glycolipopeptide of claim 1 further comprising at least one linker component.

26. A pharmaceutical composition comprising:

the glycolipopeptide of claim 1; and

a pharmaceutically acceptable carrier.

27. The pharmaceutical composition of claim 26 further comprising a liposome.

28. The pharmaceutical composition of claim 27 wherein the glycolipopeptide is covalently or noncovalently incorporated into the liposome.

29. The pharmaceutical composition of claim 28 wherein a plurality of glycolipopeptides are covalently or noncovalently incorporated in the liposome.

30. The pharmaceutical composition of claim 29 wherein at least some of the glycolipopeptides contain different B-epitopes.

31. The pharmaceutical composition of claim 29 wherein at least some of the glycolipopeptides contain the same B-epitopes.

32. The pharmaceutical composition of claim 26 which does not contain an external adjuvant.

33. The pharmaceutical composition of claim 26 further comprising an external adjuvant.

34. The pharmaceutical composition of claim 33 wherein the external adjuvant comprises QS-21.

35. The pharmaceutical composition of claim 26 formulated for use as a vaccine.

36. A method for treating or preventing an infection, disease or disorder in a subject comprising administering to the subject the pharmaceutical composition of claim 26.

37. The method of claim 36 wherein the pharmaceutical composition includes an adjuvant comprising QS-21, and wherein the inclusion of QS-21 skews the immune response of the subject toward a Th1 response, compared to a comparable pharmaceutical composition that does not include QS-21.

38. The method of claim 36 wherein the infection, disease or disorder is caused by a microorganism.

39. The method of claim 38 wherein the microorganism is selected from the group consisting of a virus, a bacterium, a fungus, and a protozoan.

40. The method of claim 36 wherein the infection, disease or disorder is caused by a human immunodeficiency virus.

41. The method of claim 36 wherein the infection, disease or disorder is caused by a hepatitis C virus.

42. The method of claim 36 wherein the disease is cancer, a precancerous condition, or an autoimmune disease.

43. A polyclonal or monoclonal antibody that binds to the glycolipopeptide of claim 1.

44. A polyclonal or monoclonal antibody that binds to the glycolipopeptide of claim 8.

45. A polyclonal or monoclonal antibody that binds to the glycolipopeptide of claim 13.

46. A polyclonal or monoclonal antibody that binds to the glycolipopeptide of claim 15.

47. A hybridoma that produces a monoclonal antibody that binds to the glycolipopeptide of claim 1.

49. A pharmaceutical composition comprising:

a monoclonal or polyclonal antibody that binds to the glycolipopeptide of claim 1; and

a pharmaceutically acceptable carrier.

54. A diagnostic kit comprising:

a monoclonal antibody that binds to the glycolipopeptide of claim 1;

packaging; and

instructions for use.

55. The diagnostic kit of claim 54 wherein the monoclonal antibody is conjugated to a detectable label.

56. The diagnostic kit of claim 54 further comprising a second antibody that binds to the monoclonal antibody.

57. The diagnostic kit of claim 56 wherein the second antibody is conjugated to a detectable label.

58. A method for detecting or diagnosing an infection, disease or disorder in a subject, the method comprising:

incubating a biological sample with an antibody that binds to the glycolipopeptide of claim 1; and

detecting binding of the antibody to a component in the biological sample;

wherein binding of the antibody to a sample component is indicative of the presence of the infection, disease or disorder in the subject.

59. The method of claim 58 further comprising:

quantitating the level of antibody binding to the sample component;

quantitating the level of antibody binding to components in a comparable non-diseased sample; and

comparing the binding levels;

wherein a change in antibody binding in the biological sample compared to the non-diseased sample is indicative of the presence of the infection, disease or disorder in the subject.

22. A glycolipopeptide comprising:

at least one carbohydrate component comprising a self-antigen comprising a B-epitope;

at least one peptide component comprising a T-epitope; and

at least one lipid component.

23. The glycolipopeptide of claim 22 wherein the lipid component comprises a Toll-like receptor ligand.

24. The glycolipopeptide of claim 22 wherein the self-antigen comprises a MUC-1 glycopeptide.

25. The glycolipopeptide of claim 22 wherein the self-antigen comprises a β

-N-acetylglucosamine (β

-O-GlcNAc) modified peptide.

48. A hybridoma cell line selected from the group consisting of hybridoma 1F5.D6, hybridoma 9D1.E4, hybridoma 5H11.H6, and hybridoma 18B10.C7.

50. A method for making a glycolipopeptide comprising:

contacting a candidate compound with a target cell containing a Toll-like receptor (TLR);

determining whether the candidate compound binds to the TLR so as to identify a TLR ligand; and

covalently linking the identified TLR ligand to a carbohydrate component comprising a B-epitope and a peptide component comprising a T-epitope to yield a glycolipopeptide comprising the carbohydrate component, the peptide component, and the TLR ligand.

51. The method of claim 50 further comprising determining whether the candidate compound is internalized by the target cell.

52. The method of claim 50 wherein the candidate compound comprises a lipid.

53. A glycolipopeptide made according to the method of claim 50.