Molecules, compositions, methods and kits for applications associated with flaviviruses

• US 20040242483A1
• Filed: 01/29/2004
• Published: 12/02/2004
• Est. Priority Date: 01/22/2003
• Status: Active Grant

• Current Assignee: National University of Singapore
• Original Assignee: National University of Singapore
• Inventors: Hann, Justin Chu Jang, Lee, Mary Ng Mah
• Granted Patent: US 7,449,321 B2
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 31/737 Sulfated polysaccharides, e.g. chondroitin sulfate, dermatan sulfate A61K31/727 takes precedence
• CPC Class Codes: A61K 38/162 from virus
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/70557 Integrin beta3-subunit-containing molecules, e.g. CD41, CD51, CD61
• CPC Class Codes: C07K 16/1081 Togaviridae, e.g. flavivirus, rubella virus, hog cholera virus
• CPC Class Codes: C12N 2770/24122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: G01N 33/56983 Viruses
• 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 method for controlling a flavivirus entry into a cell, kits for assaying the flavivirus entry into the cell, and methods of treating and preventing flaviviruses infections are disclosed, together with vaccine and pharmaceutical compositions.

Claims

12. A method for controlling flavivirus entry into a cell, comprising administering to the cell an agent interfering with the expression of a flavivirus receptor protein, the receptor protein being integrin.

13. The method of claim 12, wherein the agent is a siRNA against the integrin.

17. A kit for controlling entry of a flavivirus into a cell, the kit comprising:

the flavivirus; and

an agent interfering with expression of an integrin, the first flavivirus and the interfering agent to be used according to the method of claim 12.

18. The kit of claim 17, wherein the interfering agent interfering with the expression of an integrin is an SiRNA against the integrin.

19. A method for controlling a flavivirus entry into a cell, the cell having a plasma memebra, the method comprising administering to the cell an interfering agent functionally interfering with an ATPase in the plasma membrane of the cell.

1. A method for controlling a flavivirus entry into a cell, comprising administering to the cell an agent functionally interfering with a flavivirus receptor protein, the flavivirus receptor protein being an integrin.

2. The method of claim 1, wherein the integrin comprises integrin subunit β

3 or integrin subunit α

V.

3. The method of claim 1, wherein the integrin is an α

Vβ

3 integrin.

4. The method of claim 3, wherein the flavivirus receptor protein is polypeptide from Vero Cells, having approximately a 105 KDa molecular weight and comprising portions with a sequence substantially homologous to SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4 and SEQ ID NO;

5.

5. The method of claim 1, wherein the agent functionally interfering with a flavivirus receptor protein is a functional blocking antibody against the integrin.

6. The method of claim 1, wherein the agent functionally interfering with a flavivirus receptor protein is a competitive ligand for the integrin.

7. The method of claim 6, wherein the competitive ligand is an RGD peptide.

8. The method of claim 6, wherein the competitive ligand is a natural ligand selected from the group consisting of fibronectin, vitronectin, laminin and chrondriotin.

9. The method of claim 6, wherein the flavivirus is a member of Japanese encephalitis serocomplex.

10. The method of claim 9, wherein the flavivirus is West Nile Virus.

11. The method of claim 1, wherein the cell is a cell of a vertebrate.

14. A kit for controlling entry of a flavivirus into a cell, the kit comprising:

the flavivirus; and

an agent functionally interfering with an integrin, the flavivirus and the interfering agent to be used according to the method of claim 1.

15. The kit of claim 14, wherein the agent functionally interfering with an integrin is a functional blocking antibody against the integrin.

16. The kit of claim 14, wherein the agent functionally interfering with an integrin is a competitive ligand for the integrin.

20. The method of claim 19, wherein the interfering agent is a functional blocking antibody against the ATPase.

21. The method of claim 19, wherein the agent functionally interfering with a flavivirus receptor protein is a competitive ligand for the ATPase.

22. The kit of claim 21, wherein the interfering agent is a functional blocking antibody against the ATPase.

21-1. A kit for controlling entry of a flavivirus into a cell, the cell having a plasma membrane, the kit comprising:

the flavivirus; and

an agent functionally interfering with an ATPase located in the plasma membrane of the cell, the flavivirus and the intefering agent to be used according to the method of claim 19.

23. A method for controlling a flavivirus entry into a cell, comprising administering to the cell an agent functionally interfering with a flavivirus receptor protein, the receptor protein being a neurotensin receptor.

24. The method of claim 23, wherein the agent functionally interfering with a flavivirus receptor protein is a functional blocking antibody against the neurotensin receptor.

25. The method of claim 23, wherein the agent functionally interfering with a flavivirus receptor protein is a competitive ligand for the neurotensin receptor.

26. The method of claim 25, wherein the competitive ligand is neurotensin.

27. A kit for controlling entry of a flavivirus into a cell, the kit comprising:

the flavivirus; and

an agent functionally interfering with a neurotensin receptor located in the plasma membrane of the cell, the flavivirus and the intefering agent to be used according to the method of claim 23.

28. A method for controlling entry of a flavivirus into a cell, the flavivirus exhibiting a flavivirus envelope protein, the flavivirus envelope protein comprising a domain III of the flavivirus envelope protein, the method comprising administering to the cell an agent functionally interfering with the domain III of the flavivirus envelope protein.

29. The method of claim 28, wherein the domain III has a sequence substantially homologous to SEQ ID NO:

19 or SEQ ID NO;

21.

30. The method of claim 29, wherein the agent is an antibody against the domain III.

31. A method for treating a flavivirus infection in a vertebrate, the flavivirus exhibiting a flavivirus envelope protein, the flavivirus envelope protein comprising a domain III, the method comprising

administering to the vertebrate a pharmaceutically effective amount of an agent functionally inhibiting the domain III of the envelope protein of the flavivirus.

32. The method of claim 31, wherein the domain has a sequence substantially homologous to SEQ ID NO:

19 or SEQ ID NO;

21.

33. The method of claim 31, wherein the agent is a functional blocking antibody against the domain III.

34. The method of claim 31, wherein the agent is a competitive ligand of domain III.

35. The method of claim 31, wherein the flavivirus is a member of the Japanese encephalitis serocomplex.

36. The method of claim 35, wherein the flavivirus is West Nile Virus.

37. The method of claim 31, wherein the vertebrate is a human being.

38. Pharmaceutical composition for the treatment of a flavivirus infection in a vertebrate, the flavivirus exhibiting an envelope protein comprising a domain III, the pharmaceutical composition comprising:

a pharmaceutically effective amount of an agent functionally inhibiting the domain III of the envelope protein and a pharmaceutically acceptable carrier, vehicle or auxiliary agent.

39. The pharmaceutical composition of claim 38, wherein the interfering agent is a functional blocking antibody against the domain III.

40. The pharmaceutical composition of claim 40, wherein the interfering agent is competitive ligand for the domain III.

41. A method for inducing immunity to a flavivirus in a vertebrate. susceptible to infection of the flavivirus, the flavivirus exhibiting an envelope protein comprising a domain ll, the method comprising:

administering to the vertebrate an immunogenic amount of a polypeptide comprising the domain III of the envelope protein of the flavivirus.

42. The method of claim 41, wherein the domain III comprises a portion having a sequence substantially homologous to SEQ ID NO:

19 or SEQ ID NO;

21.

43. A vaccine for a flavivirus, the flavivirus exhibiting an envelope protein comprising a domain III, the vaccine comprising, as an active agent, a polypeptide comprising the domain III of the envelope protein of the flavivirus.

44. A method for diagnosing a flavivirus infection in a vertebrate susceptible to be infected by the flavivirus, the method comprising:

contacting a sample tissue from the vertebrate, with an integrin or neurotensin protein associated with an identifier;

detecting presence or absence of a flavivirus-integrin complex or flavivirus-neurotensin complex by detecting presence of the identifier.

45. A kit for the diagnosis of flavivirus infection in a vertebrate, susceptible to be infected with the flavivirus, the flavivirus exhibiting an envelope protein comprising a domain III, the kit comprising:

at least one agent able to bind the domain ll, associated with an identifier, and

one or more reagents able to detect the identifier, the at least one agent able to bind domain III and the one or more reagents to be used according to the method of claim 44.

46. A diagnostic method to analyze a cell susceptibility to flavivirus infection, the method comprising

contacting the cell with an identifier for the presence or expression of an integrin, neurotensin receptor and or ATP-ase;

and

detecting the presence of the identifier.

47. A kit to analyze a cell susceptibility to flavivirus infection, the kit comprising:

an identifier for the presence or expression of an integrin, neurotensin receptor and or ATP-ase; and

a reagent able to detect the presence of the identifier, the identifier and the reagent to be used in the method of claim 46.

48. Isolated and purified polypeptide from Vero Cells, having approximately a 105 KDa molecular weight and comprising portions with a sequence substantially homologous to SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4 and SEQ ID NO;

5.

49. A polypeptide having a sequence substantially homologous to SEQ ID NO:

20 or SEQ ID NO;

21.

50. An antibody against a polypeptide from Vero Cells, having approximately a 105 KDa weight and comprising portions with a sequence substantially homologous to SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4 and SEQ ID NO;

5.

51. An antibody against a polypeptide having a sequence substantially homologous to SEQ ID NO:

19 or SEQ ID NO;

21.