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Last Updated: March 28, 2024

Claims for Patent: 6,022,864


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Summary for Patent: 6,022,864
Title: Nucleic acid respiratory syncytial virus vaccines
Abstract:Vectors containing a nucleotide sequence coding for an F protein of respiratory syncytial virus (RSV) and a promoter for such sequence, preferably a cytomegalovirus promoter, are described. Such vectors also may contain a further nucleotide sequence located adjacent to the RSV F protein enclosing sequence to enhance the immunoprotective ability of the RSV F protein when expressed in vivo. Such vectors may be used to immunize a host, including a human host, by administration thereto. Such vectors also may be used to produce antibodies for detection of RSV infection in a sample.
Inventor(s): Li; Xiaomao (Thornhill, CA), Ewasyshyn; Mary E. (Willowdale, CA), Sambhara; Suryaprakash (Markham, CA), Klein; Michel H. (Willowdale, CA)
Assignee: Connaught Laboratories Limited (North York, CA)
Application Number:08/973,720
Patent Claims:1. A method of immunizing a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises administering to said host an effective amount of a plasmid vector comprising:

a first nucleotide sequence encoding an RSV F protein that generates antibodies, said first nucleotide sequence being selected from the group consisting of:

a nucleotide sequence having SEQ ID NO:1,

a nucleotide sequence having SEQ ID NO:3,

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:2, and

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:4,

a promoter sequence operatively coupled to said first nucleotide sequence for expression of said RSV F protein in said host, and

a second nucleotide sequence located between said first nucleotide sequence and said promoter sequence and comprising a pair of splice sites to prevent aberrant mRNA splicing and to increase expression of said RSV F protein when expressed in vivo from said vector in said host.

2. The method of claim 1 wherein said host is a human.

3. The method of claim 1 wherein said first nucleotide sequence encodes a full-length RSV F protein having the sequence of SEQ ID NO:2.

4. The method of claim 1 wherein said first nucleotide sequence encodes an RSV F protein from which the transmembrane region is absent and having the sequence of SEQ ID NO:4.

5. The method of claim 1 wherein said promoter sequence is an immediate early cytomegalovirus promoter.

6. The method of claim 1 wherein said second nucleotide sequence is that of rabbit .beta.-globin intron II.

7. The method of claim 2 wherein said promoter sequence is an immediate early cytomegalovirus promoter.

8. A method of using a gene encoding an RSV F protein that generates antibodies to produce an immune response in a host, which comprises:

isolating said gene, said gene having a nucleotide sequence being selected from the group consisting of:

a nucleotide sequence having SEQ ID NO:1,

a nucleotide sequence having SEQ ID NO:3,

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:2, and

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:4;

operatively linking said gene to a promoter sequence to produce a plasmid vector, said promoter sequence directing expression of said RSV F protein when said vector is introduced into a host to produce an immune response to said RSV F protein;

operatively linking said gene in said plasmid vector to a nucleotide sequence comprising a pair of splice sites to prevent aberrant mRNA splicing and to increase expression of said RSV F protein in said host, and

introducing said non-replicating vector into the host.

9. The method of claim 8 wherein said gene encoding an RSV F protein encodes an RSV F protein lacking the transmembrane region.

10. The method of claim 8 wherein said nucleotide sequence is introduced into said vector between said promoter sequence and said gene.

11. The method of claim 9 wherein said promoter sequence comprises the immediate early cytomegalovirus promoter.

12. The method of claim 10 wherein said nucleotide sequence is that of rabbit .beta.-globin intron II.

13. A method of producing a vaccine for protection of a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises:

isolating a first nucleotide sequence encoding an RSV F protein that generates antibodies, said first nucleotide sequence being selected from the group consisting of:

a nucleotide sequence having SEQ ID NO:1,

a nucleotide sequence having SEQ ID NO.3,

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:2, and

a nucleotide sequence encoding an amino acid sequence having SEQ ID NO:4;

operatively linking said first nucleotide sequence to a promoter sequence to produce a plasmid vector, the promoter sequence directing expression of said RSV F protein when introduced into a host to produce an immune response to said RSV F protein;

operatively linking said first nucleotide sequence to a second nucleotide sequence between said first nucleotide sequence and said promoter sequence in said vector, said second nucleotide sequence comprising a pair of splice sites to prevent aberrant mRNA splicing and to increase expression of said RSV F protein when expressed in vivo from the vector in a host, and

formulating said plasmid vector as a vaccine for in vivo administration.

14. A vaccine produced by the method of claim 13.

15. The method of claim 13 wherein said vector is selected from group consisting of pXL1 and pXL2.

16. The method of claim 13 wherein said first nucleotide sequence encodes a full-length RSV F protein having an amino acid sequence having SEQ ID NO:2.

17. The method of claim 13 wherein said first nucleotide sequence encodes a RSV F protein from which the transmembrane region is absent and having an amino acid sequence having SEQ ID NO:4.

18. A method of producing a vaccine for protection of a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises:

isolating a nucleotide sequence encoding an RSV F protein from which the transmembrane region is absent, said nucleotide sequence having SEQ ID NO:3 or encoding an RSV F protein having the amino acid sequence having SEQ ID NO:4;

operatively linking said nucleotide sequence to a promoter sequence to produce a plasmid vector, the promoter sequence directing expression of said RSV F protein when introduced into a host to produce an immune response to said RSV F protein; and

formulating said vector as a vaccine for in vivo administration.

19. A vaccine produced by the method of claim 18.

20. A plasmid vector, comprising:

a first nucleotide sequence encoding a truncated RSV F protein lacking a transmembrane region,

a promoter sequence operatively coupled to said nucleotide sequence for expression of said truncated RSV F protein, and

a second nucleotide sequence comprising a pair of splice sites to prevent aberrant mRNA splicing.

21. The vector of claim 20, wherein said promoter sequence is an immediate early cytomegalovirus promoter.

22. The vector of claim 20 wherein said first nucleotide sequence has SEQ ID NO:3.

23. The vector of claim 20 wherein said first nucleotide sequence encodes a truncated RSV F protein having an amino acid sequence of SEQ ID NO:4.

24. The vector of claim 20 wherein said further nucleotide sequence is located between said first nucleotide sequence and said promoter sequence.

25. The vector of claim 20 which is plasmid pXL2 as shown In FIG. 5.

26. The vector of claim 20 which is plasmid pXL4 as shown in FIG. 7.

27. The vector of claim 24 wherein said further nucleotide sequence is that of rabbit .beta.-globin intron II.

28. A method of immunizing a host against disease caused by infection with respiratory syncytial virus (RSV), which comprises administering to said host an effective amount of a plasmid vector comprising a nucleotide sequence having SEQ ID NO:3 or encoding a truncated RSV F protein lacking a transmembrane region and having SEQ ID NO:4, and a promoter sequence operatively coupled to said nucleotide sequence for expression of said truncated RSV F protein in said host.

29. The method of claim 28 wherein said host is a human.

30. The method of claim 29 wherein said promoter sequence is an immediately early cytomegalovirus promoter.

31. The method of claim 29 wherein said vector further comprises a further nucleotide sequence comprising a pair of splice sites to prevent mRNA splicing.

32. The method of claim 29 wherein said vector is selected from the group consisting of plasmid pXL2 as shown in FIG. 5 and plasmid pXL4 as shown in FIG. 7.

33. The method of claim 31 wherein said further nucleotide sequence is located between said nucleotide sequence and said promoter sequence.

34. The method of claim 33 wherein said further nucleotide sequence is that of rabbit .beta.-globin intron II.

35. A method of using a nucleotide sequence encoding a truncated RSV F protein lacking a transmembrane region to produce an immune response in a host, which comprises:

isolating said nucleotide sequence,

operatively linking said nucleotide expression to a promoter sequence to produce a plasmid vector wherein said promoter sequence directs expression of said truncated RSV F protein when said vector is introduced into a host to produce an immune response to said truncated RSV F protein; and

introducing said plasmid vector into the host.

36. The method of claim 35 wherein said host is a human host.

37. The method of claim 36 wherein said promoter sequence is an immediate early cytomegalovirus promoter.

38. The method of claim 36 wherein said nucleotide sequence has SEQ ID NO:3.

39. The method of claim 36 wherein said nucleotide sequence encodes a truncated RSV F protein having an amino acid sequence of SEQ ID NO:4.

40. The method of claim 36 wherein said vector further comprises a further nucleotide sequence comprising a pair of splice sites to prevent aberrant mRNA splicing.

41. The method of claim 36 wherein said plasmid vector is plasmid pXL2 shown in FIG. 5.

42. The method of claim 36 wherein said plasmid vector is plasmid pXL4 shown in FIG. 7.

43. The method of claim 40 wherein said further nucleotide sequence is located between said nucleotide sequence and said promoter sequence.

44. The method of claim 43 wherein said further nucleotide sequence is that of rabbit .beta.-globin intron II.

Details for Patent 6,022,864

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 06/04/1986 ⤷  Try a Trial 2015-06-07
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2015-06-07
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2015-06-07
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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