Cytomegalovirus antigens and uses thereof

Patent metadata

Field	Value
Publication number	US-10167321-B2
Application number	US-201615544730-A
Country	US
Kind code	B2
Filing date	Jan 22, 2016
Priority date	Jan 22, 2015
Publication date	Jan 1, 2019
Grant date	Jan 1, 2019

Abstract

This disclosure provides modified cytomegalovirus (CMV) gL proteins and complexes comprising gL proteins. The modified gL proteins remain intact and are able to form complexes with other CMV proteins.

First claim

The invention claimed is:

1. A recombinant Human Cytomegalovirus (HCMV) gL protein, or a complex-forming fragment thereof, wherein said gL protein or fragment comprises a mutation at Protease Recognition Site residues 91-102 numbered according to SEQ ID NO: 1, wherein said mutation reduces protease cleavage at said Protease Recognition Site as compared to a control.

2. The gL protein or fragment of claim 1 , wherein said mutation comprises addition of one to five amino acid residues.

3. The gL protein or fragment of claim 2 , comprising an addition of two to five amino acid residues and wherein said two to five amino acid residues comprise both polar residue(s) and non-polar residue(s).

4. The gL protein or fragment of claim 2 , wherein said mutation comprises addition of one to five residues between residues corresponding to N97 and S98 of SEQ ID NO: 1.

5. The gL protein or fragment of claim 4 , wherein said mutation comprises addition of F, Q, FQ or QF between residues corresponding to N97 and S98 of SEQ ID NO: 1.

6. The gL protein or fragment of claim 1 , wherein said mutation comprises deletion of one to three amino acid residues.

7. The gL protein or fragment of claim 1 , wherein said mutation comprises deletion of a residue corresponding to the residue of SEQ ID NO: 1 selected from the group consisting of: V91, T92, P93, E94, A95, A96, N97, S98, V99, L100, L101, D102 and a combination thereof.

8. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting a residue with a corresponding residue from the gL protein of another herpes virus.

9. The gL protein or fragment of claim 8 , wherein said gL protein from another herpes virus is gL protein from HSV1, HSV2, VZV, EBV, PrV, or bovine herpesvirus 5.

10. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting a residue corresponding to the residue of SEQ ID NO: 1 selected from the group consisting of: A96, N97, S98, and a combination thereof.

11. The gL protein or fragment of claim 10 , wherein said mutation comprises substituting the residue corresponding to the A96 residue of SEQ ID NO: 1 with another non-polar residue and substituting the residues corresponding to the N97 and S98 residues of SEQ ID NO: 1 with another residue that comprises a small side chain.

12. The gL protein or fragment of claim 11 , wherein said residue corresponding to the A96 residue of SEQ ID NO: 1 is substituted to L or I, said residue corresponding to the N97 residue of SEQ ID NO: 1 is substituted to S or D, and said residue corresponding to the S98 residue of SEQ ID NO: 1 is substituted to G.

13. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to A96 of SEQ ID NO: 1 with a non-polar residue or with a residue that comprises a large side chain.

14. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to A96 of SEQ ID NO: 1 with I, L, V or S.

15. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to A95 of SEQ ID NO: 1 with R, L, E or N.

16. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to E94 of SEQ ID NO: 1 with A or L.

17. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to N97 of SEQ ID NO: 1 with a polar residue or a non-polar residue.

18. The gL protein or fragment of claim 17 , wherein said polar residue comprises a small side chain.

19. The gL protein or fragment of claim 17 , wherein said polar residue comprises a large side chain.

20. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to N97 of SEQ ID NO: 1 with S, D, E, A, T or Y.

21. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to S98 of SEQ ID NO: 1 with an amino acid residue with a small side chain.

22. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to S98 of SEQ ID NO: 1 with G, T, V, or I.

23. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to V99 of SEQ ID NO: 1 with I.

24. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to L100 of SEQ ID NO: 1 with F or V.

25. The gL protein or fragment of claim 1 , wherein said mutation comprises substituting the residue corresponding to L101 of SEQ ID NO: 1 with V or I.

26. The gL protein or fragment of claim 1 , wherein said gL protein or fragment comprises an Insert Region at the N-terminus of the Protease Recognition Site.

27. The gL protein or fragment of claim 1 , wherein said mutation comprises introducing a non-naturally occurring amino acid residue.

28. The gL protein or fragment of claim 1 , wherein said mutation comprises introducing an amino acid residue comprising a bulky side chain.

29. An HCMV complex comprising the recombinant gL protein or fragment of claim 1 .

30. The complex of claim 29 , wherein said complex is a pentameric complex comprising: gH or a pentamer-forming fragment thereof, gL or a pentamer-forming fragment thereof, pUL128 or a pentamer-forming fragment thereof, pUL130 or a pentamer-forming fragment thereof, and pUL131 or a pentamer-forming fragment thereof.

31. The complex of claim 30 , wherein said complex comprises a gH pentamer-forming fragment that lacks a transmembrane domain.

32. An HCMV complex comprising the recombinant gL protein or fragment of claim 10 .

33. The complex of claim 32 , wherein said complex is a pentameric complex comprising: gH or a pentamer-forming fragment thereof, gL or a pentamer-forming fragment thereof, pUL128 or a pentamer-forming fragment thereof, pUL130 or a pentamer-forming fragment thereof, and pUL131 or a pentamer-forming fragment thereof.

34. The complex of claim 33 , wherein said complex comprises a gH pentamer-forming fragment that lacks a transmembrane domain.

35. An isolated nucleic acid comprising a polynucleotide sequence encoding the recombinant gL protein or fragment of claim 1 .

36. An isolated nucleic acid comprising a polynucleotide sequence encoding the recombinant gL protein or fragment of claim 10 .

37. An isolated nucleic acid comprising a polynucleotide sequence encoding the complex of claim 29 .

38. An isolated nucleic acid comprising a polynucleotide sequence encoding the complex of claim 32 .

39. A vector comprising the isolated nucleic acid of claim 35 .

40. A vector comprising the isolated nucleic acid of claim 36 .

41. A vector comprising the isolated nucleic acid of claim 37 .

42. A vector comprising the isolated nucleic acid of claim 38 .

43. An isolated host cell comprising the isolated nucleic acid of claim 35 .

44. An isolated host cell comprising the isolated nucleic acid of claim 36 .

45. An isolated host cell comprising the isolated

Assignees

• Glaxosmithkline Biologicals Sa

Inventors

• Carfi Andrea
• Ciferri Claudio
• Xing Yi

Classifications

• A61P31/20

  for DNA viruses · CPC title

• A61K39/245 Primary

  Herpetoviridae, e.g. herpes simplex virus · CPC title

• C07K16/082

  Hepadnaviridae (F), e.g. hepatitis B virus · CPC title

• C12N2710/16122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K38/00

  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 title