Complexes of cytomegalovirus proteins

Patent metadata

Field	Value
Publication number	US-2016159864-A1
Application number	US-201314410461-A
Country	US
Kind code	A1
Filing date	Jun 29, 2013
Priority date	Jul 6, 2012
Publication date	Jun 9, 2016
Grant date	—

Abstract

This disclosure provides an isolated human cytomegalovirus (HCMV) membrane protein complex, wherein said complex comprises gH, gL and at least one more HCMV glycoprotein. In some embodiments the complex consists of gH, gL and gO. In other embodiments the complex consists of gH, gL, pUL128, pUL130 and pUL131A. The disclosure also provides processes for expressing and purifying said complexes, and subsequent uses of said complexes in immunogenic compositions and vaccines.

First claim

1 - 27 . (canceled)

28 . A composition comprising purified human Cytomegalovirus (HCMV) pentameric complex comprising: HCMV gH or pentamer-forming fragment thereof, HCMV gL or pentamer-forming fragment thereof, HCMV pUL128 or pentamer-forming fragment thereof, HCMV pUL130 or pentamer-forming fragment thereof, and HCMV pUL131A or pentamer-forming fragment thereof, wherein the purity of said pentamer is at least 80% of the total proteinaceous mass in said composition.

29 . The composition of claim 28 , wherein said gH, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs. 1-4, 6, 29, and 30.

30 . The composition of claim 29 , wherein said gH, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs. 1-4, 6, 29, and 30.

31 . The composition of claim 28 , wherein said pentamer-forming fragment of gH comprises the ectodomain of the gH.

32 . The composition of claim 28 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs. 7-9 and 31.

33 . The composition of claim 28 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs. 7-9 and 31.

34 . The composition of claim 28 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs. 13-15 and 33.

35 . The composition of claim 28 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs. 13-15 and 33.

36 . The composition of claim 28 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs. 16, 17, and 34.

37 . The composition of claim 28 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs. 16, 17, and 34.

38 . The composition of claim 28 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs. 18-20, and 35.

39 . The composition of claim 28 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs. 18-20, and 35.

40 . A pharmaceutical comprising the composition of claim 28 , and an adjuvant.

41 . A composition comprising:
(A) a single recombinant nucleic acid construct, comprising one or more nucleotide sequences encoding: HCMV gH or pentamer-forming fragment thereof, HCMV gL or pentamer-forming fragment thereof, HCMV pUL128 or pentamer-forming fragment thereof, HCMV pUL130 or pentamer-forming fragment thereof, and HCMV pUL131A or pentamer-forming fragment thereof; or (B) two recombinant nucleic acid constructs, first construct comprising one or more nucleotide sequences encoding gH or pentamer-forming fragment thereof, and HCMV gL or pentamer-forming fragment thereof, the second construct comprising one or more nucleotide sequences encoding pUL128 or pentamer-forming fragment thereof, HCMV pUL130 or pentamer-forming fragment thereof, and HCMV pUL131A or pentamer-forming fragment thereof.

42 . The composition of claim 41 , wherein said pentamer-forming fragment of gH lacks a transmembrane domain.

43 . The composition of claim 41 , wherein said pentamer-forming fragment of gH comprises the ectodomain of the gH.

44 . A host cell comprising the composition of claim 41 .

45 . The host cell of claim 44 , wherein said host cell is a mammalian host cell.

46 . A cell culture for producing HCMV pentameric complex, comprising the host cell of claim 44 and a growth medium, wherein the amount of said pentameric complex is at a level of more than 0.4 mg per litre of growth medium.

47 . A method of treating HCMV infection, comprising administering to a subject in need there of an immunologically effective amount of the composition of claim 28 .

Assignees

• Novartis Ag

Inventors

• Carfi Andrea
• Wen Yingxia

Classifications

• A61K39/12

  Viral antigens · CPC title

• A61K2039/55566

  Emulsions, e.g. Freund's adjuvant, MF59 · CPC title

• C12N2710/16151

  Methods of production or purification of viral material · CPC title

• C12N2710/16122

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

• C07K14/005 Primary

  from viruses · CPC title