Herpesvirus with modified glycoprotein B

Patent metadata

Field	Value
Publication number	US-11007236-B2
Application number	US-201716307683-A
Country	US
Kind code	B2
Filing date	Jun 8, 2017
Priority date	Jun 9, 2016
Publication date	May 18, 2021
Grant date	May 18, 2021

Abstract

The present invention is directed to a recombinant herpesvirus comprising a heterologous polypeptide ligand capable of binding to a target molecule and fused to or inserted into glycoprotein B at specific sites. The herpesvirus may comprise more than one ligand, and the additional ligand(s) may be comprised by a modified glycoprotein D and/or modified glycoprotein H. This allows the herpesvirus to target a cell for therapeutic purposes, and a cell for virus production. The present invention further comprises a pharmaceutical composition comprising the herpesvirus, the herpesvirus for use in the treatment of a tumor, infection, degenerative disorder or senescence-associated disease, a nucleic acid and a vector coding for the gB, a polypeptide comprising the gB, and a cell comprising the herpesvirus, nucleic acid, vector or polypeptide. Moreover, a method for infecting a cell with the herpesvirus or for producing the herpesvirus is disclosed.

First claim

The invention claimed is:

1. A recombinant herpesvirus comprising a heterologous polypeptide ligand capable of binding to a target molecule and inserted into glycoprotein B (gB) present in the envelope of the herpesvirus, wherein the ligand has a length of 5 to 30 amino acids and is inserted at any amino acid within the N-terminal region spanning from amino acids 31 to 108 of gB according to SEQ ID NO: 1, or within a corresponding region of a homologous gB.

2. The herpesvirus of claim 1 , wherein the herpesvirus has the capability of binding to a cell expressing or binding the target molecule, fusing with the cell membrane, entering the cell, or killing the cell.

3. The herpesvirus according to claim 1 , wherein the target molecule is present on a diseased cell or on a cell present in cell culture.

4. The herpesvirus according to claim 3 , wherein the target molecule present on a diseased cell is a tumor-associated receptor selected from the group consisting of: a member of the EGF receptor family, HER2, EGFR, EGFRIII, or EGFR3 (ERBB3), EGFRvIII, or MET, FAP, PSMA, CXCR4, CEA, CADC, Mucins, Folate-binding protein, GD2, VEGF receptors 1 and 2, CD20, CD30, CD33, CD52, CD55, the integrin family, IGF1R, the Ephrin receptor family, the protein-tyrosine kinase (TK) family, RANKL, TRAILR1, TRAILR2, IL13Ralpha, UPAR, Tenascin, a member of the immune checkpoint family regulators, PD-1, PD-L1, CTL-A4, TIM-3, LAG3, or IDO, tumor-associated glycoprotein 72, ganglioside GM2, A33, Lewis Y antigen, and MUC1; or
wherein the target molecule present on a cell present in cell culture is an artificial molecule, an antibody, an antibody derivative or an antibody mimetic, or a single-chain antibody (scFv).

5. The herpesvirus according to claim 1 , wherein the ligand is a natural polypeptide or an artificial polypeptide.

6. The herpesvirus according to claim 1 , wherein
a) the target molecule is HER2,
b) the target molecule comprises the amino acid sequence of SEQ ID NO: 41, or
c) the ligand comprises the amino acid sequence of SEQ ID NO: 37.

7. The herpesvirus according to claim 1 , further comprising one or more ligands, wherein the one or more ligands are fused to or inserted into gB; or wherein the gB comprises a ligand capable of binding to a target molecule present on a cell present in cell culture and a ligand capable of binding to a target molecule present on a diseased cell.

8. The herpesvirus according to claim 1 , wherein the herpesvirus comprises a modified gD and/or a modified gH.

9. The herpesvirus according to claim 8 , wherein the gD is modified to have a deletion of
a) amino acids 30 to 38 of gD or a subset thereof,
b) a deletion of amino acid 30 or amino acid 38, or
c) a deletion of amino acid 30 and amino acid 38,
with regard to mature gD according to SEQ ID NO: 62 or a corresponding region of a homologous gD.

10. The herpesvirus according to claim 8 , wherein a heterologous polypeptide ligand is inserted into gD instead of
a) amino acids 30 to 38 or a subset thereof,
b) amino acid 30 or amino acid 38, or
c) amino acid 38 and amino acid 30 is deleted,
with regard to mature gD according to SEQ ID NO: 62 or a corresponding region of a homologous gD.

11. The herpesvirus according to claim 1 , wherein the herpesvirus encodes one or more molecules that stimulate(s) the host immune response against a cell.

12. A pharmaceutical composition comprising the herpesvirus according to claim 1 and a pharmaceutically acceptable carrier.

13. A nucleic acid molecule comprising a nucleic acid coding for the gB of the herpesvirus according to claim 1 , having inserted the ligand, or
a vector comprising said nucleic acid molecule, or
a polypeptide comprising said gB, having inserted the ligand, or
a cell comprising said herpesvirus, said nucleic acid molecule, said vector, or said polypeptide.

14. The herpesvirus according to claim 3 , wherein the diseased cell is a tumor cell, an infected cell, a degenerative disorder-associated cell, or a senescent cell.

15. The herpesvirus according to claim 3 , wherein the cultured cell is a Vero cell, a 293 cell, a 293T cell, a HEp-2 cell, a HeLa cell, a BHK cell, or a RS cell.

16. The herpesvirus according to claim 14 , wherein the tumor cell is a breast cancer cell, ovary cancer cell, stomach cancer cell, lung cancer cell, head and neck cancer cell, osteosarcoma cell, glioblastoma multiforme cell, or salivary gland tumor cell.

17. The herpesvirus according to claim 4 , wherein the scFv is capable of binding to a part of the GCN4 yeast transcription factor.

18. The herpesvirus according to claim 1 , wherein the ligand is capable of binding to a part of the GCN4 yeast transcription factor.

19. The herpesvirus according to claim 8 , wherein the gB comprises a ligand capable of binding to a target molecule present on a cell present in cell culture and the modified gD and/or the modified gH comprises a ligand capable of binding to a target molecule present on a diseased cell.

20. The pharmaceutical composition of claim 12 , further comprising one or more molecules that stimulate the host immune response against a cell.

Assignees

• Univ Bologna Alma Mater Studiorum

Inventors

• Campadelli Maria Gabriella
• Petrovic Biljana

Classifications

• C12N15/86

  Viral vectors · CPC title

• C07K14/005

  from viruses · CPC title

• C12N2710/16622

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

• C07K2319/00

  Fusion polypeptide · CPC title

• C12N2710/16632

  Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent · CPC title