Recombinant rhabdovirus encoding for CCL21
The present invention relates to the field of oncolytic viruses and in particular to a recombinant rhabdovirus, such as vesicular stomatitis virus encoding in its genome for a CCL21 protein. The invention is further directed to the use of the recombinant virus in the treatment of cancer, and also to methods for producing such viruses.
1. A recombinant vesicular stomatitis virus encoding in its genome at least one human CCL21 protein having a c-terminal truncation wherein said truncated CCL21 protein comprises the amino acids 1-79, 1-81, 1-88, or 1-91, as shown in SEQ ID NO:2 and having at least 80% identity to SEQ ID NO:3, and wherein the gene coding for the glycoprotein G of the recombinant vesicular stomatitis virus is replaced by the gene coding for the glycoprotein GP of Lymphocyte choriomeningitis virus (LCMV), and/or the glycoprotein G is replaced by the glycoprotein GP of LCMV.
2. The recombinant vesicular stomatitis virus according to claim 1 , wherein said genome encodes for a plasmin processed truncated human CCL21 protein consisting of the amino acids 1-79, 1-81, 1-88, or 1-91, as shown in SEQ ID NO:2.
3. The recombinant vesicular stomatitis virus according to claim 1 , wherein said genome encodes for a c-terminally truncated human CCL21 protein consisting of SEQ ID NO:3.
4. The recombinant vesicular stomatitis virus according to claim 1 , wherein the said genome encodes for a truncated human CCL21 protein comprising SEQ ID NO:3 or having at least 96% identity to SEQ ID NO:3.
5. The recombinant vesicular stomatitis virus according to claim 1 , wherein said truncated human CCL21 protein has at least 80% identity to SEQ ID NO:4.
6. The recombinant vesicular stomatitis virus according to claim 1 , wherein the sequence encoding said truncated human CCL21 protein further comprises a signal peptide sequence operably linked to said human CCL2 protein encoding sequence.
7. The recombinant vesicular stomatitis virus according to claim 6 wherein the sequence encoding said human truncated CCL21 protein operably linked to signal sequence comprises SEQ ID NO:5 or a sequence having at least 80% identity to SEQ ID NO:5.
8. The recombinant vesicular stomatitis virus according to claim 1 , encoding in its genome a vesicular stomatitis virus nucleoprotein (N) wherein the nucleoprotein (N) comprises an amino acid sequence as set forth in SEQ ID NO: 7 or at least 98% identical to SEQ ID NO: 7.
9. The recombinant vesicular stomatitis virus according to claim 1 , encoding in its genome a vesicular stomatitis virus phosphoprotein (P) wherein the phosphoprotein (P) comprises an amino acid sequence as set forth in SEQ ID NO: 8 or at least 98% identical to SEQ ID NO: 8.
10. The recombinant vesicular stomatitis virus according to claim 1 , encoding in its genome a vesicular stomatitis virus large protein (L) wherein the large protein (L) comprises an amino acid sequence as set forth in SEQ ID NO:9 or at least 98% identical to SEQ ID NO:9.
11. The recombinant vesicular stomatitis virus according to claim 1 , encoding in its genome a vesicular stomatitis virus matrix protein (M) wherein the matrix protein (M) comprises an amino acid sequence as set forth in SEQ ID NO:10 or at least 98% identical to SEQ ID NO:10.
12. The recombinant vesicular stomatitis virus according to claim 1 , encoding in its genome:
the vesicular stomatitis virus nucleoprotein (N) comprising an amino acid sequence as set forth in SEQ ID NO:7 or at least 98% identical to SEQ ID NO:7
the vesicular stomatitis virus phosphoprotein (P) comprising an amino acid sequence as set forth in SEQ ID NO:8 or at least 98% identical to SEQ ID NO:8
the vesicular stomatitis virus large protein (L) comprising an amino acid sequence as set forth in SEQ ID NO:9 or at least 98% identical to SEQ ID NO:9, and
the vesicular stomatitis virus matrix protein (M) comprising an amino acid sequence as set forth in SEQ ID NO:10 or at least 98% identical to SEQ ID NO:10.
13. The recombinant vesicular stomatitis virus according to claim 1 , which is replication-competent.
14. A recombinant vesicular stomatitis virus encoding in its genome a vesicular stomatitis virus nucleoprotein (N), large protein (L), phosphoprotein (P), matrix protein (M), glycoprotein (G) and at least one human truncated CCL21 protein comprising the amino acids 1-79, 1-81, 1-88, or 1-91, as shown in SEQ ID NO:2 and having at least 80% identity to SEQ ID NO:3, and further comprising a signal sequence operably linked to said truncated CCL21 protein,
wherein the gene coding for the glycoprotein G of the vesicular stomatitis virus is replaced by the gene coding for the glycoprotein GP of lymphocyte choriomeningitis virus (LCMV), and/or the glycoprotein G is replaced by the glycoprotein GP of LCMV, and wherein
the nucleoprotein (N) comprises an amino acid as set forth in SEQ ID NO:7 or a at least 98% identical to SEQ ID NO:7
wherein the phosphoprotein (P) comprises an amino acid as set forth in SEQ ID NO:8 or at least 98% identical to SEQ ID NO:8
wherein the large protein (L) comprises an amino acid as set forth in SEQ ID NO:9 or at least 98% identical to SEQ ID NO:9
the matrix protein (M) comprises an amino acid as set forth in SEQ ID NO:10 or at least 98% identical to SEQ ID NO:10.
15. A pharmaceutical composition, characterized in that the composition comprises a recombinant vesicular stomatitis virus according to claim 1 .
16. A method for the treatment of solid cancers, comprising administration of a recombinant vesicular stomatitis virus according to claim 1 or a pharmaceutical composition comprising said vesicular stomatitis virus.
17. The method according to claim 16 , wherein the solid cancer is selected from the list comprising: reproductive tumor, an ovarian tumor, a testicular tumor, an endocrine tumor, a gastrointestinal tumor, a pancreatic tumor, a liver tumor, a kidney tumor, a colon tumor, a colorectal tumor, a bladder tumor, a prostate tumor, a skin tumor, melanoma, a respiratory tumor, a lung tumor, a breast tumor, a head & neck tumor, a head and neck squamous-cell carcinoma (HNSCC) and a bone tumor.
18. The method according to claim 16 , wherein administration of the recombinant vesicular stomatitis virus or the pharmaceutical composition is intratumorally or intravenously.
19. The method according to claim 16 , wherein administration of the recombinant vesicular stomatitis virus or the pharmaceutical composition is at least once intratumorally and subsequently intravenously.
20. The method according to claim 19 , wherein the subsequent intravenous administration is given 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days or 31 days after the initial intratumoral administration.
21. A composition comprising a recombinant vesicular stomatitis virus according to claim 1 and further a PD-1 pathway inhibitor or a SMAC mimetic.
22. The composition according to claim 21 , wherein the PD-1 pathway inhibitor is an antagonistic antibody which is directed against PD-1 or PD-L1.
23. The composition according to claim 21 , wherein the SMAC mimetic is selected from the group consisting of any one of compounds 1 to 26:
or a pharmaceutically acceptable salt of one of these compounds.
24. The composition according to claim 21 , wherein the PD-1 pathway inhibitor is an antagonist selected from the group consisting of pembrolizumab, nivolumab, pidilizumab, atezolizumab, avelumab, durvalumab, PDR-001, PD1-1, PD1-2, PD1-3, PD1-4 and PD1-5.
25. A kit of parts comprising:
a) a recombinant vesicular stomatitis virus as defined in claim 1 , or a pharmaceutical composition comprising said recombinant vesicular stomatitis virus, and
b) a PD-1 pathway inhibitor or SMAC mimetic.
26. The method according to claim 16 , wherein the recombinant vesicular stomatitis virus, or the pharmaceutical composition is administered in combination with a PD-1 pathway inhibitor or a SMAC mimetic.
27. The method according to claim 26 , wherein the recombinant vesicular stomatitis virus, or the pharmaceutical composition is administered concomittantly, sequentially or alternately with the PD-1 pathway inhibitor or the SMAC mimetic.
28. The method according to claim 26 , wherein the SMAC mimetic is selected from the group consisting of any one of compounds 1 to 26 according to claim 23 or a pharmaceutically acceptable salt of one of these compounds.
29. The method according to claim 26 , wherein the PD-1 pathway inhibitor is selected from the group consisting of pembrolizumab, nivolumab, pidilizumab, atezolizumab, avelumab, durvalumab, PDR-001, PD1-1, PD1-2, PD1-3, PD1-4 and PD1-5.
30. The method according to claim 26 , wherein the recombinant vesicular stomatitis virus or the pharmaceutical composition is administered via a different administration route then the PD-1 pathway inhibitor or the SMAC mimetic.
31. The method according to claim 26 , wherein the recombinant vesicular stomatitis virus or the pharmaceutical composition are administered at least once intratumorally and the PD-1 pathway inhibitor or the SMAC mimetic is administered intravenously.
32. A virus producing cell, characterized in that the cell produces a recombinant vesicular stomatitis virus according claim 1 .
33. The virus producing cell of claim 32 , characterized in that the cell is a Vero cell, a HEK cell, a HEK293 cell, a Chinese hamster ovary cell (CHO), or a baby hamster kidney (BHK) cell.
34. A recombinant rhabdovirus encoding in its RNA genome at least one human CCL21 protein, wherein the RNA genome of the recombinant rhabdovirus comprises or consists of a coding sequence identical or at least 75%, identical to SEQ ID NO: 24.