Vegfr-2 targeting dna vaccine for combination therapy

Abstract

The present invention relates to an attenuated strain of Salmonella comprising at least one copy of a DNA molecule comprising an expression cassette encoding a VEGF receptor protein, for use in the treatment of cancer, wherein the treatment further comprises the administration of at least one further anti-cancer agent. The present invention further relates to a pharmaceutical composition comprising an attenuated strain of Salmonella comprising at least one copy of a DNA molecule comprising an expression cassette encoding a VEGF receptor protein, wherein the pharmaceutical composition further comprises at least one further attenuated strain of Salmonella comprising at least one copy of a further DNA molecule comprising a further expression cassette encoding a tumor antigen or a tumor stroma antigen.

1 . An attenuated strain of  Salmonella  comprising at least one copy of a DNA molecule comprising an expression cassette encoding a VEGF receptor protein, for use in the treatment of cancer, wherein the treatment further comprises the administration of
 i) at least one DNA vaccine encoding a tumor antigen or a tumor stroma antigen, particularly selected from at least one further attenuated strain of  Salmonella  comprising at least one copy of a further DNA molecule comprising a further expression cassette encoding a tumor antigen or a tumor stroma antigen, or   ii) at least one checkpoint inhibitor, particularly selected from an antibody against PD-1, PD-L1 and CTLA4, or   iii) at least one engineered T-cell, comprising at least one tumor antigen binding protein and/or at least one tumor stroma antigen binding protein on its cell surface; or   iv) at least one bispecific antibody exhibiting binding specificity for one T-cell surface protein and for a tumor antigen or for a tumor stroma antigen; or   v) or any combination thereof.   
     
     
         2 . The attenuated strain of  Salmonella  for use according to  claim 1 , wherein the attenuated strain of  Salmonella  and the at least one further attenuated strain of  Salmonella  are of the species  Salmonella enterica , particularly wherein the attenuated strain of  Salmonella  and the at least one further attenuated strain of  Salmonella  are  Salmonella typhi  Ty21a. 
     
     
         3 . The attenuated strain of  Salmonella  for use according to  claim 1  or  2 , wherein the expression cassette and the further expression cassette are a eukaryotic expression cassette, particularly wherein the expression cassette and the further expression cassette comprise a CMV promoter. 
     
     
         4 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the VEGF receptor protein is selected from the group consisting of human VEGFR-2 having the amino acid sequence as found in SEQ ID NO 1 and a protein that shares at least about 80% sequence identity therewith, particularly wherein human VEGFR-2 has the amino acid sequence as found in SEQ ID NO 1. 
     
     
         5 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the DNA molecule and the further DNA molecule comprise the kanamycin antibiotic resistance gene, the pMB1 ori, and a CMV promoter, particularly wherein the DNA molecule and the further DNA molecule comprise the DNA sequence as found in SEQ ID NO 2. 
     
     
         6 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein said tumor antigen encoded by said further attenuated strain of  Salmonella  is selected from the group consisting of human Wilms' Tumor Protein (WT1) having the amino acid sequence as found in SEQ ID NO 3 and a protein that shares at least about 80% sequence identity therewith, human Mesothelin (MSLN) having the amino acid sequence as found in SEQ ID NO 4 and a protein that shares at least about 80% sequence identity therewith, human CEA having the amino acid sequence as found in SEQ ID NO 5 and a protein that shares at least about 80% sequence identity therewith, CMV pp65 having the amino acid sequence as found in SEQ ID NO 6 and a protein that shares at least about 80% sequence identity therewith, CMV pp65 having the amino acid sequence as found in SEQ ID NO 7 and a protein that shares at least about 80% sequence identity therewith, and CMV pp65 having the amino acid sequence as found in SEQ ID NO 8 and a protein that shares at least about 80% sequence identity therewith, particularly wherein human Wilms' Tumor Protein (WT1) has the amino acid sequence as found in SEQ ID NO 3, human Mesothelin (MSLN) has the amino acid sequence as found in SEQ ID NO 4, human CEA has the amino acid sequence as found in SEQ ID NO 5, and CMV pp65 has the amino acid sequence as found in SEQ ID NO 6, SEQ ID NO 7 or SEQ ID NO 8, and wherein said tumor stroma antigen encoded by said further attenuated strain of  Salmonella  is selected from the group consisting of human fibroblast activation protein (FAP). 
     
     
         7 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the attenuated strain of  Salmonella  is administered simultaneously with or prior to said at least one DNA vaccine encoding a tumor antigen or a tumor stroma antigen, said at least one checkpoint inhibitor, said at least one engineered T-cell and/or said at least one bispecific antibody. 
     
     
         8 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the treatment is accompanied by chemotherapy, radiotherapy or biological cancer therapy, particularly wherein the attenuated strain of  Salmonella  is administered before or during the chemotherapy or the radiotherapy treatment cycle or the biological cancer therapy, or before and during the chemotherapy or the radiotherapy treatment cycle or the biological cancer therapy. 
     
     
         9 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the attenuated strain of  Salmonella  and the at least one further attenuated strain of  Salmonella  are administered orally. 
     
     
         10 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the cancer is selected from colorectal cancer, pancreatic cancer, lung cancer, ovarian cancer, mesothelioma, acute myeloid leukemia, chronic myeloid leukemia, glioblastoma, gastric cancer, hepatocellular cancer, renal cell cancer, prostate cancer, and cervical cancer. 
     
     
         11 . The attenuated strain of  Salmonella  for use according to of any one of the preceding claims, wherein the single dose of the attenuated strain of  Salmonella  and the at least one further attenuated strain of  Salmonella  comprises from about 10 5  to about 10 11 , particularly from about 10 6  to about 10 10 , more particularly from about 10 6  to about 10 9 , more particularly from about 10 6  to about 10 8 , most particularly from about 10 6  to about 10 7  colony forming units (CFU). 
     
     
         12 . The attenuated strain of  Salmonella  for use according to any one of the preceding claims, wherein the treatment is individualized cancer immunotherapy comprising the step of assessing the expression pattern of and/or the pre-immune response against said tumor antigen in a patient. 
     
     
         13 . A pharmaceutical composition comprising an attenuated strain of  Salmonella  comprising at least one copy of a DNA molecule comprising an expression cassette encoding a VEGF receptor protein, wherein the pharmaceutical composition further comprises at least one further attenuated strain of  Salmonella  comprising at least one copy of a further DNA molecule comprising a further expression cassette encoding a tumor antigen or a tumor stroma antigen. 
     
     
         14 . The pharmaceutical composition according to  claim 13 , wherein the attenuated strain of  Salmonella  and the at least one further attenuated strain of  Salmonella  is  Salmonella typhi  Ty21a, wherein the expression cassette and the further expression cassette is a eukaryotic expression cassette, particularly comprising a CMV promoter, and wherein the VEGF receptor protein is selected from the group consisting of human VEGFR-2 having the amino acid sequence as found in SEQ ID NO 1 and a protein that shares at least about 80% sequence identity therewith, particularly wherein human VEGFR-2 has the amino acid sequence as found in SEQ ID NO 1. 
     
     
         15 . The pharmaceutical composition according to  claim 13  or  14 , for use as a medicament, particularly for use in the treatment of cancer.