Recombinant oncolytic virus, synthetic dna sequence, and application thereof

Abstract

A recombinant oncolytic virus, a synthetic DNA sequence and applications of the virus are described. The recombinant oncolytic virus includes a genome and an exogenous DNA sequence inserted in the genome. The exogenous DNA sequence adapts to express a basic peptide fragment, to increase the environmental pH in a host infected by the recombinant oncolytic virus. More than 60% of amino acids in the basic peptide fragment are basic amino acids. The recombinant oncolytic virus and the synthetic DNA sequence of the disclosure are used to prepare an anti-tumor drug.

What is claimed is: 
     
         1 . A recombinant oncolytic virus, comprising: an oncolytic virus genome and an exogenous DNA sequence inserted in the oncolytic virus genome, the exogenous DNA sequence being adapted to express a basic peptide fragment and to increase an environmental pH in a host infected by the recombinant oncolytic virus. 
     
     
         2 . The virus of  claim 1 , wherein the oncolytic virus genome is a genome of herpes virus, Coxsackie virus, adenovirus, cowpox virus, measles virus, poliomyelitis virus, retrovirus, reovirus, respiratory syncytial virus, parvovirus H1, vesicular stomatitis virus, or Newcastle disease virus. 
     
     
         3 . The virus of  claim 1 , wherein the environmental pH in the host infected by the recombinant oncolytic virus is increased by 0.4 to 0.6. 
     
     
         4 . The virus of  claim 1 , wherein the basic peptide fragment comprises 4 to 10 amino acids. 
     
     
         5 . The virus of  claim 4 , wherein more than 60% of amino acids in the basic peptide fragment are basic amino acids. 
     
     
         6 . The virus of  claim 5 , wherein more than 80% of amino acids in the basic peptide fragment are basic amino acids. 
     
     
         7 . The virus of  claim 5 , wherein the basic amino acids are selected from arginine, lysine, and histidine. 
     
     
         8 . The virus of  claim 1 , wherein the basic peptide fragment is selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 3) 
                 
                     
                   Arg-Lys-Arg-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 5) 
                 
                     
                   Lys-Arg-Lys-Arg; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 7) 
                 
                     
                   Arg-Arg-Lys-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 9) 
                 
                     
                   Lys-Lys-Arg-Arg; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   Lys-Arg-Arg-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   Arg-Lys-Lys-Arg; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                   Arg-Arg-His-Lys-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                   Lys-His-Arg-Lys-His-Arg; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 19) 
                 
                     
                   Lys-His-Arg-Cys-Lys-Pro; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 21) 
                 
                     
                   Arg-Arg-His-Lys-Met-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 23) 
                 
                     
                   His-Arg-Lys-Cys-Arg-Lys; 
                 
                     
                   (SEQ ID NO: 25) 
                 
                     
                     
                 
                     
                   Lys-Arg-Trp-Arg-Lys-His-Arg; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 27) 
                 
                     
                   His-Lys-Gly-Arg-Lys-Cys-Arg-Val; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 29) 
                 
                     
                   Lys-Arg-Trp-His-Lys-Met-Arg-Lys-His; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                   His-Phe-Trp-Arg-Gln-Cys-Ala-Met-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 33) 
                 
                     
                   Tyr-Phe-Pro-Arg-His-Gln-Lys-Trp-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 35) 
                 
                     
                   Trp-Lys-Tyr-Arg-Gln-Ile-Ser-Thr-Cys; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 37) 
                 
                     
                   Arg-Lys-His-Lys-Met-Arg-Lys-Cys-His-Lys. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         9 . The virus of  claim 1 , wherein the recombinant oncolytic virus is Coxsackie virus B3 strain. 
     
     
         10 . The virus of  claim 9 , wherein the basic peptide fragment is selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 11) 
                 
                     
                   Lys-Arg-Arg-Lys; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 29) 
                 
                     
                   Lys-Arg-Trp-His-Lys-Met-Arg-Lys-His; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                   His-Phe-Trp-Arg-Gln-Cys-Ala-Met-Lys. 
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         11 . The virus of  claim 10 , wherein the recombinant oncolytic virus is a variant attenuated Coxsackie virus B3 strain comprising base mutations of T97C, G1180A, T1654C, T1756C, G2276A, A2685C, G2690A, C3120A, A3231G, G4327A, T5088C, A5270G, C7026T, and/or G7192A. 
     
     
         12 . The virus of  claim 10 , wherein the exogenous DNA sequence is inserted onto a pVAX1 vector. 
     
     
         13 . The virus of  claim 10 , wherein the basic peptide fragment is 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 31) 
                 
                     
                   His-Phe-Trp-Arg-Gln-Cys-Ala-Met-Lys. 
                 
             
                
                
               
            
           
         
       
     
     
         14 . The virus of  claim 10 , wherein the basic peptide fragment is 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 35) 
                 
                     
                   Trp-Lys-Tyr-Arg-Gln-Ile-Ser-Thr-Cys. 
                 
             
                
                
               
            
           
         
       
     
     
         15 . An anti-tumor drug, comprising the recombinant oncolytic virus of  claim 1 . 
     
     
         16 . The drug of  claim 15 , wherein the drug further comprises a checkpoint inhibitor. 
     
     
         17 . A method for treating a malignant tumor, comprising administering the anti-tumor drug of  claim 15  intravenously or locally to a patient in need thereof. 
     
     
         18 . The method of  claim 17 , wherein the malignant tumor is a solid tumor. 
     
     
         19 . The method of  claim 18 , wherein the malignant tumor is respiratory tract tumor, gastrointestinal tumor, endocrine tumor, or gynecological tumor.