Immunomodulatory oncolytic adenoviral vectors, and methods of production and use thereof for treatment of cancer

Abstract

Disclosed herein are compositions and methods for treating cancer in a subject. This involves administering an oncolytic virus containing a heterologous DNA sequence encoding one or more immunomodulatory and/or immunostimulatory polypeptide(s) of interest to the subject under conditions effective to enhance an anti-tumor immune response in the subject, and to treat cancer. It also relates to a method of enhancing the delivery to and distribution within a tumor mass of therapeutic viruses.

What is claimed is: 
     
         1 . A pharmaceutical composition comprising an effective amount of a recombinant adenoviral vector comprising: a transgene insertion site located between the start site of adenoviral E1b-19K and the start site of adenoviral E1b-55K, wherein a first DNA sequence and a second DNA sequence are each inserted into the transgene insertion site;
 wherein the first DNA sequence encodes a polypeptide selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, and a human OX40 ligand, and wherein the second DNA sequence encodes a polypeptide selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, and a human OX40 ligand;   wherein the adenoviral vector comprises a modified adenoviral E1a regulatory sequence wherein at least one Pea3 binding site, or a functional portion thereof, of the recombinant adenoviral vector is modified or deleted.   
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the adenoviral vector comprises an IRES element or encodes a self-cleaving 2A peptide sequence between the first DNA sequence and the second DNA sequence. 
     
     
         3 . The pharmaceutical composition of  claim 1 , comprising a modified E3 region. 
     
     
         4 . The pharmaceutical composition of  claim 1 , comprising an intact E3 region. 
     
     
         5 . The pharmaceutical composition of  claim 3 , further comprising a third DNA sequence inserted into the E3 region, wherein the third DNA sequence encodes a polypeptide selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, or a human OX40 ligand. 
     
     
         6 . The pharmaceutical composition of  claim 1 , wherein the adenoviral vector comprises a nucleic acid sequence at least 95% identical in an E3 region to vector d1327. 
     
     
         7 . The pharmaceutical composition of  claim 1 , wherein the chimeric human IL-12 polypeptide comprises a p40 polypeptide, a p35 polypeptide, and a linker polypeptide. 
     
     
         8 . The pharmaceutical composition of  claim 1 , wherein the chimeric human IL-12 polypeptide comprises a sequence as set forth in SEQ ID NO:46. 
     
     
         9 . The pharmaceutical composition of  claim 1 , formulated for systemic administration. 
     
     
         10 . The pharmaceutical composition of  claim 1 , formulated for intratumoral administration. 
     
     
         11 . A pharmaceutical composition comprising an effective amount of a recombinant adenoviral vector comprising:
 a. a first transgene insertion site located between the start site of adenoviral E1b-19K and the start site of adenoviral E1b-55K;   b. a second transgene insertion site located in adenoviral E3 region;   c. a first DNA sequence, present in the first transgene insertion site, encoding one or a plurality of polypeptides selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, and a human OX40 ligand; and   d. a second DNA sequence, present in the second transgene insertion site, encoding one or a plurality of polypeptides selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, and a human OX40 ligand,   wherein the adenoviral vector comprises a modified adenoviral E1a regulatory sequence.   
     
     
         12 . The pharmaceutical composition of  claim 11 , further comprising a third DNA sequence inserted into a third transgene insertion site, encoding one or a plurality of polypeptides selected from the group consisting of: a chimeric human IL-12, a human IL-7, an anti-CTLA-4 antibody, an IL-10Rtrap, a human CD70, a human IL-2 polypeptide, a human CD40 ligand, and a human OX40 ligand. 
     
     
         13 . The pharmaceutical composition of  claim 8 , wherein at least one of the first DNA sequence, the second DNA sequence, and the third DNA sequence independently comprises an IRES element and/or a self-cleaving 2A peptide. 
     
     
         14 . The pharmaceutical composition of  claim 11 , wherein at least one E1a regulatory sequence Pea3 binding site, or a functional portion thereof of the adenoviral vector, is modified or deleted. 
     
     
         15 . The pharmaceutical composition of  claim 11 , wherein a sequence between two Pea3 sites of the adenoviral vector is deleted. 
     
     
         16 . The pharmaceutical composition of  claim 11 , comprising a modified E3 region. 
     
     
         17 . The pharmaceutical composition of  claim 11 , wherein the adenoviral vector comprises a nucleic acid sequence at least 95% identical in an E3 region to vector d1327. 
     
     
         18 . The pharmaceutical composition of  claim 11 , wherein the chimeric human IL-12 polypeptide comprises a p40 polypeptide, a p35 polypeptide, and a linker polypeptide. 
     
     
         19 . A method for treating a tumor in a human subject in need thereof, comprising administering to the human with the tumor a therapeutic amount of the pharmaceutical composition of  claim 1  by systemic or intratumoral administration. 
     
     
         20 . A method for treating a tumor in a human subject in need thereof, comprising administering to the human with the tumor a therapeutic amount of the pharmaceutical composition of  claim 11  by systemic or intratumoral administration.