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) a first transgene insertion site located between the start site of adenoviral E1b-19K and the start site of adenoviral E1b-55K, the transgene insertion site comprising:
 a first DNA sequence encoding a first 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 first DNA sequence is present in the first transgene insertion site and is operably linked to a first endogenous promoter, and 
 a second DNA sequence encoding a second 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 second DNA sequence is present in the second transgene insertion site and is operably linked to a second endogenous promoter, 
 wherein the first DNA sequence and the second DNA sequence are operably linked or the first polypeptide and the second polypeptide are operably linked, and 
   (b) a modified adenoviral E1a regulatory sequence, wherein at least one Pea3 binding site of the adenoviral E1a regulatory sequence is modified or deleted.   
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the first DNA sequence and the second DNA sequence are operably linked by an IRES element. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the first polypeptide and the second polypeptide are operably linked by a self-cleaving 2A peptide. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein a sequence between two Pea3 sites of the adenoviral E1a regulatory sequence is deleted. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the first DNA sequence or the second DNA sequence encodes the chimeric human IL-12, wherein the chimeric human IL-12 comprises a p40 polypeptide, a p35 polypeptide, and a linker polypeptide. 
     
     
         6 . The pharmaceutical composition of  claim 1 , wherein the first DNA sequence or the second DNA sequence encodes human IL-2. 
     
     
         7 . The pharmaceutical composition of  claim 1 , wherein the first DNA sequence or the second DNA sequence encodes human IL-7. 
     
     
         8 . The pharmaceutical composition of  claim 1 , wherein the first polypeptide comprises human IL-7 and the second polypeptide comprises chimeric human IL-12. 
     
     
         9 . A pharmaceutical composition comprising an effective amount of a recombinant adenoviral vector comprising:
 (a) a first transgene insertion site located in the adenoviral E3 region, the transgene insertion site comprising a first DNA sequence encoding a first 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 first DNA sequence is present in the first transgene insertion site and is operably linked to a first endogenous promoter, and   (b) a modified adenoviral E1a regulatory sequence, wherein at least one Pea3 binding site of the adenoviral E1a regulatory sequence is modified or deleted.   
     
     
         10 . The pharmaceutical composition of  claim 9 , further comprising in the transgene insertion site a second DNA sequence encoding a second 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 second DNA sequence is present in the second transgene insertion site and is operably linked to a second endogenous promoter. 
     
     
         11 . The pharmaceutical composition of  claim 9 , wherein a sequence between two Pea3 sites of the adenoviral E1a regulatory sequence is deleted. 
     
     
         12 . The pharmaceutical composition of  claim 10 , wherein the first DNA sequence or the second DNA sequence encodes the chimeric human IL-12, wherein the chimeric human IL-12 comprises a p40 polypeptide, a p35 polypeptide, and a linker polypeptide. 
     
     
         13 . The pharmaceutical composition of  claim 10 , wherein the first DNA sequence or the second DNA sequence encodes human IL-2. 
     
     
         14 . The pharmaceutical composition of  claim 10 , wherein the first DNA sequence or the second DNA sequence encodes human IL-7. 
     
     
         15 . The pharmaceutical composition of  claim 10 , comprising a truncation in the E3 12.5K coding region, the E3 7.1K coding region, the E3 gp19K, the E3 10.5, or a combination thereof. 
     
     
         16 . A method for treating a tumor in a human subject in need thereof, comprising administering to the human with a tumor a therapeutic amount of the pharmaceutical composition of  claim 1  by systemic or intratumor administration. 
     
     
         17 . The method of  claim 16 , further comprising treating the human subject with an anti-PD-1 antibody or an anti-PD-L1 antibody. 
     
     
         18 . The method of  claim 16 , further comprising treating the human subject with:
 (a) an agonist of a co-stimulatory signal selected from glucocorticoid-induced tumor necrosis factor receptor (GITR), Inducible T-cell co-stimulator (ICOS or CD278), OX40 (CD134), CD27, CD28, 4-IBB (CD137), CD40, lymphotoxin alpha (LT alpha), LIGHT (lymphotoxin-like, exhibits inducible expression, and competes with herpes simplex virus glycoprotein D for HVEM, a receptor expressed by T lymphocytes), CD226, cytotoxic and regulatory T cell molecule (CRT AM), death receptor 3 (DR3), lymphotoxin-beta receptor (LTBR), transmembrane activator and CAML interactor (TACI), B cell-activating factor receptor (BAFFR), and B cell maturation protein (BCMA), or   (b) an antagonist of an inhibitory molecule selected from cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4 or CD52), programmed cell death protein 1 (PD1 or CD279), B and T-lymphocyte attenuator (BTLA), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG3), T-cell membrane protein 3 (TIM3), CD 160, adenosine A2a receptor (A2aR), T cell immunoreceptor with immunoglobulin and ITIM domains (TIGIT), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), and CD 160.   
     
     
         19 . A method for treating a tumor in a human subject in need thereof, comprising administering to the human with a tumor a therapeutic amount of the pharmaceutical composition of  claim 9  by systemic or intratumor administration. 
     
     
         20 . The method of  claim 19 , further comprising treating the human subject with:
 (a) an agonist of a co-stimulatory signal selected from glucocorticoid-induced tumor necrosis factor receptor (GITR), Inducible T-cell co-stimulator (ICOS or CD278), OX40 (CD134), CD27, CD28, 4-IBB (CD137), CD40, lymphotoxin alpha (LT alpha), LIGHT (lymphotoxin-like, exhibits inducible expression, and competes with herpes simplex virus glycoprotein D for HVEM, a receptor expressed by T lymphocytes), CD226, cytotoxic and regulatory T cell molecule (CRT AM), death receptor 3 (DR3), lymphotoxin-beta receptor (LTBR), transmembrane activator and CAML interactor (TACI), B cell-activating factor receptor (BAFFR), and B cell maturation protein (BCMA), or   (b) an antagonist of an inhibitory molecule selected from cytotoxic T lymphocyte-associated antigen 4 (CTLA-4 or CD52), programmed cell death protein 1 (PD1 or CD279), B and I-lymphocyte attenuator (BTLA), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG3), T-cell membrane protein 3 (TIM3), CD 160, adenosine A2a receptor (A2aR), T cell immunoreceptor with immunoglobulin and ITIM domains (TIGIT), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), and CD 160.