Modified Oncolytic Viruses

Abstract

The present invention provides an isolated selected Picornavirus capable of lytically infecting or inducing apoptosis in a cell substantially in the absence of intercellular adhesion molecule-1 (ICAM-1), and methods for treating subjects.

1 . A pharmaceutical composition comprising an isolated Picornavirus capable of lytically infecting or inducing apoptosis in a cell substantially in the absence of intercellular adhesion molecule-1 (ICAM-1), together with a suitable pharmaceutically acceptable excipient or diluent. 
     
     
         2 - 41 . (canceled) 
     
     
         42 . The pharmaceutical composition according to  claim 1 , wherein the selected Picornavirus is capable of lytically infecting a cell through decay-accelerating factor (DAF) on the cell. 
     
     
         43 . The pharmaceutical composition according to  claim 1 , wherein the Picornavirus is selected from the group consisting of prototype and clinically isolated strains of enteroviruses including Coxsackievirus, Echovirus, Poliovirus, unclassified enteroviruses, Rhinovirus, Paraechovirus, Hepatovirus, and Cardiovirus. 
     
     
         44 . The pharmaceutical composition according to  claim 1 , wherein the Picornavirus is a Coxsackievirus. 
     
     
         45 . The pharmaceutical composition according to  claim 44 , wherein the Coxsackievirus is type A. 
     
     
         46 . The pharmaceutical composition according to  claim 44 , wherein the Coxsackievirus is Coxsackievirus A21. 
     
     
         47 . The pharmaceutical composition according to  claim 1 , wherein the Picornavirus is an Echovirus. 
     
     
         48 . The Picornavirus according to  claim 47 , wherein the Echovirus is Echovirus 1, 6, 7, 8, 11, 12, 13 or 29. 
     
     
         49 . The Picornavirus according to  claim 1 , wherein the Picornavirus is a Poliovirus. 
     
     
         50 . The pharmaceutical composition according to  claim 1 , wherein the Picornavirus is bioselected by passaging a Picornavirus not capable of lytically infecting a cell without ICAM-1 in a DAF-expressing cell line without ICAM-1 and recovering the selected Picornavirus which is capable of lytically infecting a cell without ICAM-1. 
     
     
         51 . The pharmaceutical composition according to  claim 1 , wherein the Picornavirus is altered, mutated or modified, such as by site directed mutagenesis or passage in a cell where access to ICAM-1 is blocked by use of an anti-ICAM-1 antibody. 
     
     
         52 . The pharmaceutical composition according to  claim 1 , wherein the selected Picornavirus has an alteration in one or more capsid proteins compared with wild-type virus. 
     
     
         53 . The pharmaceutical composition according to  claim 52 , wherein the Picornavirus is a Coxsackievirus comprising an alteration in a capsid protein selected from VP1, VP2 and VP3. 
     
     
         54 . The pharmaceutical composition according to  claim 53 , wherein the mutation is selected from one or more of VP3 R96H; VP3 E101A; VP3 A239S; VP2S164L and VP2 V209. 
     
     
         55 . The pharmaceutical composition according to  claim 1 , wherein the selected Picornavirus comprises a capsid protein encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:7. 
     
     
         56 . The pharmaceutical composition according to  claim 1 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus which comprises:
 (i) a capsid protein encoded by the nucleotide sequence of SEQ ID NO: 7, or   (ii) a variant capsid protein encoded by a variant of the nucleotide sequence of SEQ ID NO: 7, the variant capsid protein comprising:
 (a) VP3 H96 or a conservative variation thereof wherein the conservative variation includes K, or 
 (b) VP3 A101 or a conservative variation thereof wherein A101 is substituted by an amino acid selected from the group consisting of G, T, S, and M, or 
 (c) both (a) and (b). 
   
     
     
         57 . The pharmaceutical composition according to  claim 1 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus in the form of CVA21-DAFv comprising a capsid protein encoded by the nucleotide sequence as set forth in SEQ ID NO: 7. 
     
     
         58 . The pharmaceutical composition according to  claim 1 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus which comprises:
 (i) a capsid protein having the amino acid sequence as set forth in SEQ ID NO: 8, or   (ii) a capsid protein that is a variant of the amino acid sequence set forth in SEQ ID NO: 8, the variant comprising:
 (a) a conservative substitution of VP3 H96 being VP3 H96K, or 
 (b) a conservative substitution of VP3 A101 selected from the group consisting of VP3 A101G, VP3 A101T, VP3 A101S, and VP3 A101M, or 
 (c) both (a) and (b). 
   
     
     
         59 . The pharmaceutical composition according to  claim 1 , wherein the cell is a neoplasm. 
     
     
         60 . The pharmaceutical composition according to  claim 59 , wherein the neoplasm is a DAF-expressing neoplasm. 
     
     
         61 . The pharmaceutical composition according to  claim 60 , wherein the neoplasm is selected from the group consisting of lung cancer, prostate cancer, colorectal cancer, thyroid cancer, renal cancer, adrenal cancer, liver cancer, leukemia, melanoma, pre-cancerous cells, oesophageal cancer, breast cancer, brain cancer, ovarian cancer, stomach and intestinal cancer. 
     
     
         62 . A method for treating a neoplasm in a mammal suffering from the neoplasm, the method comprising administering to the mammal an effective amount of an isolated Picornavirus capable of lytically infecting or inducing apoptosis in a cell substantially in the absence of intercellular adhesion molecule-1 (ICAM-1), under conditions which result in virus-mediated oncolysis of cells of the neoplasm. 
     
     
         63 . The method according to  claim 62 , wherein the neoplasm a DAF-expressing neoplasm. 
     
     
         64 . The method according to  claim 62 , wherein the neoplasm is selected from the group consisting of lung cancer, prostate cancer, colorectal cancer, thyroid cancer, renal cancer, adrenal cancer, liver cancer, leukemia, melanoma, pre-cancerous cells, oesophageal cancer, breast cancer, brain cancer, ovarian cancer, stomach and intestinal cancer. 
     
     
         65 . The method according to  claim 62 , wherein the Picornavirus is selected from the group consisting of prototype and clinically isolated strains of enteroviruses including Coxsackievirus, Echovirus, Poliovirus, unclassified enteroviruses, Rhinovirus, Paraechovirus, Hepatovirus, and Cardiovirus. 
     
     
         66 . The method according to  claim 62 , wherein the Coxsackievirus is type A. 
     
     
         67 . The method according to  claim 62 , wherein the Coxsackievirus is Coxsackievirus A21. 
     
     
         68 . The method according to  claim 62 , wherein the selected Picornavirus has an alteration in one or more capsid proteins compared with wild-type virus. 
     
     
         69 . The method according to  claim 68 , wherein the Picornavirus is a Coxsackievirus comprising an alteration in a capsid protein selected from VP1, VP2 and VP3. 
     
     
         70 . The method according to  claim 69 , wherein the mutation is selected from one or more of VP3 R96H; VP3 E101A; VP3 A239S; VP2S164L and VP2 V209. 
     
     
         71 . The method according to  claim 62 , wherein the selected Picornavirus comprises a capsid protein encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:7. 
     
     
         72 . The method according to  claim 62 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus which comprises:
 (i) a capsid protein encoded by the nucleotide sequence of SEQ ID NO: 7, or   (ii) a variant capsid protein encoded by a variant of the nucleotide sequence of SEQ ID NO: 7, the variant capsid protein comprising:
 (a) VP3 H96 or a conservative variation thereof wherein the conservative variation includes K, or 
 (b) VP3 A101 or a conservative variation thereof wherein A101 is substituted by an amino acid selected from the group consisting of G, T, S, and M, or 
 (c) both (a) and (b). 
   
     
     
         73 . The method according to  claim 62 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus in the form of CVA21-DAFv comprising a capsid protein encoded by the nucleotide sequence as set forth in SEQ ID NO: 7. 
     
     
         74 . The method according to  claim 62 , wherein the isolated Picornavirus is an isolated selected Coxsackie A21 virus which comprises:
 (i) a capsid protein having the amino acid sequence as set forth in SEQ ID NO: 8, or   (ii) a capsid protein that is a variant of the amino acid sequence set forth in SEQ ID NO: 8, the variant comprising:
 (a) a conservative substitution of VP3 H96 being VP3 H96K, or 
 (b) a conservative substitution of VP3 A101 selected from the group consisting of VP3 A101G, VP3 A101T, VP3 A101S, and VP3 A101M, or 
 (c) both (a) and (b).