Method of treating cancer

Abstract

A method of treating cancer may include administering a polyplex of a double stranded RNA and a polymeric conjugate. The polymeric conjugate may consist of a linear polyethyleneimine covalently linked to one or more polyethylene glycol (PEG) moieties. Each PEG moiety may be conjugated via a linker to a targeting moiety capable of binding to a cancer antigen.

1 .- 20 . (canceled) 
     
     
         21 . A polyplex of a double stranded RNA (dsRNA) and a polymeric conjugate, wherein said polymeric conjugate consists of a linear polyethyleneimine (LPEI) covalently linked to one or more polyethylene glycol (PEG) moieties, each PEG moiety being conjugated via a linker to a targeting moiety capable of binding to a cancer antigen, provided that the targeting moiety is not mouse EGF or the peptide of the sequence YHWYGYTPQNVI (GE11) (SEQ ID NO: 1). 
     
     
         22 . The polyplex of  claim 21 , wherein the dsRNA is polyinosinic-polycytidylic acid double stranded RNA (poly I:C), the polymeric conjugate consists of LPEI covalently linked to one PEG moiety (LPEI-PEG 1:1) or to three PEG moieties (LPEI-PEG 1:3), and the cancer antigen is epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER2) or prostate surface membrane antigen (PSMA). 
     
     
         23 . The polyplex of  claim 21 , wherein said one or more PEG moieties each independently forms —NH—CO— bond with said LPEI and a bond selected from —NH—CO—, —CO—NH—, —S—C—, —S—S—, —O—CO— or —CO—O— with said linker. 
     
     
         24 . The polyplex of  claim 23 , wherein each one of said one or more PEG moieties forms —NH—CO— bonds with said LPEI and said linker. 
     
     
         25 . The polyplex of  claim 21 , wherein said linker forms an —S—S—, NH—CO—, —CO—NH—, —S—C—, —O—CO—, —CO—O— or urea (—NH—CO—NH) bond with said targeting moiety. 
     
     
         26 . The polyplex of  claim 25 , wherein said linker is selected from —CO—R 2 -R x -R 3  or a peptide moiety consisting of 3 to 7 amino acid residues,
 wherein 
 R 2  is selected from (C 1 -C 8 )alkylene, (C 2 -C 8 )alkenylene, (C 2 -C 8 ) alkynylene, (C 6 -C 10 )arylene-diyl, or heteroarylenediyl; 
 R x  is absent or —S—; 
 R 3  is absent or of the formula 
 
       
         
           
           
               
               
           
         
         R 4  is selected from (C 1 -C 8 )alkylene, (C 2 -C 8 )alkenylene, (C 2 -C 8 ) alkynylene, (C 1 -C 8 )alkylene-(C 3 -C 8 )cycloalkylene, (C 2 -C 8 )alkenylene-(C 3 -C 8 )cycloalkylene, (C 2 -C 8 ) alkynylene-(C 3 -C 8 )cycloalkylene, (C 6 -C 10 ) arylene-diyl, heteroarylenediyl, (C 1 -C 8 )alkylene-(C 6 -C 10 )arylene-diyl, or (C 1 -C 8 )alkylene-heteroarylenediyl; 
         wherein each one of said (C 1 -C 8 )alkylene, (C 2 -C 8 )alkenylene, or (C 2 -C 8 ) alkynylene is optionally substituted by one or more groups each independently selected from halogen, —COR 5 , —COOR 5 , —OCOOR 5 , —OCON(R 5 ) 2 , —CN, —NO 2 , —SR 5 , —OR 5 , —N(R 5 ) 2 , —CON(R 5 ) 2 , —SO 2 R 5 , —SO 3 H, —S(═O)R 5 , (C 6 -C 10 )aryl, (C 1 -C 4 )alkylene-(C 6 -C 10 )aryl, heteroaryl, or (C 1 -C 4 )alkylene-heteroaryl, and further optionally interrupted by one or more identical or different heteroatoms selected from S, O or N, and/or at least one group each independently selected from —NH—CO—, —CO—NH—, —N(R 5 )—, —N(C 6 -C 10 aryl)-, (C 6 -C 10 )arylene-diyl, or heteroarylenediyl; and 
         R 5  is H or (C 1 -C 8 )alkyl. 
       
     
     
         27 . The polyplex of  claim 26 , wherein R 2  is selected from (C 1 -C 8 )alkylene, preferably (C 1 -C 4 )alkylene, optionally substituted by one or more groups each independently selected from halogen, —COH, —COOH, —OCOOH, —OCONH 2 , —CN, —NO 2 , —SH, —OH, —NH 2 , —CONH 2 , —SO 2 H, —SO 3 H, —S(═O)H, (C 6 -C 10 )aryl, (C 1 -C 4 )alkylene-(C 6 -C 10 )aryl, heteroaryl, or (C 1 -C 4 )alkylene-heteroaryl, and further optionally interrupted by one or more identical or different heteroatoms selected from S, O or N, and/or at least one group each independently selected from —NH—CO—, —CO—NH—, —NH—, —N(C 1 -C 8 alkyl)-, —N(C 6 -C 10 aryl)-, (C 6 -C 10 )arylene-diyl, or heteroarylenediyl. 
     
     
         28 . The polyplex of  claim 27 , wherein R 2  is selected from (C 1 -C 8 )alkylene, preferably (C 1 -C 4 )alkylene. 
     
     
         29 . The polyplex of  claim 26 , wherein R x  is —S—. 
     
     
         30 . The polyplex of  claim 26 , wherein R 3  is absent or 
       
         
           
           
               
               
           
         
       
       wherein R 4  is (C 1 -C 8 )alkylene-(C 3 -C 8 )cycloalkylene, preferably (C 1 -C 4 )alkylene-(C 5 -C 6 )cycloalkylene. 
     
     
         31 . The polyplex of  claim 30 , wherein
 R 2  is —CH 2 —CH 2 —;   R x  is —S—; and   R 3  is absent or   
       
         
           
           
               
               
           
         
       
       wherein R 4  is 
       
         
           
           
               
               
           
         
       
     
     
         32 . The polyplex of  claim 26 , wherein said peptide moiety is —(NH—(CH 2 ) 7 —CO)-Phe-Gly-Trp-Trp-Gly-Cys- (SEQ ID NO: 2) or —(NH—(CH 2 ) 7 —CO)-Phe-Phe-(NH—CH 2 —CH(NH 2 )—CO)-Asp-Cys- (SEQ ID NO: 3), linked via its mercapto group to said targeting moiety. 
     
     
         33 . The polyplex of  claim 21 , wherein the polymeric conjugate is a diconjugate of the formula (i)-(viii), linked to said targeting moiety/moieties: 
       
         
           
           
               
               
           
         
       
       
         
           
           
               
               
           
         
         wherein R 7  is 
       
       
         
           
           
               
               
           
         
       
     
     
         34 . The polyplex of  claim 33 , wherein
 (a) said targeting moiety is HER2 affibody, and said polymeric conjugate is of the formula 13(i), and the HER2 affibody is linked via a mercapto group thereof;   (b) said targeting moiety is HER2 affibody, and said polymeric conjugate is of the formula 13(v), and the HER2 affibody is linked via a mercapto group thereof;   (c) said targeting moiety is EGFR affibody, and said polymeric conjugate is of the formula 13(i), and the EGFR affibody is linked via a mercapto group thereof;   (d) said targeting moiety is EGFR affibody, and said polymeric conjugate is of the formula 13(v), and the EGFR affibody is linked via a mercapto group thereof;   (e) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(ii), wherein the hEGF is linked via an amino group thereof;   (f) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(vi), wherein the hEGF is linked via an amino group thereof;   (g) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(iii);   (h) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(vii);   (i) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(iv); or   (j) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(viii).   
     
     
         35 . The polyplex of  claim 34 , wherein said EGFR affibody is of the amino acid sequence as set forth in SEQ ID NO: 4 and said HER2 affibody is of the amino acid sequence as set forth in SEQ ID NO: 5. 
     
     
         36 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the polyplex of  claim 21 . 
     
     
         37 . The method of  claim 43 , wherein said cancer characterized by EGFR-overexpressing cells is selected from non-small-cell-lung-carcinoma, breast cancer, glioblastoma, head and neck squamous cell carcinoma, colorectal cancer, adenocarcinoma, ovary cancer, bladder cancer or prostate cancer, and metastases thereof. 
     
     
         38 . The method of  claim 37 , wherein said polyplex is selected from the group consisting of a polyplex in which
 (c) said targeting moiety is EGFR affibody, and said polymeric conjugate is of the formula 13(i), and the EGFR affibody is linked via a mercapto group thereof;   (d) said targeting moiety is EGFR affibody, and said polymeric conjugate is of the formula 13(v), and the EGFR affibody is linked via a mercapto group thereof;   (e) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(ii), wherein the hEGF is linked via an amino group thereof; and   (f) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(vi), wherein the hEGF is linked via an amino group thereof.   
     
     
         39 . The method of  claim 43 , wherein said cancer characterized by HER2-overexpressing cells is selected from breast cancer, ovarian cancer, stomach cancer, and aggressive forms of uterine cancer, such as uterine serous endometrial carcinoma. 
     
     
         40 . The method of  claim 39 , wherein said cancer characterized by HER2-overexpressing cells is Herceptin/trastusumab resistant cancer. 
     
     
         41 . The method of  claim 39 , wherein said polyplex is selected from the group consisting of a polyplex in which
 (a) said targeting moiety is HER2 affibody, and said polymeric conjugate is of the formula 13(i), and the HER2 affibody is linked via a mercapto group thereof;   (b) said targeting moiety is HER2 affibody, and said polymeric conjugate is of the formula 13(v), and the HER2 affibody is linked via a mercapto group thereof;   (e) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(ii), wherein the hEGF is linked via an amino group thereof; and   (f) said targeting moiety is hEGF, and said polymeric conjugate is of the formula 13(vi), wherein the hEGF is linked via an amino group thereof.   
     
     
         42 . The method of  claim 43 , wherein said cancer is prostate cancer and said polyplex is selected from the group consisting of a polyplex in which
 (g) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(iii);   (h) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(vii);   (i) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(iv); or   (j) said targeting moiety is HOOC(CH 2 ) 2 —CH(COOH)—NH—CO—NH—CH(COOH)—(CH 2 ) 2 —CO— (DUPA residue), and said polymeric conjugate is of the formula 13(viii).   
     
     
         43 . A method for treating a cancer selected from the group consisting of a cancer characterized by EGFR-overexpressing cells, a cancer characterized by HER2-overexpressing cells and prostate cancer, said method comprising administering to a subject in need the polyplex according to claim  1 . 
     
     
         44 . The method of  claim 43  wherein the polyplex is administered in combination with immune cells. 
     
     
         45 . The method of  claim 44 , wherein said immune cells are tumor-infiltrating T-cells (T-TILs), tumor specific engineered T-cells, or peripheral blood mononuclear cells (PBMCs).