METHODS AND COMPOSITIONS FOR RNAi-BASED CANCER TREATMENT

Abstract

The present invention generally concerns methods and nucleotide-based compositions for treating metastatic cancer that is associated with a primary tumor or a few tumors at the primary organ.

We claim: 
     
         1 . A millimeter-scale drug delivery device (DDD), comprising:
 A. a biodegradable polymeric matrix; and   B. a nucleotide-based agent that targets a gene, incorporated within said biodegradable polymeric matrix,   for inhibiting the development of metastases of one or more primary tumors, by inserting said DDD into said primary tumor(s).   
     
     
         2 . The DDD of  claim 1 , wherein said primary tumor is selected from a pancreatic cancer tumor, a prostate cancer tumor, cervical tumor, a brain tumor, a breast tumor, a bladder tumor, colon cancer tumor, and a lung cancer tumor. 
     
     
         3 . The DDD of  claim 1 , wherein said gene is selected from the group consisting of Snai1, Snai2, PAK1, PAK2, ZEB1, ZEB2, Twist1, Twist2, Goosecoid, SIX1, FOXC2, Prx1, FN1, VIMENTIN (VIM), N-CADHERIN, SERPINA3, CD70, IL13RA2, CD74, and TCF3. 
     
     
         4 . The DDD of  claim 1 , wherein said gene is a mutated K-ras gene. 
     
     
         5 . The DDD of  claim 4 , wherein said nucleotide-based agent is an siRNA that comprises a duplex region, and the nucleotide sequence of the sense strand of said duplex region consists of a sequence selected from SEQ ID No: 1-7: 
       
         
           
                 
                 
               
                     
                   (SEQ ID No: 1) 
                 
                     
                   GUUGGAGCUGAUGGCG, 
                 
                     
                     
                 
                     
                   (SEQ ID No: 2) 
                 
                     
                   GUUGGAGCUGUUGGCG, 
                 
                     
                     
                 
                     
                   (SEQ ID No: 3) 
                 
                     
                   GUUGGAGCUGCUGGCG, 
                 
                     
                     
                 
                     
                   (SEQ ID No: 4) 
                 
                     
                   GUUGGAGCUAGUGGCG, 
                 
                     
                     
                 
                     
                   (SEQ ID No: 5) 
                 
                     
                   GUUGGAGCUUGUGGCG, 
                 
                     
                     
                 
                     
                   (SEQ ID No: 6) 
                 
                     
                   GUUGGAGCUGGUGACG, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID No: 7) 
                 
                     
                   GUUGGAGCUGGUUGCG, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         followed by a sequence selected from
 i. UAGGCAAGAGUGCC (SEQ ID No: 8); and 
 ii. a 5′-fragment of 3-13 nucleotides inclusive of SEQ ID No: 8. 
 
       
     
     
         6 . The DDD of  claim 5 , wherein said nucleotide sequence of the sense strand of said duplex region consists of a sequence selected from GUUGGAGCUGAUGGCGUAG (SEQ ID No: 9), GUUGGAGCUGUUGGCGUAG (SEQ ID No: 10), GUUGGAGCUGCUGGCGUAG (SEQ ID No: 11), GUUGGAGCUAGUGGCGUAG (SEQ ID No: 12), GUUGGAGCUUGUGGCGUAG (SEQ ID No: 13), and GUUGGAGCUGGUGACGUAG (SEQ ID No: 14), and GUUGGAGCUGGUUGCGUAG (SEQ ID No: 15). 
     
     
         7 . The DDD of  claim 4 , wherein said DDD further comprises another RNAi agent directed against a metastasis-facilitating gene. 
     
     
         8 . The DDD of  claim 7 , wherein said metastasis-facilitating gene is selected from the group consisting Snai1, Snai2, PAK1, PAK2, ZEB1, ZEB2, Twist1, Twist2, Goosecoid, SIX1, FOXC2, Prx1, and FN1, VIMENTIN (VIM), N-CADHERIN, SERPINA3, CD70, IL13RA2, CD74, and TCF3 
     
     
         9 . The DDD of  claim 1 , wherein said nucleotide-based agent is an siRNA. 
     
     
         10 . The DDD of  claim 1 , further comprising an additive for modulating drug-polymer hydrophobic-hydrophilic interactions. 
     
     
         11 . The DDD of  claim 1 , wherein said DDD is 0.68 mm+/−0.45 mm in diameter and is 4.5+/−2.5 mm in length. 
     
     
         12 . The DDD of  claim 1 , wherein said DDD contains between 200-470 micrograms of RNAi agent per DDD. 
     
     
         13 . The DDD of  claim 1 , wherein said nucleotide-based agent is chemically modified with a modification selected from the group consisting of 2′-O-methyl (2′-OMe), 2′-O-(2-methoxyethyl) (MOE) and 2′-fluorine. 
     
     
         14 . The DDD of  claim 1 , wherein said nucleotide-based agent is conjugated to a molecule selected from the group consisting of a cholesterol moiety, spermine, hydrophobized hyaluronic acid-spermine conjugates (HHSCs), or alpha-tocopherol-vitamin E, or a cell-penetrating peptide; or complexed with a cationic molecule. 
     
     
         15 . The DDD of  claim 1 , wherein the polymer in said biodegradable matrix comprises polylactic acid (PLA) and polyglycolic acid (PGA), wherein said PLA and PGA are present in a ratio of between 75:25 and 95:5, inclusive; and said polymer has a molecular weight of greater than 35 KDa. 
     
     
         16 . The DDD of  claim 1 , wherein said DDD is coated with a coating comprising a biodegradable polymer. 
     
     
         17 . The DDD of  claim 3 , wherein the sequence of the sense strand of the Snai2 RNAi consists of the sequence 5′-guuugcaagaucugcggca-3′ (SEQ ID NO: 69), 5′-cuggucaagaagcauuuca-3′ (SEQ ID NO: 70), 5′-guuugcaagaucugcggcadTdT-3′ (SEQ ID NO: 83) or 5′-cuggucaagaagcauuucadTdT-3′ (SEQ ID NO: 104). 
     
     
         18 . The DDD of  claim 3 , wherein the sequence of the sense strand of the Twist RNAi consists of the sequence 5′-gucugcagcucucgcccaa-3′ (SEQ ID NO: 71), 5′-ggugugcguccagccguug-3′ (SEQ ID NO: 72), 5′-gucugcagcucucgcccaadTdT-3′ (SEQ ID NO: 106), or 5′-ggugugcguccagccguugdTdT-3′ (SEQ ID NO: 108). 
     
     
         19 . The DDD of  claim 3 , wherein the sequence of the sense strand of the Zeb1 RNAi consists of the sequence 5′-gacucgagcauuuagacac-3′ (SEQ ID NO: 73), 5′-cagguguaagcgcagaaag-3′ (SEQ ID NO: 74), 5′-gacucgagcauuuagacacdTdT-3′ (SEQ ID NO: 110), or 5′-cagguguaagcgcagaaagdTdT-3′ (SEQ ID NO: 112). 
     
     
         20 . The DDD of  claim 3 , wherein the sequence of the sense strand of the Zeb2 RNAi consists of the sequence 5′-gucauuagaagaggcguaa-3′ (SEQ ID NO: 75), 5′-cauuagaagaggcguaaca-3′ (SEQ ID NO: 76), 5′-gucauuagaagaggcguaadTdT-3′ (SEQ ID NO: 114), or 5′-cauuagaagaggcguaacadTdT-3′ (SEQ ID NO: 116). 
     
     
         21 . The DDD of  claim 3 , wherein the sequence of the sense strand of the TCF3 RNAi consists of the sequence 5′-cuccuggacuucagcauga-3′ (SEQ ID NO: 99), 5′-gcacuggccucgaucuacu-3′ (SEQ ID NO: 101), 5′-cuccuggacuucagcaugadTdT-3′ (SEQ ID NO: 131), or 5′-gcacuggccucgaucuacudTdT-3′ (SEQ ID NO: 133).