Pharmaceutical composition for preventing or treating heart failure

Abstract

A pharmaceutical composition can be provide for treating or preventing heart failure. Additionally, siRNA and a vector expressing said siRNA can be provided that can be used in the pharmaceutical composition for treating or preventing heart failure. For example, a pharmaceutical composition can be provided for treating or preventing heart failure that contains a DNA sequence encoding RNA containing a sense strand sequence of consecutive 18 to 29 nucleotides from angiopoietin-like protein 2 (ANGPTL2) mRNA or the alternative splicing type RNA thereof and an antisense strand sequence complementary to the sense strand sequence under control of a promoter, and a pharmaceutically acceptable carrier.

1 - 27 . (canceled) 
     
     
         28 . A pharmaceutical composition for treating or preventing heart failure, comprising an expression vector containing a DNA sequence encoding RNA comprising a sense strand sequence of consecutive 18 to 29 nucleotides from angiopoietin-like protein 2 (ANGPTL2) mRNA or the alternative splicing type RNA thereof, and an antisense strand sequence complementary to the sense strand sequence under control of a promoter, and a pharmaceutically acceptable carrier,
 wherein the sense strand sequence and antisense strand sequence are adapted for use in preparing siRNA, which siRNA, when transduced into animal cell, suppresses expression of the angiopoietin-like protein 2 gene in the cell and has a silencing effect on the angiopoietin-like protein 2 gene.   
     
     
         29 . The pharmaceutical composition according to claim  1 , wherein the DNA sequence comprises the sense strand sequence, the antisense strand sequence and a sequence encoding a hairpin type RNA composed of a single strand loop sequence (hairpin sequence) bonding the sense strand sequence and the antisense strand sequence via a covalent bond,
 wherein the DNA sequence is adapted for use such that, when and the hairpin sequence is processed by Dicer, an intracellular RNase, siRNA comprising the sense strand sequence and the antisense strand sequence is formed.   
     
     
         30 . The pharmaceutical composition according to  claim 28 , wherein the expression vector has delivery selectivity into cardiomyocytes. 
     
     
         31 . The pharmaceutical composition according to  claim 29 , wherein the expression vector has delivery selectivity into cardiomyocytes. 
     
     
         32 . The pharmaceutical composition according to  claim 28 , wherein the composition is prepared to be administered into myocardium. 
     
     
         33 . The pharmaceutical composition according to  claim 29 , wherein the composition is prepared to be administered into myocardium. 
     
     
         34 . The pharmaceutical composition according to  claim 28 , wherein the sense strand sequence is any one of SEQ ID NO. 2 to SEQ ID NO. 8, or a sequence obtained by substitution, deletion or addition of one base in the nucleotide sequence: 
       
         
           
                 
                 
               
                     
                   SEQ ID NO. 2: 
                 
                     
                   GGAACAUUGACGGCGAAUA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 3: 
                 
                     
                   GAGAGUUCAUUUACCUAAA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 4: 
                 
                     
                   GGCUCUUACUCACUCAAGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 5: 
                 
                     
                   GGCAUUGUGAGCGAGGUGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 6: 
                 
                     
                   GCCAUUACCGGAGCCGCUA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 7: 
                 
                     
                   GUUUCCGCCUGGAACCUGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 8: 
                 
                     
                   GAAACUGUGCCCACUACCA. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         35 . The pharmaceutical composition according to  claim 28 , wherein the DNA sequence comprises a nucleotide sequence of any one of SEQ ID NO. 9 to SEQ ID NO. 15, or a sequence obtained by substitution, deletion or addition of one base in the nucleotide sequence: 
       
         
           
                 
                 
               
                     
                   SEQ ID NO. 9: 
                 
                     
                   GGAACATTGACGGCGAATA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 10: 
                 
                     
                   GAGAGTTCATTTACCTAAA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 11: 
                 
                     
                   GGCTCTTACTCACTCAAGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 12: 
                 
                     
                   GGCATTGTGAGCGAGGTGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 13: 
                 
                     
                   GCCATTACCGGAGCCGCTA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 14: 
                 
                     
                   GTTTCCGCCTGGAACCTGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 15: 
                 
                     
                   GAAACTGTGCCCACTACCA. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         36 . The pharmaceutical composition according to  claim 28 , wherein the expression vector is a plasmid or a viral vector. 
     
     
         37 . The pharmaceutical composition according to  claim 28 , wherein the viral vector is an adenoviral vector, an adeno-associated viral vector, a lentiviral vector or a retroviral vector. 
     
     
         38 . The pharmaceutical composition according to  claim 36 , wherein the viral vector is an adeno-associated virus serotype 6 (AAV6) vector. 
     
     
         39 . A pharmaceutical composition for treating or preventing heart failure, comprising siRNA comprising a sense strand sequence of consecutive 18 to 29 nucleotides from angiopoietin-like protein 2 (ANGPTL2) mRNA or alternative splicing type RNA thereof and an antisense strand sequence complementary to the sense strand sequence, and along with a pharmaceutically acceptable carrier,
 wherein siRNA containing the sense strand sequence and the antisense strand sequence is adapted for use such that when the siRNA is transduced into an animal cell, expression of the angiopoietin-like protein 2 gene is suppressed and a silencing effect on the angiopoietin-like protein 2 gene is produced.   
     
     
         40 . The pharmaceutical composition according to  claim 39 , wherein the siRNA targets the ANGPTL2 gene in cardiomyocytes of mammal. 
     
     
         41 . The pharmaceutical composition according to  claim 39 , wherein the sense strand sequence comprises a nucleotide sequence of any one of SEQ ID NO. 2 to SEQ ID NO. 8, or a sequence obtained by substitution, deletion or addition of one base in the nucleotide sequence: 
       
         
           
                 
                 
               
                     
                   SEQ ID NO. 2: 
                 
                     
                   GGAACAUUGACGGCGAAUA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 3: 
                 
                     
                   GAGAGUUCAUUUACCUAAA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 4: 
                 
                     
                   GGCUCUUACUCACUCAAGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 5: 
                 
                     
                   GGCAUUGUGAGCGAGGUGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 6: 
                 
                     
                   GCCAUUACCGGAGCCGCUA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 7: 
                 
                     
                   GUUUCCGCCUGGAACCUGA 
                 
                     
                     
                 
                     
                   SEQ ID NO. 8: 
                 
                     
                   GAAACUGUGCCCACUACCA. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         42 . A method for determining whether a subject is in need of treatment or prevention of heart failure with the pharmaceutical composition according to  claim 28  by measuring expression of angiopoietin-like protein 2 (ANGPTL2) in blood derived from a dilated cardiomyopathy (DCM) patient,
 wherein blood in the aortic root (Ao) and blood in the coronary sinus (CS) of a dilated cardiomyopathy patient are separately evaluated for expression levels of angiopoietin-like protein 2 (ANGPTL2), 
 wherein when the expression level of angiopoietin-like protein 2 (ANGPTL2) in the blood of the coronary sinus (CS) is higher than the expression level in the blood of the aortic root (Ao), it is determined that the subject from which the blood is derived is in need of treatment or prevention of heart failure. 
 
     
     
         43 . The method according to  claim 42 , wherein when the expression level of angiopoietin-like protein 2 (ANGPTL2) in the blood of the coronary sinus (CS) is higher than the expression level in the blood of the aortic root (Ao), it is determined that the subject from which the blood is derived is a subject to which the pharmaceutical composition according to  claim 28  is to be administered. 
     
     
         44 . The method of  claim 42 , further comprising administering the pharmaceutical composition according to  claim 28  to a patient whose expression level of angiopoietin-like protein 2 (ANGPTL2) in the blood of the coronary sinus (CS) is higher than the expression level in the blood of the aortic root (Ao).