Amphiregulin gene-specific double-stranded oligonucleotide and composition for preventing and treating fibrosis-related diseases and respiratory diseases, comprising same

Abstract

The present invention relates to a double-stranded oligonucleotide which can highly specifically and efficiently inhibit an amphiregulin expression and, preferably, a double-stranded oligonucleotide comprising a sequence in the form of RNA/RNA, DNA/DNA or DNA/RNA hybrid, a double-stranded oligonucleotide structure comprising the double-stranded oligonucleotide, nanoparticles comprising the double-stranded oligonucleotide structure, and a fibrosis or respiratory disease preventive or therapeutic use thereof.

1 . An amphiregulin-specific double-stranded oligonucleotide comprising: a sense strand comprising any one sequence selected from the group consisting of SEQ ID NOs: 10, 11 and 12; and an antisense strand comprising a sequence complementary thereto. 
     
     
         2 . The amphiregulin-specific double-stranded oligonucleotide of  claim 1 , wherein the sense strand or the antisense strand consists of 19 to 31 nucleotides. 
     
     
         3 . The amphiregulin-specific double-stranded oligonucleotide of  claim 1 , wherein the oligonucleotide is siRNA, shRNA or miRNA. 
     
     
         4 . The amphiregulin-specific double-stranded oligonucleotide of  claim 1 , wherein the sense or antisense strand is independently DNA or RNA. 
     
     
         5 . The amphiregulin-specific double-stranded oligonucleotide of  claim 1 , wherein the sense strand or the antisense strand of the double-stranded oligonucleotide comprises a chemical modification. 
     
     
         6 . The amphiregulin-specific double-stranded oligonucleotide of  claim 5 , wherein the chemical modification is any one or more selected from the group consisting of:
 modification in which a hydroxyl (OH) group at the 2′ carbon position of a sugar structure in nucleotides is substituted with any one selected from the group consisting of methyl (—CH 3 ), methoxy (—OCH 3 ), amine (—NH 2 ), fluorine (—F), —O-2-methoxyethyl, —O-propyl, —O-2-methylthioethyl, —O-3-aminopropyl, —O-3-dimethylaminopropyl, —O—N-methylacetamido and —O— dimethylamidooxyethyl;   modification in which oxygen in a sugar structure in nucleotides is substituted with sulfur;   modification of a bond between nucleotides to any one bond selected from the group consisting of a phosphorothioate bond, a boranophosphophate bond and a methyl phosphonate bond; and   modification to PNA (peptide nucleic acid), LNA (locked nucleic acid) or UNA (unlocked nucleic acid).   
     
     
         7 . The amphiregulin-specific double-stranded oligonucleotide of  claim 1 ,
 wherein one or more phosphate groups are bound to the 5′ end of the antisense strand of the double-stranded oligonucleotide.   
     
     
         8 . An amphiregulin-specific double-stranded oligonucleotide structure comprising a structure represented by the following Structural Formula 1:
   A-X—R—Y—B  [Structural Formula (1)]
   wherein A represents a hydrophilic compound, B represents a hydrophobic compound, X and Y each independently represent a simple covalent bond or a linker-mediated covalent bond, and R represents the double-stranded oligonucleotide of  claim 1 .   
     
     
         9 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the oligonucleotide structure comprises a structure represented by the following Structural Formula (2): 
       
         
           
           
               
               
           
         
         wherein S and AS respectively represent the sense strand and the antisense strand of the double-stranded oligonucleotide of  claim 8 . 
       
     
     
         10 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 9 , wherein oligonucleotide structure comprises a structure represented by the following Structural Formula (3) or (4): 
       
         
           
           
               
               
           
         
         wherein A, B, X, Y, S and AS are as defined in  claim 9 , and 5′ and 3′ represent the 5′ end and 3′ end, respectively, of the sense strand of the double-stranded oligonucleotide. 
       
     
     
         11 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the hydrophilic compound is selected from the group consisting of polyethylene glycol (PEG), polyvinylpyrrolidone, and polyoxazoline 
     
     
         12 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the hydrophilic compound has a structure represented by the following Structural Formula (5) or (6):
   (A′ m -J) n   [Structural Formula (5)]
     (J-A′ m ) n   [Structural Formula (6)]
   wherein A′ represents a hydrophilic monomer, J represents a linker that connects a number (m) of hydrophilic monomers together or connects a number (m) of hydrophilic monomers with the double-stranded oligonucleotide, m is an integer ranging from 1 to 15, n is an integer ranging from 1 to 10, the hydrophilic monomer (A′) is any one compound selected from among the following compound (1) to compound (3), and the linker (J) is selected from the group consisting of —PO 3   − —, —SO 3 — and —CO 2 —:   
       
         
           
           
               
               
           
         
         wherein G is selected from the group consisting of O, S and NH; 
       
       
         
           
           
               
               
           
         
       
     
     
         13 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 12 , wherein the oligonucleotide structure has a structure represented by the following Structural Formula (7) or (8):
   (A′ m -J) n -X—R—Y—B  [Structural Formula (7)]
     (J-A′ m ) n -X—R—Y—B  [Structural Formula (8)]
   
     
     
         14 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the hydrophilic compound has a molecular weight of 200 to 10,000. 
     
     
         15 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the hydrophobic compound has a molecular weight of 250 to 1,000. 
     
     
         16 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 15 , wherein the hydrophobic compound is any one selected from the group consisting of a steroid derivative, a glyceride derivative, glycerol ether, polypropylene glycol, a C 12 -C 50  unsaturated or saturated hydrocarbon, diacylphosphatidylcholine, a fatty acid, a phospholipid, lipopolyamine, a lipid, tocopherol, and tocotrienol. 
     
     
         17 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 16 , wherein the steroid derivative is any one selected from the group consisting of cholesterol, cholestanol, cholic acid, cholesteryl formate, cholestanyl formate, and cholestanyl amine. 
     
     
         18 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 16 , wherein the glyceride derivative is any one selected from the group consisting of mono-glyceride, di-glyceride, and tri-glyceride. 
     
     
         19 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 8 , wherein the covalent bond represented by X and Y is either a non-degradable bond or a degradable bond. 
     
     
         20 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 19 , wherein the non-degradable bond is an amide bond or a phosphate bond. 
     
     
         21 . The amphiregulin-specific double-stranded oligonucleotide structure of  claim 19 , wherein the degradable bond is any one selected from the group consisting of a disulfide bond, an acid-degradable bond, an ester bond, an anhydride bond, a biodegradable bond, and an enzyme-degradable bond. 
     
     
         22 . A nanoparticle comprising the double-stranded oligonucleotide of  claim 8 . 
     
     
         23 . The nanoparticle of  claim 22 , wherein the nanoparticle is composed of a mixture of double-stranded oligonucleotide structures comprising double-stranded oligonucleotide having different sequence. 
     
     
         24 . A method for preventing or treating fibrosis or respiratory disease, comprising administering to a subject in need thereof, the double-stranded oligonucleotide of  claim 1 , or an amphiregulin-specific double-stranded oligonucleotide structure comprising said double-stranded oligonucleotide, represented by the following Structural Formula 1:
   A-X—R—Y—B  [Structural Formula (1)]
   
       wherein A represents a hydrophilic compound, B represents a hydrophobic compound, X and Y each independently represent a simple covalent bond or a linker-mediated covalent bond, and R represents said double-stranded oligonucleotide. 
     
     
         25 . A method for preventing or treating fibrosis or respiratory disease, comprising administering the nanoparticle of  claim 22  to a subject in need thereof. 
     
     
         26 . The method of  claim 24 , wherein the respiratory disease is any one selected from the group consisting of chronic obstructive disease (COPD), asthma, acute and chronic bronchitis, allergic rhinitis, cough, sputum, bronchitis, bronchiolitis, sore throat, tonsillitis, and laryngitis. 
     
     
         27 . The method of  claim 24 , wherein the fibrosis is any one selected from the group consisting of idiopathic pulmonary fibrosis (IPF), liver fibrosis, cirrhosis, myelofibrosis, myocardial fibrosis, renal fibrosis, pulmonary fibrosis, cardiac fibrosis, and radiation-induced fibrosis. 
     
     
         28 . The method of  claim 25 , wherein the respiratory disease is any one selected from the group consisting of chronic obstructive disease (COPD), asthma, acute and chronic bronchitis, allergic rhinitis, cough, sputum, bronchitis, bronchiolitis, sore throat, tonsillitis, and laryngitis. 
     
     
         29 . The method pharmaceutical composition of  claim 25 , wherein the fibrosis is any one selected from the group consisting of idiopathic pulmonary fibrosis (IPF), liver fibrosis, cirrhosis, myelofibrosis, myocardial fibrosis, renal fibrosis, pulmonary fibrosis, cardiac fibrosis, and radiation-induced fibrosis. 
     
     
         30 . A lyophilized formulation comprising a pharmaceutical composition for preventing or treating fibrosis or respiratory disease, said pharmaceutical composition comprising the double-stranded oligonucleotide of  claim 1 , or an amphiregulin-specific double-stranded oligonucleotide structure comprising said double-stranded oligonucleotide, represented by the following Structural Formula 1:
   A-X—R—Y—B  [Structural Formula (1)]
   
       wherein A represents a hydrophilic compound, B represents a hydrophobic compound, X and Y each independently represent a simple covalent bond or a linker-mediated covalent bond, and R represents said double-stranded oligonucleotide. 
     
     
         31 . A lyophilized formulation comprising a pharmaceutical composition for preventing or treating fibrosis or respiratory disease, said pharmaceutical composition comprising the nanoparticle of  claim 22 .