Novel oligonucleotide conjugates and use thereof

A method for preparing a ligand-conjugated double-helix oligo RNA structure, the method comprising the steps of:

• (1) synthesizing a single-stranded RNA on a solid support having a functional group-hydrophilic material bonded thereto;
• (2) covalently bonding a hydrophobic material to the 5′ end of the single-stranded RNA having the functional group-hydrophilic material bonded thereto;
• (4) separating the functional group-RNA-polymer structure and a separately synthesized complementary single-stranded RNA from the solid support;
• (5) bonding a ligand to the end of the hydrophilic material by the functional group;
• and
• (6) annealing the ligand-bonded RNA-polymer structure with the complementary single-stranded RNA to form a double-stranded RNA structure.

A method for preparing a double-helix oligo RNA structure, the method comprising the steps of:

• (1) synthesizing a single-stranded RNA on a solid support;
• (2) covalently bonding a hydrophilic material to the 5′ end of the single-stranded RNA;
• (3) bonding a ligand to the hydrophilic material bonded to the single-stranded RNA;
• (4) separating the ligand-bonded, RNA-hydrophilic polymer structure and a separately synthesized complementary RNA-hydrophobic polymer structure from the solid support;
• and
• (5) annealing the ligand-bonded, RNA-hydrophilic polymer structure with the complementary RNA-hydrophobic polymer structure to form a double-stranded structure,
• wherein the preparation method comprises, between steps (1) to (4), a step of synthesizing a single-stranded RNA complementary to the single-stranded RNA of step (1), and then covalently bonding a hydrophobic material to the synthesized single-stranded RNA to synthesize a single-stranded RNA-hydrophobic polymer structure.

A method for preparing a ligand-bonded double-helix oligo RNA structure, the method comprising the steps of:

• (1) synthesizing a single-stranded RNA on a solid support having a functional group bonded thereto;
• (2) covalently bonding a hydrophilic material to the material obtained in step (1);
• (3) covalently bonding a ligand to the material obtained in step (2);
• (4) separating the material obtained in step (3) from the solid support;
• (5) covalently bonding a hydrophobic material to the material resulting from step (4) by the functional group bonded to the 3′ end;
• and
• (6) annealing the material resulting from step (5) with a complementary single-stranded RNA to form a double-strand RNA structure.

A method for preparing an ASO-polymer conjugate, the method comprising the steps of:

• (a) covalently bonding a hydrophilic material to a solid support;
• (b) synthesizing an ASO on the solid support comprising the hydrophilic material;
• (c) covalently bonding a hydrophobic material to the 5′ end of the ASO on the solid support;
• and
• (d) separating and purifying the resulting ASO-polymer conjugate from the solid support.

A method for preparing an ASO-polymer conjugate, the method comprising the steps of:

• (a) synthesizing an ASO on a solid support having a functional group bonded thereto;
• (b) covalently bonding a hydrophilic material to the 5′ end of the ASO;
• (c) separating the hydrophilic material-bonded ASO conjugate from the solid support;
• and
• (d) covalently bonding a hydrophobic material to the 3′ end of the ASO separated from the solid support.

A method for preparing an ASO-polymer conjugate comprising a ligand attached thereto, the method comprising the steps of:

• (a) bonding a hydrophilic material to a solid support having a functional group attached thereto;
• (b) synthesizing an ASO on the solid support having the functional group-hydrophilic material bonded thereto;
• (c) covalently bonding a hydrophobic material to the 5′ end of the ASO;
• (d) separating an ASO-polymer conjugate, obtained in step (c), from the solid support;
• and
• (e) bonding a ligand to the hydrophilic material of the ASO-polymer conjugate separated from the solid support.

A method for preparing an ASO-polymer conjugate comprising a ligand attached thereto, the method comprising the steps of:

• (a) synthesizing an ASO on a solid support having a functional group attached thereto;
• (b) covalently bonding a hydrophilic material to the end of the ASO;
• (c) covalently bonding a ligand to the ASO-hydrophilic material conjugate;
• (d) separating an ASO-hydrophilic polymer-ligand conjugate, which has the functional group attached thereto, from the solid support;
• and
• (e) covalently bonding a hydrophobic material to the 3′ end of the ASO of the conjugate separated from the solid conjugate.