LIVER CANCER RELATED GENES-SPECIFIC siRNA, DOUBLE-STRANDED OLIGO RNA MOLECULES COMPRISING THE siRNA, AND COMPOSITION FOR PREVENTING OR TREATING CANCER COMPRISING THE SAME

Patent metadata

Field	Value
Publication number	US-2016168573-A1
Application number	US-201414902808-A
Country	US
Kind code	A1
Filing date	Jul 9, 2014
Priority date	Jul 9, 2013
Publication date	Jun 16, 2016
Grant date	—

Abstract

There is provided a liver cancer related specific siRNA and high efficiency double-stranded oligo RNA molecules containing the same. The double-stranded oligo RNA molecules have a structure in which hydrophilic and hydrophobic compounds are conjugated to both ends of the double-stranded oligo RNA molecules by a simple covalent bond or a linker-mediated covalent bond in order to be efficiently delivered into cells and may be converted into nanoparticles in an aqueous solution by hydrophobic interactions of the double-stranded oligo RNA molecules. The siRNA contained in the double-stranded oligo RNA molecules may be liver cancer related genes, particularly Gankyrin or BMI-1 specific siRNA.
In addition, the present invention relates to a method of preparing the double-stranded oligo RNA molecules, and a pharmaceutical composition for preventing or treating cancer, particularly, liver cancer, containing the double-stranded oligo RNA molecules.

First claim

1 . A Gankyrin or BMI-1 specific siRNA comprising a sense strand comprising any one sequence selected from SEQ ID NOs. 1 to 200 and an antisense strand comprising a sequence complementary thereto.

2 . The siRNA of claim 1 , wherein the sense or antisense strand of the siRNA is composed of 19 to 31 nucleotides.

3 . The siRNA of claim 1 , composed of a sense strand comprising any one sequence selected from a group consisting of SEQ ID NOs. 1, 10, 13, 56, 99, 102, 180, 197, 199, and 200 and an antisense strand comprising a sequence complementary thereto.

4 . The siRNA of claim 1 , wherein the sense or antisense strand of the siRNA includes at least one chemical modification.

5 . The siRNA of claim 4 , wherein the chemical modification is at least one selected from modification by substitution of —OH group with —CH 3 (methyl), —OCH3 (methoxy), —NH2, —F (fluorine), —O-2-methoxyethyl, —O-propyl, —O-2-methylthioethyl, —O-3-aminopropyl, —O-3-dimethylaminopropyl, —O—N-methylacetamido, or —O-dimethylamidooxyethyl at a 2′-carbon site of a sugar structure in a nucleotide;
modification by substitution of oxygen in a sugar structure in the nucleotide with sulfur;
modification of a nucleotide bond into a phosphorothioate bond, a boranophosphate bond, or a methyl phosphonate bond; and
modification into a peptide nucleic acid (PNA) type, a locked nucleic acid (LNA) type, or a unlocked nucleic acid (UNA) type.

6 . The siRNA of claim 1 , wherein at least one phosphate group(s) is bound to a 5′-end of the antisense strand of the siRNA.

7 . Double-stranded oligo RNA molecule(s) comprising a structure of the following Structural Formula (1).
A-X—R—Y—B  Structural Formula (1)
where A is a hydrophilic compound, B is a hydrophobic compound, X and Y each are independently a simple covalent bond or a linker-mediated covalent bond, and R is Gankyrin or BMI-1 specific siRNA.

8 . The double-stranded oligo RNA molecule(s) of claim 7 , having a structure of Structural Formula (2)

where S is the sense strand of the siRNA of claim 7 , AS is the antisense strand thereof, and A, B, X, and Y have the same definitions as in claim 7 .

9 . The double-stranded oligo RNA molecule(s) of claim 8 , having a structure of Structural Formula (3)

where A, B, X, Y, S, and AS have the same definitions as those in claim 8 , and 5′ and 3′ mean a 5′-end and a 3′-end of the sense strand of the siRNA, respectively.

10 . The double-stranded oligo RNA molecule(s) of claim 7 , wherein the Gankyrin or BMI-1 specific siRNA comprises a sense strand comprising any one sequence selected from SEQ ID NOs. 1 to 200 and an antisense strand comprising a sequence complementary thereto.

11 . The double-stranded oligo RNA molecule(s) of claim 7 , wherein the hydrophilic compound has a molecular weight of 200 to 10,000.

12 . The double-stranded oligo RNA molecule(s) of claim 11 , wherein the hydrophilic compound is any one selected from a group consisting of polyethylene glycol (PEG), polyvinyl pyrrolidone, and polyoxazoline.

13 . The double-stranded oligo RNA molecule(s) of claim 7 , wherein the hydrophobic compound has a molecular weight of 250 to 1,000.

14 . The double-stranded oligo RNA molecule(s) of claim 13 , wherein the hydrophobic compound is any one selected from a group consisting of a steroid derivative, a glyceride derivative, glycerol ether, polypropylene glycol, saturated or unsaturated C12-C50 hydrocarbon, diacylphosphatidylcholine, fatty acid, phospholipid, and lipopolyamine.

15 . The double-stranded oligo RNA molecule(s) of claim 14 , wherein the steroid derivative is selected from a group consisting of cholesterol, cholestanol, cholic acid, cholesteryl formate, cholestanyl formate, and cholestearyl amine.

16 . The double-stranded oligo RNA molecule(s) of claim 14 , wherein the glyceride derivative is selected from mono-, di-, and tri-glycerides.

17 . The double-stranded oligo RNA molecule(s) of claim 7 , wherein the covalent bond represented by X and Y is a non-degradable bond or a degradable bond.

18 . The double-stranded oligo RNA molecule(s) of claim 17 , wherein the non-degradable bond is an amide bond or a phosphate bond.

19 . The double-stranded oligo RNA molecule(s) of claim 17 , wherein the degradable bond is a disulfide bond, an acid-degradable bond, an ester bond, an anhydride bond, a biodegradable bond, or an enzyme-degradable bond.

20 . The double-stranded oligo RNA molecule(s) of claim 7 , wherein a ligand which binds to a receptor promoting internalization into target cells through receptor-mediated endocytosis (REM) is additionally bound to the hydrophilic compound.

21 . The double-stranded oligo RNA molecule(s) of claim 20 , wherein the ligand is selected from a group consisting of a target receptor-specific antibody, aptamer, peptide, folate, N-acetyl galactosamine (NAG), glucose, and mannose.

22 . Nanoparticle(s) comprising the double-stranded oligo RNA molecule(s) of claim 7 .

23 . The nanoparticle(s) of claim 22 , composed by mixing double-stranded oligo RNA molecules containing siRNAs comprising different sequences with each other.
Gankyrin or BMI-1 specific siRNA comprising a sense strand comprising any one sequence selected from SEQ ID NOs. 1 to 200 and an antisense strand comprising a sequence complementary thereto

24 . A pharmaceutical composition comprising Gankyrin or BMI-1 specific siRNA comprising a sense strand comprising any one sequence selected from SEQ ID NOs. 1 to 200 and an antisense strand comprising a sequence complementary thereto, the double-stranded oligo RNA molecule(s) of claim 7 , or nanoparticle(s) comprising said double-stranded oligo RNA molecule(s) as an active ingredient.

25 . The pharmaceutical composition of claim 24 , wherein it is a pharmaceutical composition for preventing or treating cancer.

26 . The pharmaceutical composition of claim 25 , wherein cancer is selected from a group consisting of liver cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, breast cancer, ovarian cancer, kidney cancer, and lung cancer.

27 . The pharmaceutical composition of claim 26 , wherein liver cancer is hepatocellular carcinoma (HCC).

28 . Lypholized formulations comprising the pharmaceutical composition of claim 24 .

29 . A method for preventing or treating cancer characterized by administering Gankyrin or BMI-1 specific siRNA comprising a sense strand comprising any one sequence selected from SEQ ID NOs. 1 to 200 and an antisense strand comprising a sequence complementary thereto, the double-stranded oligo RNA molecule(s) of claim 7 , or the nanoparticle(s) comprising said double-stranded oligo RNA molecule(s), to an individual requiring such treatment or prevention of cancer.

30 . The method for preventing or treating cancer of claim 29 , wherein cancer is selected from a group consisting of liver cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, breast cancer, ovarian cancer, kidney cancer, and lung cancer.

31 . The method for preventing or treating cancer of claim 30 , wherein liver cancer is he

Assignees

• Bioneer Corp
• Sanofi Aventis Korea Co Ltd

Inventors

• Chae Jeiwook
• Park Han Oh
• Yoon Pyoung Oh
• Han Boram
• Kim Han-Na
• Yun Sung-Ii
• Park Jun Hong
• Ko Youngho
• Choi Gi-Eun
• Jung Junsoo
• Kim Jae Eun

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C12N15/1135 Primary

  against oncogenes or tumor suppressor genes · CPC title

• A61P1/16

  for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• A61K47/60

  the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol · CPC title