RNA interference agents for GST-PI gene modulation

Patent metadata

Field	Value
Publication number	US-10047111-B2
Application number	US-201514979574-A
Country	US
Kind code	B2
Filing date	Dec 28, 2015
Priority date	Dec 26, 2014
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

Abstract

This invention provides compounds, compositions and methods for modulating the expression of human GST-π using RNA interference. The RNA interference molecules can be used in methods for preventing or treating diseases such as malignant tumor. Provided are a range of siRNA structures, having one or more of nucleotides being modified or chemically-modified. Advantageous structures include siRNAs with 2′-deoxy nucleotides located in the seed region, as well as other nucleotide modifications.

First claim

What is claimed is:

1. A nucleic acid molecule for inhibiting expression of GST-π comprising a sense strand and an antisense strand, wherein the strands form a duplex region, and wherein the antisense strand is UAGGGUCUCAAAAGGCUUCNN SEQ ID NO:157 and the sense strand is GAAGCCUUUUGAGACCCUANN SEQ ID NO:131, wherein N is selected from the group of A, C, G, U, 2′-OMe-U, a, c, g, u, t, an inverted nucleotide, and a chemically modified nucleotide.

2. The nucleic acid molecule of claim 1 , wherein one or more of the nucleotides in the duplex region is chemically-modified.

3. The nucleic acid molecule of claim 2 , wherein the chemically-modified nucleotides are selected from 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, and any combination thereof.

4. The nucleic acid molecule of claim 2 , wherein the antisense strand has deoxynucleotides in a plurality of positions, the plurality of positions being one of the following:
each of positions 4, 6 and 8, from the 5′ end of the antisense strand;
each of positions 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 1, 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 3-8, from the 5′ end of the antisense strand; or
each of positions 5-8, from the 5′ end of the antisense strand.

5. The nucleic acid molecule of claim 4 , wherein the molecule has one or more 2′-deoxy-2′-fluoro substituted nucleotides in the duplex region.

6. The nucleic acid molecule of claim 2 , wherein the antisense strand is SEQ ID NO:182 and the sense strand is SEQ ID NO:156.

7. The nucleic acid molecule of claim 2 , wherein the antisense strand is SEQ ID NO:180 and the sense strand is SEQ ID NO:154.

8. The nucleic acid molecule of claim 2 , wherein the antisense strand is SEQ ID NO:181 and the sense strand is SEQ ID NO:155.

9. The nucleic acid molecule of claim 1 , wherein the molecule inhibits expression of GST-π mRNA in A549 cells with an IC50 of less than 50 pM.

10. A pharmaceutical composition comprising the nucleic acid molecule of claim 1 and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition of claim 10 , wherein the carrier comprises a lipid molecule or liposome.

12. A vector or cell comprising the nucleic acid molecule of claim 1 .

13. A method for treating pancreatic cancer or lung cancer, the method comprising administering to a subject in need a composition of claim 10 .

14. A nucleic acid molecule for inhibiting expression of GST-π comprising a sense strand and an antisense strand, wherein the strands form a duplex region, and wherein the antisense strand is ACAGCAGGGUCUCAAAAGGNN SEQ ID NO:195 and the sense strand is CCUUUUGAGACCCUGCUGUNN SEQ ID NO:183, wherein N is selected from the group of A, C, G, U, 2′-OMe-U, a, c, g, u, t, an inverted nucleotide, and a chemically modified nucleotide.

15. The nucleic acid molecule of claim 14 , wherein one or more of the nucleotides in the duplex region is chemically-modified.

16. The nucleic acid molecule of claim 15 , wherein the chemically-modified nucleotides are selected from 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, and any combination thereof.

17. The nucleic acid molecule of claim 15 , wherein the antisense strand has deoxynucleotides in a plurality of positions, which plurality of positions are one of the following:
each of positions 4, 6 and 8, from the 5′ end of the antisense strand;
each of positions 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 1, 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 3-8, from the 5′ end of the antisense strand; or
each of positions 5-8, from the 5′ end of the antisense strand.

18. The nucleic acid molecule of claim 17 , wherein the molecule has one or more 2′-deoxy-2′-fluoro substituted nucleotides in the duplex region.

19. The nucleic acid molecule of claim 14 , wherein the molecule inhibits expression of GST-π mRNA in A549 cells with an IC50 of less than 50 pM.

20. A pharmaceutical composition comprising the nucleic acid molecule of claim 14 and a pharmaceutically acceptable carrier.

21. The pharmaceutical composition of claim 20 , wherein the carrier is a lipid molecule or liposome.

22. A vector or cell comprising the nucleic acid molecule of claim 14 .

23. A method for treating pancreatic cancer or lung cancer, the method comprising administering to a subject in need a composition of claim 20 .

Assignees

• Nitto Denko Corp

Inventors

• Minomi Kenjirou
• Takahashi Hirokazu
• Terada Erika
• Harborth Jens
• Zhang Jun
• Ahmadian Mohammad
• Ying Wenbin

Classifications

• A61P35/00 Primary

  Antineoplastic agents · CPC title

• C07D311/30

  not hydrogenated in the hetero ring, e.g. flavones · CPC title

• C12Q1/02

  involving viable microorganisms · CPC title

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• A61K9/51

  Nanocapsules; {Nanoparticles; (nanotubes A61K9/0092; polymeric micelles A61K9/1075; polymersomes A61K9/1273; pure drug nanoparticles A61K9/14; drug nanoparticles with adsorbed surface modifiers A61K9/141; conjugates, e.g. between drug and non-active nanoparticles, A61K47/50; preparations for in vivo diagnosis A61K49/00; with radioactive substances A61K51/00)} · CPC title