Rna interference agents for gst-pi gene modulation

Patent metadata

Field	Value
Publication number	US-2016208265-A1
Application number	US-201514979574-A
Country	US
Kind code	A1
Filing date	Dec 28, 2015
Priority date	Dec 26, 2014
Publication date	Jul 21, 2016
Grant date	—

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 SEQ ID NOs:157 and the sense strand is SEQ ID NOs:131.

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

3 . The nucleic acid molecule of claim 2 , wherein the modified or chemically-modified nucleotides are 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, or 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 NOs:182 and the sense strand is SEQ ID NOs:156.

7 . The nucleic acid molecule of claim 2 , wherein the antisense strand is SEQ ID NOs:180 and the sense strand is SEQ ID NOs:154.

8 . The nucleic acid molecule of claim 2 , wherein the antisense strand is SEQ ID NOs:181 and the sense strand is SEQ ID NOs:155.

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

10 . The nucleic acid molecule of claim 1 , wherein a single administration of the molecule inhibits expression of GST-π mRNA levels by at least 25% in vivo.

11 . The nucleic acid molecule of claim 1 , wherein the passenger strand has off target activity reduced by at least 50-fold.

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

13 . The pharmaceutical composition of claim 12 , wherein the carrier comprises a lipid molecule or liposome.

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

15 . A method for treating a disease associated with GST-π expression, the method comprising administering to a subject in need a composition of claim 12 , wherein the disease is malignant tumor, cancer, cancer caused by cells expressing mutated KRAS, sarcoma, or carcinoma.

16 . 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 SEQ ID NOs:195 and the sense strand is SEQ ID NOs:183.

17 . The nucleic acid molecule of claim 16 , wherein one or more of the nucleotides in the duplex region is modified or chemically-modified.

18 . The nucleic acid molecule of claim 17 , wherein the modified or chemically-modified nucleotides are 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, or any combination thereof.

19 . The nucleic acid molecule of claim 17 , 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.

20 . The nucleic acid molecule of claim 19 , wherein the molecule has one or more 2′-deoxy-2′-fluoro substituted nucleotides in the duplex region.

21 . The nucleic acid molecule of claim 17 , wherein the antisense strand is SEQ ID NOs:205 and the sense strand is SEQ ID NOs:193.

22 . The nucleic acid molecule of claim 17 , wherein the antisense strand is SEQ ID NOs:202 and the sense strand is SEQ ID NOs:190.

23 . The nucleic acid molecule of claim 17 , wherein the antisense strand is SEQ ID NOs:199 and the sense strand is SEQ ID NOs:187.

24 . The nucleic acid molecule of claim 17 , wherein the antisense strand is SEQ ID NOs:201 and the sense strand is SEQ ID NOs:189.

25 . The nucleic acid molecule of claim 16 , wherein the molecule inhibits expression of GST-π mRNA with an IC50 of less than 50 pM.

26 . The nucleic acid molecule of claim 16 , wherein a single administration of the molecule inhibits expression of GST-π mRNA levels by at least 25% in vivo.

27 . The nucleic acid molecule of claim 16 , wherein the passenger strand has off target activity reduced by at least 50-fold.

28 . A pharmaceutical composition comprising the nucleic acid molecule of claim 16 and a pharmaceutically acceptable carrier.

29 . The pharmaceutical composition of claim 28 , wherein the carrier is a lipid molecule or liposome.

30 . A vector or cell comprising the nucleic acid molecule of claim 16 .

31 . A method for treating a disease associated with GST-π expression, the method comprising administering to a subject in need a composition of claim 28 , wherein the disease is malignant tumor, cancer, cancer caused by cells expressing mutated KRAS, sarcoma, or carcinoma.

32 . 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 SEQ ID NOs:222 and the sense strand is SEQ ID NOs:207.

33 . The nucleic acid molecule of claim 32 , wherein one or more of the nucleotides in the duplex region is modified or chemically-modified.

34 . The nucleic acid molecule of claim 33 , wherein the modified or chemically-modified nucleotides are 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, or any combination thereof.

35 . The nucleic acid molecule of claim 33 , 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.

36 . The nucleic acid molecule of claim 35 , wherein the molecule has one or more 2′-deoxy-2′-fluoro substituted nucleotides in the duplex region.

37 . The nucleic acid molecule of claim 33 , wherein the antisense strand is SEQ ID NOs:232 and the sense strand is SEQ ID NOs:217.

38 . T

Assignees

• Nitto Denko Corp

Inventors

• Niitsu Yoshiro
• Minomi Kenjirou
• Takahashi Hirokazu
• Terada Erika
• Harborth Jens
• Cina Cima
• Zhang Jun
• Ahmadian Mohammad
• Ying Wenbin

Classifications

• A61P35/00 Primary

  Antineoplastic agents · CPC title

• C12N15/1137 Primary

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• A61K31/713 Primary

  Double-stranded nucleic acids or oligonucleotides · CPC title

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• A61B5/0071 Primary

  by measuring fluorescence emission · CPC title