STAT6 siRNA

Abstract

The present disclosure relates to nucleic acid silencing molecules comprising or consisting of a small interfering RNA (siRNA) that reduces the expression of signal transduction and activator of transcription 6 (STAT6). The present disclosure also relates to nucleic acid silencing molecules comprising or consisting of a small interfering RNA (siRNA) that reduces the expression of signal transduction and activator of transcription 6 (STAT6). The disclosure also relates to an in vitro method of reducing STAT6 expression in a cell. The disclosure further relates to methods of treating a disease in a subject comprising administering to the subject a composition comprising the nucleic acid silencing molecule comprising or consisting of a siRNA that reduces the expression of STAT6, and related medical uses.

Description

Technical Field

[0001] This disclosure relates to nucleic acid silencing molecules comprising or composed of small interfering RNA (siRNA) that reduces the expression of signal transduction and activating factor 6 (STAT6). This disclosure also relates to in vitro methods for reducing STAT6 expression in cells. This disclosure further relates to methods for treating a disease in a subject and related medical uses, said methods comprising administering to the subject a composition comprising or composed of siRNA that reduces STAT6 expression. Background Technology

[0002] Signal transducers and activators of transcription (STATs) are a family of transcription factors containing SH2. STATs exist in an inactive form in the cytoplasm until they are activated by phosphorylation by Janus kinases (JAKs). JAKs are associated with the signal transduction chain of the thrombopoietin family of cytokine receptors. When these receptors bind cytokines, they dimerize. This allows JAKs on the signal transduction chain to cross-phosphorylate each other, thereby stimulating their kinase activity. They can then recruit STATs by phosphorylating conserved tyrosine residues at the C-terminus. This causes a conformational change in STATs, forming dimers that can exert downstream effects. In particular, phosphorylated STAT dimers enter the nucleus and act as transcription factors to initiate the expression of certain genes. Genes regulated by STATs include those related to the growth and differentiation of specific lymphocyte subsets.

[0003] Therefore, STATs are activated by cytokines, growth factors, and hormone signal transduction. There are seven types of STATs: STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT6. Typically, one cytokine activates one type of STAT. For example, IL-4 and IL-13 each activate STAT6. In turn, STAT6 initiates the expression of genes required for Th2 cell development.

[0004] Th2 helper cells provide T-cell assistance for humoral immune responses (i.e., antibody-mediated immunity). Their effector cytokines include IL-4, IL-5, IL-9, IL-10, IL-13, and IL-25, and their primary effector cells are eosinophils, basophils, mast cells, B cells, and IL-4 / IL-5 CD4 T cells. Th2 cells play a crucial role in immunity against extracellular parasites, bacteria, and allergens. However, overactivation of Th2 cells can lead to inflammatory and / or allergic diseases such as allergic rhinitis, atopic dermatitis, and asthma.

[0005] STAT6 is highly expressed in a variety of cancer types, including breast cancer, pancreatic cancer, endometrial cancer, colorectal cancer, skin cancer, lung cancer, prostate cancer, thyroid cancer, and colorectal cancer, and plays a crucial role in tumor initiation and malignant transformation. In cancer cells, STAT6 regulates the expression of genes involved in immune and inflammatory responses, cell survival, tumor proliferation, and metastasis. Cancers expressing STAT6 have been shown to be associated with lower survival rates, increased lymph node metastasis, and adverse alterations in inflammatory responses.

[0006] Therefore, the mechanism of reducing STAT6 expression is ideal for treating a range of diseases. Summary of the Invention

[0007] The inventors have identified numerous novel and beneficial small interfering RNAs (siRNAs) that reduce STAT6 expression. These newly identified STAT6 siRNAs can be used to treat diseases caused or contributing to adverse clinical effects by STAT6 expression. For example, the STAT6 siRNAs disclosed herein can be used to treat cancer or immune system diseases. For example, the STAT6 siRNAs disclosed herein can be used to treat inflammatory diseases.

[0008] Therefore, this disclosure provides a nucleic acid silencing molecule comprising or composed of a small interfering siRNA that reduces STAT6 expression, wherein the siRNA targets exons 5, 15, 19 or 10 of STAT6.

[0009] This disclosure further provides:

[0010] - An in vitro method for reducing STAT6 expression in cells, comprising: contacting the cells with a nucleic acid silencing molecule of the present disclosure;

[0011] - A method of treating a disease in a subject, comprising: administering to the subject a composition comprising a nucleic acid silencing molecule of the present disclosure;

[0012] - A composition comprising the nucleic acid silencing molecule disclosed herein, used in a method of treating a disease. Attached Figure Description

[0013] Figure 1 A) siRNA1, B) siRNA1w, C) siRNA1a, D) siRNA1b, E) siRNA1c and F) siRNA1d.

[0014] Figure 2 A) siRNA2, B) siRNA2a, C) siRNA2b, D) siRNA2c and E) siRNA2d.

[0015] Figure 3 A) siRNA3, B) siRNA3a, C) siRNA3b, D) siRNA3c and E) siRNA3d.

[0016] Figure 2 A) siRNA4, B) siRNA4a, C) siRNA4b, D) siRNA4c and E) siRNA4d.

[0017] Description of sequence listings

[0018] SEQ ID NO:1 – The base sequence of the positive strand of siRNA1 (i.e., the unmodified nucleotide sequence).

[0019] SEQ ID NO:2 – siRNA1w base sequence of the positive strand (i.e., the unmodified nucleotide sequence).

[0020] SEQ ID NO:3 – Base sequence of the positive strand of siRNA1a (i.e., the unmodified nucleotide sequence).

[0021] SEQ ID NO:4 – Base sequence of the positive strand of siRNA1b (i.e., the unmodified nucleotide sequence).

[0022] SEQ ID NO:5 – Base sequence of the positive strand of siRNA1c (i.e., the unmodified nucleotide sequence).

[0023] SEQ ID NO:6 – Base sequence of the positive strand of siRNA1d (i.e., the unmodified nucleotide sequence).

[0024] SEQ ID NO:7 – Base sequence of the positive strand of siRNA2 (i.e., the unmodified nucleotide sequence).

[0025] SEQ ID NO:8 – Base sequence of the positive strand of siRNA2a (i.e., the unmodified nucleotide sequence).

[0026] SEQ ID NO:9 – Base sequence of the positive strand of siRNA2b (i.e., the unmodified nucleotide sequence).

[0027] SEQ ID NO:10 – Base sequence of the positive strand of siRNA2c (i.e., the unmodified nucleotide sequence).

[0028] SEQ ID NO:11 – Base sequence of the positive strand of siRNA2d (i.e., the unmodified nucleotide sequence).

[0029] SEQ ID NO:12 – Base sequence of the positive strand of siRNA3 (i.e., the unmodified nucleotide sequence).

[0030] SEQ ID NO:13 – Base sequence of the positive strand of siRNA3a (i.e., the unmodified nucleotide sequence).

[0031] SEQ ID NO:14 – Base sequence of the positive strand of siRNA3b (i.e., the unmodified nucleotide sequence).

[0032] SEQ ID NO:15 – Base sequence of the positive strand of siRNA3c (i.e., the unmodified nucleotide sequence).

[0033] SEQ ID NO:16 – Base sequence of the positive strand of siRNA3d (i.e., the unmodified nucleotide sequence).

[0034] SEQ ID NO:17 – Base sequence of the positive strand of siRNA4 (i.e., the unmodified nucleotide sequence).

[0035] SEQ ID NO:18 – Base sequence of the positive strand of siRNA4a (i.e., the unmodified nucleotide sequence).

[0036] SEQ ID NO:19 – Base sequence of the positive strand of siRNA4b (i.e., the unmodified nucleotide sequence).

[0037] SEQ ID NO:20 – Base sequence of the positive strand of siRNA4c (i.e., the unmodified nucleotide sequence).

[0038] SEQ ID NO:21 – Base sequence of the positive strand of siRNA4d (i.e., the unmodified nucleotide sequence).

[0039] SEQ ID NO:22 – Preferred modified nucleotide sequence of the positive strand of siRNA1.

[0040] SEQ ID NO:23 – siRNA1w preferred modified nucleotide sequence of the positive strand.

[0041] SEQ ID NO:24 – Preferred modified nucleotide sequence of the positive strand of siRNA1a.

[0042] SEQ ID NO:25 – Preferred modified nucleotide sequence of the positive strand of siRNA1b.

[0043] SEQ ID NO:26 – Preferred modified nucleotide sequence of the positive strand of siRNA1c.

[0044] SEQ ID NO:27 – Preferred modified nucleotide sequence of the positive strand of siRNA1d.

[0045] SEQ ID NO:28 – Preferred modified nucleotide sequence of the positive strand of siRNA2.

[0046] SEQ ID NO:29 – Preferred modified nucleotide sequence of the positive strand of siRNA2a.

[0047] SEQ ID NO:30 – Preferred modified nucleotide sequence of the positive strand of siRNA2b.

[0048] SEQ ID NO:31 – Preferred modified nucleotide sequence of the positive strand of siRNA2c.

[0049] SEQ ID NO:32 – Preferred modified nucleotide sequence of the positive strand of siRNA2d.

[0050] SEQ ID NO:33 – Preferred modified nucleotide sequence of the positive strand of siRNA3.

[0051] SEQ ID NO:34 – Preferred modified nucleotide sequence of the positive strand of siRNA3a.

[0052] SEQ ID NO:35 – Preferred modified nucleotide sequence of the positive strand of siRNA3b.

[0053] SEQ ID NO:36 – Preferred modified nucleotide sequence of the positive strand of siRNA3c.

[0054] SEQ ID NO:37 – Preferred modified nucleotide sequence of the positive strand of siRNA3d.

[0055] SEQ ID NO:38 – Preferred modified nucleotide sequence of the positive strand of siRNA4.

[0056] SEQ ID NO:39 – Preferred modified nucleotide sequence of the positive strand of siRNA4a.

[0057] SEQ ID NO:40 – Preferred modified nucleotide sequence of the positive strand of siRNA4b.

[0058] SEQ ID NO:41 – Preferred modified nucleotide sequence of the positive strand of siRNA4c.

[0059] SEQ ID NO:42 – Preferred modified nucleotide sequence of the positive strand of siRNA4d.

[0060] SEQ ID NO:43 – The base sequence of the antisense strand of siRNA1 (i.e., the unmodified nucleotide sequence).

[0061] SEQ ID NO:44 – The base sequence of the antisense strand of siRNA1w (i.e., the unmodified nucleotide sequence).

[0062] SEQ ID NO:45 – Base sequence of the antisense strand of siRNA1a (i.e., the unmodified nucleotide sequence).

[0063] SEQ ID NO:46 – The base sequence of the antisense strand of siRNA1b (i.e., the unmodified nucleotide sequence).

[0064] SEQ ID NO:47 – The base sequence of the antisense strand of siRNA1c (i.e., the unmodified nucleotide sequence).

[0065] SEQ ID NO:48 – The base sequence of the antisense strand of siRNA1d (i.e., the unmodified nucleotide sequence).

[0066] SEQ ID NO:49 – The base sequence of the antisense strand of siRNA2 (i.e., the unmodified nucleotide sequence).

[0067] SEQ ID NO:50 – The base sequence of the antisense strand of siRNA2a (i.e., the unmodified nucleotide sequence).

[0068] SEQ ID NO:51 – Base sequence of the antisense strand of siRNA2b (i.e., the unmodified nucleotide sequence).

[0069] SEQ ID NO:52 – The base sequence of the antisense strand of siRNA2c (i.e., the unmodified nucleotide sequence).

[0070] SEQ ID NO:53 – The base sequence of the antisense strand of siRNA2d (i.e., the unmodified nucleotide sequence).

[0071] SEQ ID NO:54 – The base sequence of the antisense strand of siRNA3 (i.e., the unmodified nucleotide sequence).

[0072] SEQ ID NO:55 – Base sequence of the antisense strand of siRNA3a (i.e., the unmodified nucleotide sequence).

[0073] SEQ ID NO:56 – Base sequence of the antisense strand of siRNA3b (i.e., the unmodified nucleotide sequence).

[0074] SEQ ID NO:57 – The base sequence of the antisense strand of siRNA3c (i.e., the unmodified nucleotide sequence).

[0075] SEQ ID NO:58 – The base sequence of the antisense strand of siRNA3d (i.e., the unmodified nucleotide sequence).

[0076] SEQ ID NO:59 – The base sequence of the antisense strand of siRNA4 (i.e., the unmodified nucleotide sequence).

[0077] SEQ ID NO:60 – The base sequence of the antisense strand of siRNA4a (i.e., the unmodified nucleotide sequence).

[0078] SEQ ID NO:61 – Base sequence of the antisense strand of siRNA4b (i.e., the unmodified nucleotide sequence).

[0079] SEQ ID NO:62 – Base sequence of the antisense strand of siRNA4c (i.e., the unmodified nucleotide sequence).

[0080] SEQ ID NO:63 – Base sequence of the antisense strand of siRNA4d (i.e., the unmodified nucleotide sequence).

[0081] SEQ ID NO:64 – Preferred modified nucleotide sequence of the antisense strand of siRNA1.

[0082] SEQ ID NO:65 – Preferred modified nucleotide sequence of the antisense strand of siRNA1w.

[0083] SEQ ID NO:66 – Preferred modified nucleotide sequence of the antisense strand of siRNA1a.

[0084] SEQ ID NO:67 – Preferred modified nucleotide sequence of the antisense strand of siRNA1b.

[0085] SEQ ID NO:68 – Preferred modified nucleotide sequence of the siRNA1c antisense strand.

[0086] SEQ ID NO:69 – Preferred modified nucleotide sequence of the antisense strand of siRNA1d.

[0087] SEQ ID NO:70 – Preferred modified nucleotide sequence of the antisense strand of siRNA2.

[0088] SEQ ID NO:71 – Preferred modified nucleotide sequence of the antisense strand of siRNA2a.

[0089] SEQ ID NO:72 – Preferred modified nucleotide sequence of the antisense strand of siRNA2b.

[0090] SEQ ID NO:73 – Preferred modified nucleotide sequence of the siRNA2c antisense strand.

[0091] SEQ ID NO:74 – Preferred modified nucleotide sequence of the siRNA2d antisense strand.

[0092] SEQ ID NO:75 – Preferred modified nucleotide sequence of the antisense strand of siRNA3.

[0093] SEQ ID NO:76 – Preferred modified nucleotide sequence of the antisense strand of siRNA3a.

[0094] SEQ ID NO:77 – Preferred modified nucleotide sequence of the antisense strand of siRNA3b.

[0095] SEQ ID NO:78 – Preferred modified nucleotide sequence of the siRNA3c antisense strand.

[0096] SEQ ID NO:79 – Preferred modified nucleotide sequence of the antisense strand of siRNA3d.

[0097] SEQ ID NO:80 – Preferred modified nucleotide sequence of the antisense strand of siRNA4.

[0098] SEQ ID NO:81 – Preferred modified nucleotide sequence of the antisense strand of siRNA4a.

[0099] SEQ ID NO:82 – Preferred modified nucleotide sequence of the antisense strand of siRNA4b.

[0100] SEQ ID NO:83 – Preferred modified nucleotide sequence of the siRNA4c antisense strand.

[0101] SEQ ID NO:84 – Preferred modified nucleotide sequence of the antisense strand of siRNA4d. Detailed Implementation

[0102] It should be understood that different applications of the disclosed methods and products can be customized to meet specific needs in the field. It should also be understood that the terminology used herein is for describing specific embodiments of this disclosure only and is not intended to be limiting.

[0103] All publications, patents, and patent applications cited in this article, whether above or below, are incorporated herein by reference in their entirety.

[0104] General definition

[0105] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0106] Unless otherwise expressly specified, the singular forms “a”, “an”, and “the” as used in this specification and the appended claims include plural references. Thus, for example, reference to “a siRNA” includes “multiple siRNAs,” and reference to “antigen” includes two or more such antigens, etc.

[0107] Generally, the term "contains" is intended to mean, but is not limited to, siRNA. For example, the phrase "nucleic acid silencing molecule containing siRNA" should be interpreted as meaning that the nucleic acid silencing molecule contains siRNA, but the nucleic acid silencing molecule may contain additional nucleotides or sequences.

[0108] In some aspects of this disclosure, the word "comprising" is replaced by the phrase "composed of". The term "composed of" is intended to be restrictive. For example, the phrase "nucleic acid silencing molecule composed of siRNA" should be understood to mean that the nucleic acid silencing molecule contains siRNA and has no additional nucleotides or sequences.

[0109] In this article, the terms “protein” and “polypeptide” are used interchangeably and are intended to refer to amino acid polymer chains of any length.

[0110] For the purposes of this disclosure, in order to determine the percentage of identity between two sequences (e.g., two polynucleotide or two polypeptide sequences), the sequences are aligned for optimal comparison purposes (e.g., a gap may be introduced in the first sequence for optimal alignment with the second sequence). Nucleotide residues at nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, the nucleotides are identical at that position. The percentage of identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., identity % = number of identical positions / total number of positions in the reference sequence x 100).

[0111] Typically, sequence comparisons are performed over the entire length of a reference sequence. For example, if a user wants to determine whether a given (“test”) sequence shares a certain percentage of identity with SEQ ID NO:X, then SEQ ID NO:X would be the reference sequence. For instance, to assess whether a sequence shares at least 80% identity with SEQ ID NO:X (an instance of the reference sequence), a technician would perform a comparison over the length of SEQ ID NO:X and identify how many positions in the test sequence are identical to those in SEQ ID NO:X. If at least 80% of the positions are identical, then the test sequence shares at least 80% identity with SEQ ID NO:X. If the sequence is shorter than SEQ ID NO:X, empty or missing positions should be considered as dissimilar positions.

[0112] Those skilled in the art will recognize the various computer programs that can be used to determine homology or identity between two sequences. For example, mathematical algorithms can be used to compare sequences and determine the percentage of identity between two sequences.

[0113] Nucleic acid silencing molecules

[0114] This article discloses a nucleic acid silencing molecule that reduces STAT6 expression.

[0115] These silencing molecules can be used therapeutically to reduce STAT6 expression. As mentioned above, STAT6 is highly expressed in a variety of cancers, where it regulates the expression of genes involved in immune responses, cell survival, tumor proliferation, and metastasis. Therefore, reducing STAT6 expression in cancer cells can kill cancer cells, reduce or prevent tumor growth, and reduce or prevent metastasis. Furthermore, it has been demonstrated that STAT6 expression by immune cells in the tumor microenvironment contributes to the formation of an immunosuppressive microenvironment conducive to tumor maintenance and growth. Reducing STAT6 expression in the microenvironment can promote the formation of a pro-inflammatory microenvironment conducive to killing tumor cells.

[0116] For example, reducing STAT6 expression may be beneficial in diseases caused by or associated with an overactive Th2 response. Th2 responses are associated with inflammatory diseases, allergic diseases, graft-versus-host disease (GVHD), and autoimmune diseases. Since STAT6 is a key signaling transduction molecule for inducing and maintaining Th2 responses, reducing STAT6 expression in subjects can decrease the quantity and / or activity of Th2 responses. This can thus reduce or eliminate clinical signs or symptoms of an overactive Th2 response and may halt the progression of clinical signs or symptoms.

[0117] Other uses for the nucleic acid silencing molecules disclosed in this paper can also be envisioned. For example, nucleic acid silencing molecules can be used to experimentally reduce STAT6 expression in vivo or in vitro. In other words, nucleic acid silencing molecules can be used as research tools, such as for studying cancer, inflammatory diseases, allergic diseases, or autoimmune diseases.

[0118] Reduce STAT6 expression

[0119] This article discloses nucleic acid silencing molecules that reduce STAT6 expression. For example, nucleic acid silencing molecules can reduce STAT6 mRNA expression. For example, nucleic acid silencing molecules can reduce STAT6 protein expression. For example, nucleic acid silencing molecules can reduce the expression of both STAT6 mRNA and STAT6 protein. For example, the reduction in STAT6 mRNA expression can lead to a decrease in STAT6 protein expression.

[0120] For example, nucleic acid silencing molecules can reduce STAT6 expression in cells. For example, the cells can be human cells or cells derived from human cell lines. For example, the cells can be in vivo, in vitro, or ex vivo. The cells can be cells that have come into contact with nucleic acid silencing molecules. For example, reduced STAT6 expression in cells can refer to a decrease in the amount of STAT6 mRNA and / or STAT6 protein contained in the cells. Therefore, cells with reduced STAT6 expression can, for example, contain reduced amounts of STAT6 mRNA. Cells with reduced STAT6 expression can, for example, contain reduced amounts of STAT6 protein. Cells with reduced STAT6 expression can, for example, contain reduced amounts of STAT6 mRNA and reduced amounts of STAT6 protein.

[0121] For example, compared to cells that have not been exposed to the nucleic acid silencing molecule, the nucleic acid silencing molecule can reduce the expression of STAT6 in cells that have been exposed to it. For example, the cells exposed to the nucleic acid silencing molecule can be human cells or cells derived from human cell lines. For example, the cells exposed to the nucleic acid silencing molecule can be in vivo, in vitro, or ex vivo. For example, the cells that have not been exposed to the nucleic acid silencing molecule can be human cells or cells derived from human cell lines. For example, the cells that have not been exposed to the nucleic acid silencing molecule can be in vivo, in vitro, or ex vivo. Preferably, the cells exposed to the nucleic acid silencing molecule and the cells that have not been exposed to the nucleic acid silencing molecule are the same type of cells (e.g., human cells or cells derived from human cell lines). Preferably, the cells exposed to the nucleic acid silencing molecule and the cells that have not been exposed to the nucleic acid silencing molecule are under the same conditions (e.g., in vivo, in vitro, or ex vivo). Cells that have not been exposed to the nucleic acid silencing molecule may instead be exposed to a control molecule (e.g., a mimic nucleic acid silencing molecule). Mimic nucleic acid silencing molecules (e.g., mimic siRNA) and their design and production methods are well known in the art. For example, decreased STAT6 expression in cells exposed to nucleic acid silencing molecules can refer to a reduction in the amount of STAT6 mRNA and / or STAT6 protein present in these cells compared to cells not exposed to the silencing molecules. Therefore, cells exposed to nucleic acid silencing molecules and exhibiting decreased STAT6 expression may contain a reduced amount of STAT6 mRNA compared to cells not exposed to the silencing molecules. Cells exposed to nucleic acid silencing molecules and exhibiting decreased STAT6 expression may also contain a reduced amount of STAT6 protein compared to cells not exposed to the silencing molecules. Furthermore, cells exposed to nucleic acid silencing molecules and exhibiting decreased STAT6 expression may contain both reduced amounts of STAT6 mRNA and reduced amounts of STAT6 protein compared to cells not exposed to the silencing molecules.

[0122] In any of the foregoing aspects, for example, a nucleic acid silencing molecule can reduce STAT6 expression by at least 30%. In other words, a nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein by at least 30%. For example, a nucleic acid silencing molecule can reduce STAT6 expression by at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99%. In other words, for example, a nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein by at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99%. For example, nucleic acid silencing molecules can reduce STAT6 expression by 30% to 99%, such as 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%. That is, for example, nucleic acid silencing molecules can reduce the amount of STAT6 mRNA and / or the amount of STAT6 protein by 30% to 99%, such as 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%.

[0123] Preferably, the nucleic acid silencing molecule reduces STAT6 expression in cells by at least 30%. In other words, the nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein in cells by at least 30%. The cell types and conditions used to assess STAT6 expression are as described above. For example, the nucleic acid silencing molecule can reduce STAT6 expression in cells by at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99%. In other words, for example, the nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein in cells by at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99%. For example, nucleic acid silencing molecules can reduce STAT6 expression in cells by 30% to 99%, such as 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%. That is, for example, nucleic acid silencing molecules can reduce the amount of STAT6 mRNA and / or STAT6 protein in cells by 30% to 99%, such as 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%.

[0124] More preferably, the nucleic acid silencing molecule reduces STAT6 expression by at least 30% in cells that have not been exposed to it, relative to cells that have not been exposed to it. In other words, the nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein by at least 30% in cells that have been exposed to it, relative to cells that have not been exposed to it. The cell types and conditions used to assess STAT6 expression are as described above. For example, the nucleic acid silencing molecule can reduce STAT6 expression by at least 30%, at least 35%, at least 40%, 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% in cells that have been exposed to it, relative to cells that have not been exposed to it. In other words, for example, relative to cells that have not been exposed to the nucleic acid silencing molecule, the nucleic acid silencing molecule can reduce the amount of STAT6 mRNA and / or STAT6 protein in cells that have been exposed to the nucleic acid silencing molecule by at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99%. For example, relative to cells that have not been exposed to the nucleic acid silencing molecule, the nucleic acid silencing molecule can reduce STAT6 expression in cells that have been exposed to the nucleic acid silencing molecule by 30% to 99%, such as 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%. That is, for example, relative to cells that have not been exposed to nucleic acid silencing molecules, nucleic acid silencing molecules can reduce the amount of STAT6 mRNA and / or STAT6 protein in cells that have been exposed to nucleic acid silencing molecules by 30% to 99%, for example 35% to 95%, 40% to 90%, 45% to 85%, 50% to 80%, 55% to 75%, or 60% to 70%.

[0125] In any of the above aspects, nucleic acid silencing molecules can reduce STAT6 expression by 100%. In other words, nucleic acid silencing molecules can completely eliminate STAT6 expression. For example, nucleic acid silencing molecules can completely eliminate STAT6 mRNA expression (i.e., reduce STAT6 mRNA expression by 100%). For example, nucleic acid silencing molecules can completely eliminate STAT6 protein expression (i.e., reduce STAT6 protein expression by 100%). For example, nucleic acid silencing molecules can completely eliminate both STAT6 mRNA and STAT6 protein expression (i.e., reduce both STAT6 mRNA and STAT6 protein expression by 100%).

[0126] For example, nucleic acid silencing molecules can completely eliminate STAT6 expression in cells. Therefore, the cells may not express STAT6 mRNA and / or STAT6 protein. For example, the cells may be human cells or cells derived from human cell lines. For example, the cells may be in vivo, in vitro, or ex vivo. The cells may be cells that have been in contact with nucleic acid silencing molecules.

[0127] For example, compared to cells that have not been exposed to the nucleic acid silencing molecule, the nucleic acid silencing molecule can reduce STAT6 expression by 100% in cells that have been exposed to it. Cells exposed to the nucleic acid silencing molecule can be, for example, human cells or cells derived from human cell lines. Cells exposed to the nucleic acid silencing molecule can be, for example, in vivo, in vitro, or ex vivo. Cells not exposed to the nucleic acid silencing molecule can be, for example, human cells or cells derived from human cell lines. Cells not exposed to the nucleic acid silencing molecule can be, for example, in vivo, in vitro, or ex vivo. Preferably, the cells exposed to the nucleic acid silencing molecule and the cells not exposed to the nucleic acid silencing molecule are the same type of cell (e.g., human cells or cells derived from human cell lines). Preferably, the cells exposed to the nucleic acid silencing molecule and the cells not exposed to the nucleic acid silencing molecule are under the same conditions (e.g., in vivo, in vitro, or ex vivo). Cells not exposed to the nucleic acid silencing molecule may instead be exposed to a control molecule (e.g., a mimic nucleic acid silencing molecule). As described above, mimic nucleic acid silencing molecules (e.g., mimic siRNA) and their design and production methods are well known in the art.

[0128] As described above, where nucleic acid silencing molecules can, for example, reduce STAT6 expression in cells that have been exposed to nucleic acid silencing molecules relative to cells that have not been exposed to them, IL-4 expression in cells can also be reduced by a corresponding percentage relative to cells that have not been exposed to nucleic acid silencing molecules.

[0129] Nucleic acid silencing molecules

[0130] In the context of this disclosure, a silencing molecule can be defined as a molecule that reduces or eliminates (i.e., knocks down) the expression of a target gene. For example, a silencing molecule can reduce the amount of the mRNA product of a target gene. For example, a silencing molecule can eliminate the mRNA product of a target gene. For example, a silencing molecule can reduce the amount of the protein product of a target gene. For example, a silencing molecule can eliminate the protein product of a target gene. In this disclosure, the target gene is STAT6.

[0131] In the context of this disclosure, a nucleic acid silencing molecule can be defined as a silencing molecule containing one or more nucleic acids or composed of one or more nucleic acids. The nucleic acid silencing molecule of this disclosure may contain RNA. The nucleic acid silencing molecule of this disclosure may contain DNA. The nucleic acid silencing molecule of this disclosure may contain both DNA and RNA. The nucleic acid silencing molecule of this disclosure may be composed of RNA. The nucleic acid silencing molecule of this disclosure may be composed of both DNA and RNA.

[0132] Nucleic acid silencing molecules can reduce or eliminate (i.e., knock down) STAT6 expression through any mechanism known in the art. Since nucleic acid silencing molecules contain siRNA, they can reduce or eliminate STAT6 expression through known siRNA mechanisms. For example, nucleic acid silencing molecules can bind to the mRNA molecule encoded by the STAT6 gene to induce mRNA degradation (e.g., enzymatic degradation).

[0133] The length of a nucleic acid silencing molecule can be from about 10 to about 100 nucleotides, for example, from about 15 to about 95, about 20 to about 90, about 25 to about 85, about 30 to about 80, about 40 to about 75, about 45 to about 70, about 50 to about 65, or about 55 to about 60 nucleotides. Preferably, the length of the nucleic acid silencing molecule is less than 50 nucleotides (e.g., less than 45, less than 40, less than 35, or less than 30). The length of a nucleic acid silencing molecule can be, for example, from about 15 to about 30 nucleotides. For example, the length of a nucleic acid silencing molecule can be from about 16 to about 29, about 17 to about 28, about 18 to about 27, about 19 to about 26, or about 20 to about 25 nucleotides. Preferably, the length of the nucleic acid molecule is from about 20 to about 25 nucleotides. The length of a nucleic acid molecule can be, for example, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, or about 30 nucleotides. The preferred length of a nucleic acid molecule is about 25 nucleotides. Typical lengths of small interfering RNA (siRNA) are well known in the art.

[0134] For example, a nucleic acid silencing molecule may comprise one or more 2'-O-methyl (2'-OMe) modified nucleotides or be composed of 2'-O-methyl (2'-OMe) modified nucleotides. For example, a nucleic acid silencing molecule may comprise one or more 2'-fluorine (2'-F) modified nucleotides or be composed of 2'-fluorine (2'-F) modified nucleotides. For example, a nucleic acid silencing molecule may comprise one or more nucleotide thiophosphates or be composed of nucleotide thiophosphates. 2'-OMe modified nucleotides, 2'-F modified nucleotides, and nucleotide thiophosphates have been described in the art.

[0135] The siRNA targets STAT6. Therefore, the nucleic acid silencing molecule may be able to bind to the RNA encoded by the STAT6 gene. The nucleic acid silencing molecule may be able to bind to a portion of the RNA encoded by the STAT6 gene. For example, binding can be achieved through hybridization.

[0136] Nucleic acid silencing molecules can target RNA encoded by the STAT6 gene. For example, nucleic acid silencing molecules can target mRNA encoded by the STAT6 gene. Nucleic acid silencing molecules that "target" a specific nucleic acid sequence can bind to (e.g., hybridize to) that nucleic acid sequence.

[0137] Specifically, the siRNA targets exons 5, 15, 19, or 10 of STAT6. Therefore, the siRNA can bind to sequences contained in exons 5, 15, 19, or 10 of mRNA transcribed from STAT6. The siRNA can hybridize to sequences contained in exons 5, 15, 19, or 10 of mRNA transcribed from STAT6, for example, via Watson-Crick base pairing. One strand of the siRNA can bind to sequences contained in exons 5, 15, 19, or 10 of mRNA transcribed from STAT6. One strand of the siRNA can hybridize to sequences contained in exons 5, 15, 19, or 10 of mRNA transcribed from STAT6, for example, via Watson-Crick base pairing.

[0138] In the embodiments, siRNAs 1, 1w, 1a, 1b, 1c, and 1d target exon 5 of STAT6; siRNAs 2, 2a, 2b, 2c, and 2d target exon 15 of STAT6; siRNAs 3, 3a, 3b, 3c, and 3d target exon 19 of STAT6; and siRNAs 4, 4a, 4b, 4c, and 4d target exon 10 of STAT6. Therefore, siRNAs containing one of SEQ ID NOs: 1 to 6 or 43 to 48 target exon 5 of STAT6. siRNAs containing one of SEQ ID NOs: 7 to 11 or 49 to 53 target exon 15 of STAT6. siRNAs containing one of SEQ ID NOs: 12 to 16 or 54 to 58 target exon 19 of STAT6. siRNAs containing one of SEQ ID NOs: 17 to 21 or 59 to 63 target exon 10 of STAT6.

[0139] For example, a nucleic acid silencing molecule may comprise or consist of siRNA that reduces STAT6 expression, wherein the positive strand of the siRNA comprises any one of SEQ ID NO: 1 to 21 or has at least 75% sequence identity with it.

[0140] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:1 to 21 may, for example, contain at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any one of SEQ ID NO:1 to 21. Therefore, the positive strand of the siRNA may contain a nucleotide sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any one of SEQ ID NO:1 to 21.

[0141] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:1 to 21 may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide substitutions (e.g., two or more, three or more, four or more, or five or more) relative to one of SEQ ID NO:1 to 21. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:1 to 21 may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide deletions relative to one of SEQ ID NO:1 to 21. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:1 to 21 may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide insertions relative to one of SEQ ID NO:1 to 21.

[0142] Typically, siRNA contained in nucleic acid silencing molecules further includes an antisense strand. The antisense strand is able to hybridize to the sense strand via Watson-Crick base pairing.

[0143] Nucleic acid molecules may, for example, contain siRNA, wherein:

[0144] (1) The sense strand of the siRNA contains SEQ ID NO:3 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:45 or a nucleotide sequence having at least 75% sequence identity with it;

[0145] (2) The sense strand of the siRNA contains SEQ ID NO:4 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:46 or a nucleotide sequence having at least 75% sequence identity with it;

[0146] (3) The sense strand of the siRNA contains SEQ ID NO:5 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:47 or a nucleotide sequence having at least 75% sequence identity with it;

[0147] (4) The sense strand of the siRNA contains SEQ ID NO:6 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:48 or a nucleotide sequence having at least 75% sequence identity with it;

[0148] (5) The sense strand of the siRNA contains SEQ ID NO:1 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:43 or a nucleotide sequence having at least 75% sequence identity with it;

[0149] (6) The sense strand of the siRNA contains SEQ ID NO:2 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:44 or a nucleotide sequence having at least 75% sequence identity with it;

[0150] (7) The sense strand of the siRNA contains SEQ ID NO:7 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:49 or a nucleotide sequence having at least 75% sequence identity with it;

[0151] (8) The sense strand of the siRNA contains SEQ ID NO:8 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:50 or a nucleotide sequence having at least 75% sequence identity with it;

[0152] (9) The sense strand of the siRNA contains SEQ ID NO:9 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:51 or a nucleotide sequence having at least 75% sequence identity with it;

[0153] (10) The sense strand of the siRNA contains SEQ ID NO:10 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:52 or a nucleotide sequence having at least 75% sequence identity with it;

[0154] (11) The sense strand of the siRNA contains SEQ ID NO:11 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:53 or a nucleotide sequence having at least 75% sequence identity with it;

[0155] (12) The sense strand of the siRNA contains SEQ ID NO:12 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:54 or a nucleotide sequence having at least 75% sequence identity with it;

[0156] (13) The sense strand of the siRNA contains SEQ ID NO:13 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:55 or a nucleotide sequence having at least 75% sequence identity with it;

[0157] (14) The sense strand of the siRNA contains SEQ ID NO:14 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:56 or a nucleotide sequence having at least 75% sequence identity with it;

[0158] (15) The sense strand of the siRNA contains SEQ ID NO:15 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:57 or a nucleotide sequence having at least 75% sequence identity with it;

[0159] (16) The sense strand of the siRNA contains SEQ ID NO:16 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:58 or a nucleotide sequence having at least 75% sequence identity with it;

[0160] (17) The sense strand of the siRNA contains SEQ ID NO:17 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:59 or a nucleotide sequence having at least 75% sequence identity with it;

[0161] (18) The sense strand of the siRNA contains SEQ ID NO:18 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:60 or a nucleotide sequence having at least 75% sequence identity with it;

[0162] (19) The sense strand of the siRNA contains SEQ ID NO:19 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:61 or a nucleotide sequence having at least 75% sequence identity with it;

[0163] (20) The sense strand of the siRNA comprises SEQ ID NO:20 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA comprises SEQ ID NO:62 or a nucleotide sequence having at least 75% sequence identity with it; or

[0164] (21) The sense strand of the siRNA contains SEQ ID NO:21 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:63 or a nucleotide sequence having at least 75% sequence identity with it.

[0165] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:43 to 63 may, for example, contain a sequence identity of at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% with any one of SEQ ID NO:1 to 21. Therefore, the antisense strand of the siRNA may contain a nucleotide sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with any one of SEQ ID NO:43 to 63.

[0166] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:43 to 63 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide substitutions (e.g., two or more, three or more, four or more, or five or more) relative to one of SEQ ID NO:43 to 63. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:43 to 63 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide deletions relative to one of SEQ ID NO:43 to 63. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:43 to 63 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide insertions relative to one of SEQ ID NO:43 to 63.

[0167] Any one of SEQ ID NO:1 to 21 and 43 to 64 may contain one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, or nineteen or more) nucleotides modified with 2'-O-methyl (2'-OMe), or may be composed of nucleotides modified with 2'-O-methyl (2'-OMe). Nucleotides modified with 2'-O-methyl (2'-OMe) may be present at any position within SEQ ID NO:1 to 21 and 43 to 63.

[0168] Any one of SEQ ID NO:1 to 21 and 43 to 64 may contain one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, or nineteen or more) 2'-fluoro(2'-F) modified nucleotides, or may be composed of 2'-fluoro(2'-F) modified nucleotides. 2'-F modified nucleotides may be present at any position within SEQ ID NO:1 to 21 and 43 to 63.

[0169] Any one of SEQ ID NO:1 to 21 and 43 to 63 may contain one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, or nineteen or more) nucleotide thiophosphates or be composed of nucleotide thiophosphates. Nucleotide thiophosphates may be present at any position within SEQ ID NO:1 to 21 and 43 to 63.

[0170] Any one of SEQ ID NO: 11 to 21 and 43 to 63 may contain any combination of the following: (i) one or more 2'-OMe modified nucleotides, (ii) one or more 2'-F modified nucleotides, and (iii) one or more nucleotide thiophosphates. For example, any one of SEQ ID NO: 1 to 21 and 43 to 63 may contain (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii).

[0171] For illustrative purposes, exemplary modifications of SEQ ID NO:1 to 21 and 43 to 63 are shown. Figures 1 to 4 Exemplary modifications include 2'-OMe, 2'-F, and phosphate thioester modifications. The exemplified modifications are not limiting. Any type of chemical modification (e.g., 2'-OMe modification, 2'-F modification, nucleotide phosphate thioester modification) can be performed at any or all of the exemplified positions. Chemical modifications (e.g., 2'-OMe modification, 2'-F modification, nucleotide phosphate thioester modification) can also be performed at non-exemplified positions.

[0172] For example, the positive strand of the siRNA may contain any one of SEQ ID NO:22 to 42 or a nucleotide sequence having at least 75% sequence identity with it. The nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:22 to 42 may, for example, contain at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any one of SEQ ID NO:1 to 21. Therefore, the positive strand of the siRNA may contain a nucleotide sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with one of SEQ ID NO:22 to 42. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:22 to 42 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide substitutions (e.g., two or more, three or more, four or more, or five or more) relative to one of SEQ ID NO:22 to 42. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:22 to 42 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide deletions relative to one of SEQ ID NO:22 to 42. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:22 to 42 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide insertions relative to one of SEQ ID NO:22 to 42.

[0173] For example, nucleic acid molecules can contain siRNA, in which:

[0174] (1) The sense strand of the siRNA contains SEQ ID NO:24 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:66 or a nucleotide sequence having at least 75% sequence identity with it;

[0175] (2) The sense strand of the siRNA contains SEQ ID NO:25 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:67 or a nucleotide sequence having at least 75% sequence identity with it;

[0176] (3) The sense strand of the siRNA contains SEQ ID NO:26 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:68 or a nucleotide sequence having at least 75% sequence identity with it;

[0177] (4) The sense strand of the siRNA contains SEQ ID NO:27 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:69 or a nucleotide sequence having at least 75% sequence identity with it;

[0178] (5) The sense strand of the siRNA contains SEQ ID NO:22 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:64 or a nucleotide sequence having at least 75% sequence identity with it;

[0179] (6) The sense strand of the siRNA contains SEQ ID NO:23 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:65 or a nucleotide sequence having at least 75% sequence identity with it;

[0180] (7) The sense strand of the siRNA contains SEQ ID NO:28 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:70 or a nucleotide sequence having at least 75% sequence identity with it;

[0181] (8) The sense strand of the siRNA contains SEQ ID NO:29 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:71 or a nucleotide sequence having at least 75% sequence identity with it;

[0182] (9) The sense strand of the siRNA contains SEQ ID NO:30 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:72 or a nucleotide sequence having at least 75% sequence identity with it;

[0183] (10) The sense strand of the siRNA contains SEQ ID NO:31 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:73 or a nucleotide sequence having at least 75% sequence identity with it;

[0184] (11) The sense strand of the siRNA contains SEQ ID NO:32 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:74 or a nucleotide sequence having at least 75% sequence identity with it;

[0185] (12) The sense strand of the siRNA contains SEQ ID NO:33 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:75 or a nucleotide sequence having at least 75% sequence identity with it;

[0186] (13) The sense strand of the siRNA contains SEQ ID NO:34 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:76 or a nucleotide sequence having at least 75% sequence identity with it;

[0187] (14) The sense strand of the siRNA contains SEQ ID NO:35 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:77 or a nucleotide sequence having at least 75% sequence identity with it;

[0188] (15) The sense strand of the siRNA contains SEQ ID NO:36 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:78 or a nucleotide sequence having at least 75% sequence identity with it;

[0189] (16) The sense strand of the siRNA contains SEQ ID NO:37 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:79 or a nucleotide sequence having at least 75% sequence identity with it;

[0190] (17) The sense strand of the siRNA contains SEQ ID NO:38 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:80 or a nucleotide sequence having at least 75% sequence identity with it;

[0191] (18) The sense strand of the siRNA contains SEQ ID NO:39 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:81 or a nucleotide sequence having at least 75% sequence identity with it;

[0192] (19) The sense strand of the siRNA contains SEQ ID NO:40 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:82 or a nucleotide sequence having at least 75% sequence identity with it;

[0193] (20) The sense strand of the siRNA comprises SEQ ID NO:41 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA comprises SEQ ID NO:83 or a nucleotide sequence having at least 75% sequence identity with it; or

[0194] (21) The sense strand of the siRNA contains SEQ ID NO:42 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:84 or a nucleotide sequence having at least 75% sequence identity with it.

[0195] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:64 to 84 may, for example, contain a sequence identity of at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% with one of SEQ ID NO:1 to 21, respectively. Therefore, the antisense strand of the siRNA may contain a nucleotide sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with one of SEQ ID NO:64 to 84.

[0196] A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:64 to 84 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide substitutions (e.g., two or more, three or more, four or more, or five or more) relative to one of SEQ ID NO:64 to 84. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:64 to 84 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide deletions relative to one of SEQ ID NO:64 to 84. A nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:64 to 84 may respectively comprise: one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide insertions relative to one of SEQ ID NO:64 to 84.

[0197] Nucleic acid molecules may, for example, contain siRNA, which includes:

[0198] (1) such as Figure 1 The positive strand shown in C, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 1 The antisense strand shown in C or a nucleotide sequence having at least 75% sequence identity with it;

[0199] (2) For example Figure 1 The positive strand shown in D, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 1 The antisense strand shown in D or a nucleotide sequence having at least 75% sequence identity with it;

[0200] (3) such as Figure 1 E represents the positive strand or a nucleotide sequence having at least 75% sequence identity with it, and as shown in Figure E. Figure 1 E represents the antisense strand or a nucleotide sequence that has at least 75% sequence identity with it;

[0201] (4) such as Figure 1 The positive strand shown in F, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 1 The antisense strand shown in F or a nucleotide sequence that has at least 75% sequence identity with it;

[0202] (5) such as Figure 1 The positive strand shown in A, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 1 The antisense strand shown in A or a nucleotide sequence having at least 75% sequence identity with it;

[0203] (6) For example Figure 1 The positive strand shown in B, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 1 The antisense strand shown in B or a nucleotide sequence that has at least 75% sequence identity with it;

[0204] (7) For example Figure 2 The positive strand shown in A, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 2 The antisense strand shown in A or a nucleotide sequence having at least 75% sequence identity with it;

[0205] (8) such as Figure 2 The positive strand shown in B, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 2 The antisense strand shown in B or a nucleotide sequence that has at least 75% sequence identity with it;

[0206] (9) such as Figure 2 The positive strand shown in C, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 2 The antisense strand shown in C or a nucleotide sequence having at least 75% sequence identity with it;

[0207] (10) such as Figure 2 The positive strand shown in D, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 2 The antisense strand shown in D or a nucleotide sequence having at least 75% sequence identity with it;

[0208] (11) such as Figure 2 E represents the positive strand or a nucleotide sequence having at least 75% sequence identity with it, and as shown in Figure E. Figure 2 E represents the antisense strand or a nucleotide sequence that has at least 75% sequence identity with it;

[0209] (12) such as Figure 3 The positive strand shown in A, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 3 The antisense strand shown in A or a nucleotide sequence having at least 75% sequence identity with it;

[0210] (13) such as Figure 3 The positive strand shown in B, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 3 The antisense strand shown in B or a nucleotide sequence that has at least 75% sequence identity with it;

[0211] (14) such as Figure 3 The positive strand shown in C, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 3 The antisense strand shown in C or a nucleotide sequence having at least 75% sequence identity with it;

[0212] (15) such as Figure 3 The positive strand shown in D, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 3 The antisense strand shown in D or a nucleotide sequence having at least 75% sequence identity with it;

[0213] (16) such as Figure 3 E represents the positive strand or a nucleotide sequence having at least 75% sequence identity with it, and as shown in Figure E. Figure 3 E represents the antisense strand or a nucleotide sequence that has at least 75% sequence identity with it;

[0214] (17) such as Figure 4 The positive strand shown in A, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 4The antisense strand shown in A or a nucleotide sequence having at least 75% sequence identity with it;

[0215] (18) such as Figure 4 The positive strand shown in B, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 4 The antisense strand shown in B or a nucleotide sequence that has at least 75% sequence identity with it;

[0216] (19) such as Figure 4 The positive strand shown in C, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 4 The antisense strand shown in C or a nucleotide sequence having at least 75% sequence identity with it;

[0217] (20) such as Figure 4 The positive strand shown in D, or a nucleotide sequence having at least 75% sequence identity with it, and such as Figure 4 The antisense strand shown in D or a nucleotide sequence having at least 75% sequence identity with it; or

[0218] (21) As Figure 4 E represents the positive strand or a nucleotide sequence having at least 75% sequence identity with it, and as shown in Figure E. Figure 4 E represents the antisense strand or a nucleotide sequence that has at least 75% sequence identity with it.

[0219] and Figures 1 to 4 Any sequence shown has a nucleotide sequence with at least 75% sequence identity, for example, containing a sequence identity with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the sequence. Therefore, the sense and / or antisense strands of the siRNA may contain nucleotide sequences with at least 75% sequence identity. Figures 1 to 4 The sequences shown have nucleotide sequences with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity.

[0220] and Figures 1 to 4 The nucleotide sequence shown, having at least 75% sequence identity, may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide substitutions relative to that sequence. Figures 1 to 4 A nucleotide sequence having at least 75% sequence identity as the sequence shown may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide deletions relative to that sequence. Figures 1 to 4Nucleotide sequences with at least 75% sequence identity as shown may contain one or more (e.g., two or more, three or more, four or more, or five or more) nucleotide insertions relative to that sequence.

[0221] Generally, when siRNA enters a cell, it forms a complex called the RNA-Induced Silencing Complex (RISC). The RISC is a multi-protein complex containing a single strand of siRNA. The RISC uses the siRNA as a template to recognize complementary mRNA. When it finds the complementary strand, it activates RNase and cleaves the mRNA. siRNA typically contains overhangs at its 3' and / or 5' ends, and it is believed that such overhangs can assist in the interaction with the RISC and help maintain the siRNA and its function in vivo. Therefore, the nucleic acid silencing molecule of the present invention and / or the siRNA it contains may contain overhangs at its 3' and / or 5' ends.

[0222] For example, the 3' end of the sense strand of the siRNA may protrude one or more (e.g., two or more, three or more, four or more, or five or more) nucleotides beyond the 5' end of the antisense strand of the siRNA. For example, the 3' end of the sense strand of the siRNA may protrude two nucleotides beyond the 5' end of the antisense strand of the siRNA.

[0223] For example, the 3' end of the antisense strand of the siRNA may protrude one or more (e.g., two or more, three or more, four or more, or five or more) nucleotides beyond the 5' end of the sense strand of the siRNA. For example, the 3' end of the antisense strand of the siRNA may protrude two nucleotides beyond the 5' end of the sense strand of the siRNA.

[0224] For example, the 3' end of the sense strand of the siRNA may protrude one or more (e.g., two or more, three or more, four or more, or five or more) nucleotides beyond the 5' end of the antisense strand of the siRNA, and the 3' end of the antisense strand of the siRNA may, for example, protrude one or more (e.g., two or more, three or more, four or more, or five or more) nucleotides beyond the 5' end of the sense strand of the siRNA. For example, the 3' end of the sense strand of the siRNA may protrude two nucleotides beyond the 5' end of the antisense strand of the siRNA, and the 3' end of the antisense strand of the siRNA may protrude two nucleotides beyond the 5' end of the sense strand of the siRNA.

[0225] However, in some cases, it is advantageous for siRNA to lack a protrusion at the 3' end of the sense strand. Therefore, in some preferred aspects of this disclosure, the 3' end of the sense strand of the siRNA does not protrude beyond the 5' end of the antisense strand.

[0226] Exemplary protrusions are shown Figures 1 to 4 middle.

[0227] Adhesion

[0228] Nucleic acid silencing molecules can be conjugated to one or more non-nucleic acid moieties. For example, a nucleic acid silencing molecule can be conjugated to two or more, three or more, four or more, or five or more non-nucleic acid moieties.

[0229] Preferably, the non-nucleic acid portion is the delivery portion. A delivery portion is a molecule that assists in the delivery of nucleic acid silencing molecules in vivo. For example, a delivery portion can facilitate the delivery of nucleic acid silencing molecules to target organs, tissues, or cell types. Any non-nucleic acid portion mentioned below can serve as a delivery portion.

[0230] The non-nucleic acid portion may, for example, be hydrophobic. The non-nucleic acid portion may, for example, contain lipids. For instance, the non-nucleic acid portion may contain cholesterol, fatty acids, triglycerides, or phospholipids. Fatty acids may be saturated or unsaturated. Unsaturated fatty acids may be polyunsaturated. Preferably, the non-nucleic acid portion contains cholesterol or polyunsaturated fatty acids.

[0231] The non-nucleic acid portion may, for example, contain peptides, polypeptides, or proteins. For instance, the non-nucleic acid portion may contain peptide ligands. Peptides or peptide ligands may, for example, be homologous ligands of receptors present on the surface of target cells or contained on the cell surface of target tissues or organs. The non-nucleic acid portion may, for example, contain antibodies or antibody fragments. Antibodies or antibody fragments may, for example, be capable of binding to antigens present on the surface of target cells or contained on the cell surface of target tissues or organs. Antibody fragments may, for example, contain scFv, Fab, modified Fab, Fab', modified Fab', F(ab')2, or scFv2, or be composed of scFv, Fab, modified Fab, Fab', modified Fab', F(ab')2, or scFv2.

[0232] The non-nucleic acid portion may, for example, contain sugars, disaccharides, or polysaccharides. The non-nucleic acid portion may, for example, contain amino sugars. Preferably, the non-nucleic acid portion contains N-acetylgalactosamine (GalNAc). GalNAc is capable of binding to the asialic acid glycoprotein receptor (ASGPR) expressed on the surface of hepatocytes. Therefore, by conjugating nucleic acid silencing molecules to GalNAc, it is possible to specifically deliver nucleic acid silencing molecules to hepatocytes. GalNAc is highly potent and has been shown to significantly increase the uptake of nucleic acid silencing molecules by hepatocytes and prolong their duration. Studies in mice have also shown that GalNAc-conjugated oligonucleotides have some biodistribution in the kidneys.

[0233] The non-nucleic acid portion may, for example, comprise nanoparticles. Suitable nanoparticles are known in the art. Methods for producing such nanoparticles are also known. Nanoparticles may be, for example, lipid nanoparticles, liposomes, polymer nanoparticles, inorganic nanoparticles, virus-like particles (VLPs), self-assembled proteins, calcium phosphate nanoparticles, silicon nanoparticles, or gold nanoparticles. Preferably, the nanoparticles are lipid nanoparticles. Conjugating nucleic acid silencing molecules to lipid nanoparticles can enhance the biodistribution and uptake of nucleic acid silencing molecules in the liver and kidneys.

[0234] Treatment methods and medical uses

[0235] This article discloses a method for treating a disease in a subject, comprising administering to the subject a composition comprising a nucleic acid silencing molecule disclosed herein.

[0236] This document also discloses a composition for use in a method of treating a disease in a subject, wherein the composition comprises a nucleic acid silencing molecule of the present disclosure, and the method comprises administering the nucleic acid silencing molecule to the subject.

[0237] Nucleic acid silencing molecules

[0238] The treatment methods and medical uses described herein include administering to a subject a composition comprising a nucleic acid silencing molecule that reduces STAT6 expression. Such nucleic acid silencing molecules are described in detail above. Any aspect of the above description in conjunction with the nucleic acid silencing molecules of this disclosure is equally applicable to the treatment methods or medical uses of this disclosure.

[0239] The composition may contain one or more nucleic acid silencing molecules that reduce STAT6 expression. For example, the composition may contain two or more, five or more, ten or more, 20 or more, 50 or more, 100 or more, 200 or more, 500 or more, 1000 or more, 2000 or more, 5000 or more, 10000 or more, 20000 or more, 50000 or more, 100000 or more, 200000 or more, 500000 or more, 1000000 or more, 2000000 or more, 5000000 or more, 1 x 10 7 or more, 2 x 10 7 or more, 5 x 10 7 or more, 1 x 10 8 or more, 2 x 10 8 or more, 5 x 10 8 or more, 1 x 10 9 or more, 2 x 10 9 More or more, or 5 x 10 9 One or more nucleic acid silencing molecules that reduce STAT6 expression. Preferably, all the multiple nucleic acid silencing molecules contained in a single-dose composition contain the same nucleotide sequence.

[0240] In one aspect, the nucleic acid silencing molecule may comprise one or more nucleotide phosphate thioesters, or be composed of nucleotide phosphate thioesters. When the composition comprises two or more such nucleic acid silencing molecules, each of the two or more nucleic acid silencing molecules may be the same stereoisomer. Preferably, the composition does not contain other nucleic acid silencing molecules. This ensures the stereopurity of the composition. The stereopurity of nucleic acid silencing molecules has been described in the art, for example in Iwamoto et al. (2017), Nature Biotechnology, 35:9, 845-851.

[0241] Reduce STAT6 expression

[0242] The nucleic acid silencing molecule contained in the composition reduces STAT6 expression. The reduction in STAT6 expression has been described in detail above in conjunction with the nucleic acid silencing molecule of this disclosure. Any aspect described in conjunction with the nucleic acid silencing molecule of this disclosure is equally applicable to the therapeutic methods or medical uses of this disclosure.

[0243] disease

[0244] Reducing STAT6 expression can treat many conditions. For example, as mentioned above, STAT6 is highly expressed in various cancers, where it regulates the expression of genes involved in immune responses, cell survival, tumor proliferation, and metastasis. Therefore, reducing STAT6 expression in cancer cells can kill cancer cells, reduce or prevent tumor growth, and reduce or prevent metastasis. Furthermore, it has been demonstrated that STAT6 expression by immune cells in the tumor microenvironment contributes to the formation of an immunosuppressive microenvironment conducive to tumor maintenance and growth. Reducing STAT6 expression in the tumor microenvironment can promote the formation of a pro-inflammatory microenvironment conducive to killing tumor cells.

[0245] Lowering STAT6 expression may also be beneficial in diseases caused by or associated with an overactive Th2 response. For example, Th2 responses are associated with inflammatory diseases, allergic diseases, autoimmune diseases, and graft-versus-host disease (GVHD). Since STAT6 is a key signaling transduction molecule for inducing and maintaining Th2 responses, lowering STAT6 expression in subjects can reduce the quantity and / or activity of Th2 responses. Therefore, it can reduce or eliminate clinical signs or symptoms of an overactive Th2 response and may halt the progression of clinical signs or symptoms.

[0246] Therefore, the disease to be treated can be cancer. The cancer can be, for example, a cancer in which cells express or overexpress STAT6. The cancer can be, for example, a solid tumor. For example, the cancers mentioned can be anal cancer, cholangiocarcinoma, bladder cancer, leukemia, bone cancer, bowel cancer, brain tumors, breast cancer, colorectal cancer, cervical cancer, endocrine tumors, eye cancer (e.g., ocular melanoma), fallopian tube cancer, gallbladder cancer, head and neck cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, liver cancer, lung cancer, lymph node cancer, melanoma, mesothelioma, myeloma, neuroendocrine tumors, ovarian cancer, esophageal cancer, pancreatic cancer, penile cancer, primary peritoneal cancer, prostate cancer, Pseudomyxoma peritonei, skin cancer, small intestine cancer, soft tissue sarcoma, spinal cord tumors, stomach cancer, testicular cancer, thymic cancer, thyroid cancer, tracheal cancer, cancer of unknown primary origin, vaginal cancer, vulvar cancer, or endometrial cancer. The cancers mentioned can also be, for example, leukemia, myeloma, or lymphoma. The cancer can be primary or secondary.

[0247] Alternatively, the disease can be one in which the subject may benefit from a reduced or relatively reduced antigen-specific Th2 response. A relatively reduced antigen-specific Th2 response can, for example, refer to a shift in the balance of antigen-specific T cell responses from Th2. For example, the balance may shift to one or more other T helper cell subsets (e.g., Th1, Th17, Treg, and Tfh).

[0248] The disease mentioned can be, for example, an inflammatory disease. A prime example of an inflammatory disease is asthma. Therefore, the disease could be asthma. Other inflammatory diseases include ankylosing spondylitis, antiphospholipid syndrome, chronic relapsing multifocal osteomyelitis, gout, Henoch-Schonlein purpura, dermatomyositis, idiopathic arthritis, scleroderma, Kawasaki disease, mixed connective tissue disease, myositis, post-streptococcal inflammatory syndrome, psoriatic arthritis, reactive arthritis, scleroderma, spondyloarthritis, systemic juvenile idiopathic arthritis, undifferentiated connective tissue disease, uveitis, and vasculitis.

[0249] The disease may be, for example, an allergic disease, such as atopic dermatitis, allergic airway inflammation, or allergic rhinitis.

[0250] The diseases mentioned may include, for example, autoimmune conditions such as alopecia areata, autoimmune encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, dermatomyositis, type 1 diabetes mellitus, autoimmune juvenile idiopathic arthritis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, idiopathic thrombocytopenic purpura, myasthenia gravis, autoimmune myocarditis, multiple sclerosis, pemphigus / pemphigoid, pernicious anemia, polyarteritis nodosa, polymyositis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, scleroderma / systemic sclerosis, Sjögren's syndrome, systemic lupus erythematosus, autoimmune thyroiditis, uveitis, or vitiligo.

[0251] The disease could be, for example, GVHD.

[0252] Subjects

[0253] The subject may be a mammal, for example. The mammal may be a human or a non-human mammal, such as a dog, cat, horse, or farm animal. Preferably, the subject is a human.

[0254] Subjects may be, for example, adults. Subjects may be, for example, minors.

[0255] Application and formulation

[0256] The composition can be administered via any route. Suitable routes include, but are not limited to, intravenous, intratracheal, intranasal, intrathecal, intravenous, intramuscular, intraperitoneal, subcutaneous, intradermal, transdermal, and oral / buccal routes.

[0257] The composition may be contained in a delivery vector optimized for in vivo delivery of nucleic acid silencing molecules. Suitable delivery vectors are known in the art, including, for example, cell-targeting portions, cell-penetrating portions, lipids, lipoproteins, liposomes, lipid complexes, peptides, GalNAc, antibodies, aptamers, nanoparticles, exosomes, globular nucleic acids, and DNA cages.

[0258] The composition can be prepared with a physiologically acceptable carrier or diluent. Typically, such compositions are prepared as liquid suspensions of nucleic acid silencing molecules and / or nucleic acid silencing molecules linked to a delivery carrier. The nucleic acid silencing molecules and / or nucleic acid silencing molecules linked to a delivery carrier can be mixed with pharmaceutically acceptable excipients compatible with the active ingredient. Suitable excipients include, for example, water, saline, dextran, glycerol, analogs, and combinations thereof. Furthermore, if desired, the pharmaceutical composition may contain small amounts of excipients, such as wetting agents or emulsifiers, and / or pH buffers.

[0259] Nucleic acid silencing molecules and / or nucleic acid silencing molecules linked to a delivery vector are administered in a manner compatible with the dosage formulation, and the amount will be therapeutically effective. The amount to be administered depends on the subject to be treated, the disease to be treated, and the capacity of the subject's immune system. The precise amount of nucleic acid silencing molecules and / or nucleic acid silencing molecules linked to a delivery vector to be administered may depend on the judgment of the physician and may vary from subject to subject.

[0260] Combination therapy

[0261] The composition may be administered as part of a combination therapy. That is, the treatment method or medical use may include administering additional therapeutic compositions or treatment regimens to the subject. For example, when nucleic acid silencing molecules reduce rather than eliminate STAT6 expression, additional compositions or treatment regimens may be required. However, in some cases, reducing (rather than eliminating) STAT6 expression may be sufficient to treat the disease.

[0262] The composition can be administered as part of a combination therapy, in combination with any available therapeutic composition or regimen targeting a specific disease.

[0263] For example, when the disease is cancer, the composition can be used in combination with (i) surgical resection, (ii) radiation therapy and / or (iii) systemic therapy. For example, the composition can be used in combination with: (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii).

[0264] Systemic therapy may include or consist of (a) chemotherapy. Systemic therapy may include or consist of (b) immunotherapy. Systemic therapy may include or consist of (c) targeted therapy. For example, systemic therapy may include or consist of: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c).

[0265] Chemotherapy is well known in the art. Such chemotherapy may, for example, contain platinum-based antitumor drugs, such as cisplatin or carboplatin. Such chemotherapy may, for example, contain antimetabolites, such as fluorouracil (5-FU), gemcitabine, or methotrexate. Such chemotherapy may, for example, contain taxanes, such as docetaxel or paclitaxel. Such chemotherapy may, for example, contain anthracyclines, such as doxorubicin. Such chemotherapy may, for example, contain vinblastine alkaloids, such as vinca alkaloids. Such chemotherapy may, for example, contain antitumor antibiotics, such as mitomycin. Such chemotherapy may, for example, contain alkylating agents, such as ifosfamide.

[0266] Immunotherapy is well known in the art. Such immunotherapies include, for example, therapeutic immune cells, immunomodulators, checkpoint inhibitors, and vaccines. Therapeutic immune cells may include T cells, such as engineered T cells like CAR T cells or T cells expressing engineered TCRs. Immunomodulators may include, for example, interleukins, cytokines, chemokines, and immunomodulatory imide drugs. Immunomodulators may include, for example, Multikine (an interleukin injection). Immunomodulators may include, for example, monalizumab, a humanized anti-NKG2A blocking antibody that prevents the suppression of CD8+ T cells and NK cells by HLA-E-expressing tumor cells. Checkpoint inhibitors may include, for example, CTLA-4 inhibitors or PD-1 axis binding antagonists. PD-1 axis binding antagonists may include, for example, pembrolizumab, nivolumab, avelumab, and atezolizumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in detail below. Vaccines may include, for example, vidutolimod (CMP-001), a Toll-like receptor 9 (TLR9) agonist cancer vaccine.

[0267] Targeted therapy is well known in the art. The term targeted therapy is a term used in this art to refer to treatments that target specific genes and proteins that help cancer cells survive and grow. Targeted therapies may include, for example, fibroblast growth factor receptor inhibitors (such as erdatinib) and antibody-drug conjugates (such as enfortumab vedotin-ejfv or sacituzumab govitecan). Enfortumab is a conjugate of an antibody targeting nectin-4 and a microtubule inhibitor. Sacituzumab is a conjugate of an antibody targeting Trop-2 and a topoisomerase inhibitor.

[0268] When the disease is one in which the subject may benefit from a reduced or relatively reduced antigen-specific Th2 response, the composition may be used, for example, in combination with an immunosuppressive drug. Other drugs that may be useful in combination therapy may depend on the disease to be treated. Treatment for inflammatory diseases, allergic diseases, autoimmune diseases, and GVHD is well known in the art and can be used in combination with the nucleic acid silencing molecules of this disclosure.

[0269] In combination therapy, the composition containing a nucleic acid silencing molecule that reduces STAT6 expression is administered in an amount that would be therapeutically effective when administered in combination with another therapeutic composition or regimen. The other therapeutic composition is administered in an amount that would be therapeutically effective when administered in combination with the composition containing a nucleic acid silencing molecule that reduces STAT6 expression. The composition containing a nucleic acid silencing molecule that reduces STAT6 expression and the other therapeutic composition may be administered together, for example, simultaneously. The composition containing a nucleic acid silencing molecule that reduces STAT6 expression and the other therapeutic composition may be administered separately, for example, at different times. For example, the composition containing a nucleic acid silencing molecule that reduces STAT6 expression may be administered before the other therapeutic composition. The composition containing a nucleic acid silencing molecule that reduces STAT6 expression may be administered after the other therapeutic composition. The administration of the composition containing a nucleic acid silencing molecule that reduces STAT6 expression may be alternated with the administration of the other therapeutic composition.

[0270] In another aspect of the combination therapy, the composition comprising a nucleic acid silencing molecule that reduces STAT6 expression is administered in an amount that would be therapeutically effective when combined with an additional treatment regimen. The treatment regimen is implemented to such an extent that it would be therapeutically effective when combined with the composition comprising a nucleic acid silencing molecule that reduces STAT6 expression. The composition comprising a nucleic acid silencing molecule that reduces STAT6 expression may be administered before, during, or after the implementation of the treatment regimen. Preferably, the composition comprising a nucleic acid silencing molecule that reduces STAT6 expression is administered during the implementation of the treatment regimen.

[0271] In vitro methods

[0272] As described above, the nucleic acid silencing molecules disclosed herein can be used to reduce STAT6 expression in vitro. For example, nucleic acid silencing molecules can be used as research tools, such as for studying cancer, inflammatory diseases, allergic diseases, or autoimmune diseases. Therefore, this disclosure provides an in vitro method for reducing STAT6 expression in cells, comprising contacting cells with the nucleic acid silencing molecules of this disclosure.

[0273] The cell can be any type of cell. For example, the cell can originate from any tissue. The cell can originate from any species. Preferably, the cell is human. The cell can be a healthy cell or a diseased cell. A diseased cell can be, for example, a cancer cell. The cell can be a naturally occurring cell. The cell can originate from a cell line.

[0274] The mechanisms for contacting cells with nucleic acid silencing molecules are well known in the art. For example, contact can occur in the wells of a microplate or in other types of containers such as cell culture flasks or test tubes. The contact step can be performed before or simultaneously with cell culture. The conditions required for culturing various cell types are well known in the art.

[0275] In addition to nucleic acid silencing molecules, cells may also come into contact with one or more other molecules. This additional molecule may be another nucleic acid silencing molecule targeting STAT6. This additional molecule may be a nucleic acid silencing molecule targeting genes other than STAT6, such as different genes involved in Th2 induction and maintenance, or another gene overexpressed in cancer cells. Preferably, this additional molecule can promote the reduction or elimination of STAT6 expression by the nucleic acid silencing molecule.

[0276] Example

[0277] The following examples illustrate this disclosure.

[0278] Example 1

[0279] To address the need for therapies that effectively reduce STAT6 expression, siRNAs targeting STAT6 have been developed. These siRNAs show... Figures 1 to 4 The sequence of the sense strand of each siRNA is also shown in Table 1. The sequence of the antisense strand of each siRNA is shown in Table 2. In each case, “base sequence” refers to the unmodified nucleotide sequence of the strand. “Modified sequence” shows the specific combination and position of modified (2'-OMe, 2'-F, thiophosphorylation) nucleotides used for optimal repression of STAT6 expression.

[0280] Table 1: The positive strand of the siRNA disclosed in this paper

[0281]

[0282]

[0283] d T = T DNA

[0284] N N = thiophosphate

[0285] m C m A, m G, m U = 2'-OMe modified nucleotides

[0286] f A, f C f G, f U = 2'-F modified nucleotides

[0287] Table 2: Antisense strand of the siRNA disclosed herein

[0288]

[0289]

[0290] d T = T DNA

[0291] N N = thiophosphate

[0292] m C m A, m G, m U = 2'-OMe modified nucleotides

[0293] f A, f C f G, f U = 2'-F modified nucleotides

[0294] At least the siRNAs in family 1 (i.e., siRNA 1, 1w, 1a, 1b, 1c, and 1d) possess advantageous properties. In particular, siRNAs in family 1 have been shown to slow tumor growth. Therefore, they could be used to treat cancers, such as those expressing STAT6.

[0295] Furthermore, siRNAs in family 1 target human and mouse STAT6. Therefore, they can be used in xenograft tumor models to block STAT6, whether it is produced by human cancer cells or by mouse stromal cells in the tumor microenvironment. They can also be used in syngeneic tumor models in which all STAT6 is entirely mouse-derived; or in mouse or rat inflammation models involving only rodent STAT6.

[0296] Example 2

[0297] The potency of the siRNA designed in Example 1 was tested in vitro to evaluate its ability to reduce STAT6 expression. For example, the potency was studied by measuring the expression level of STAT6 mRNA by RT-qPCR over a biologically relevant period following transfection of cultured cells. Further, for example, the potency was studied by measuring the expression level of STAT6 protein in transfected cells using Western blot analysis over a biologically relevant period following transfection.

[0298] In more detail, the potency of siRNAs 1, 1w, 1a, 1b, 1c, 1d, 2, 2a, 2b, 2c, 2d, 3, 3a, 3b, 3c, 3d, 4, 4a, 4b, 4c, and 4d (in both base sequence and modified forms) was evaluated in A549 lung adenocarcinoma cells, THP-1 cells, and peripheral blood mononuclear cells (PBMCs). A549 cells are alveolar epithelial cells that respond to IL4 and IL13 to activate STAT6. They mimic the inflammatory response in the lungs during asthma. They also mimic cancer. THP-1 is a mononuclear cell line that mimics inflammatory diseases.

[0299] Each cell type was transfected with one of the siRNAs in (a) the absence of lipofectamine, and in a separate reaction, in (b) the presence of lipofectamine. Transfection was performed in at least triplicate.

[0300] Cells were stimulated with varying concentrations of IL-4 and / or IL-13 over a period of time. Specifically, cells were stimulated with IL-4. STAT6 expression was then measured at an endpoint. Specifically, STAT6 expression was measured at the messenger level, and STAT-6 mRNA was quantified by RT-PCR. Total STAT6 expression was also measured at the protein level by Western blot analysis. Activated STAT6 was measured by Western blot analysis using a phosphorylated STAT6 Tyr641 antibody to detect phosphorylated STAT6 protein. STAT6 activity was indirectly measured by assessing the expression of the cell surface protein CD23 (e.g., by flow cytometry or immunofluorescence imaging) or by assessing CCL17 secretion via immunoassay.

[0301] They also performed functional assays of stimulated cells to quantify cell proliferation and / or apoptosis.

[0302] The tested siRNAs, particularly those containing 2'-OMe and / or 2'-F modifications, effectively reduced STAT6 expression levels. Transfection with siRNAs, especially those containing 2'-OMe and / or 2'-F modifications, resulted in decreased proliferation and / or increased apoptosis. Therefore, the tested siRNAs demonstrated surprisingly better efficacy than known and / or unmodified siRNA sequences, particularly in reducing STAT6 expression levels, decreasing proliferation, and / or increasing apoptosis.

[0303] Example 3

[0304] Further testing (in both base sequence and modified forms) of siRNAs 1, 1w, 1a, 1b, 1c, 1d, 2, 2a, 2b, 2c, 2d, 3, 3a, 3b, 3c, 3d, 4, 4a, 4b, 4c, and 4d was conducted in HT-29 cells. HT-29 is a human colorectal adenocarcinoma cell line with an epithelial morphology. STAT6 is strongly expressed in various tumors, with the highest expression levels in malignant lymphomas, as well as pancreatic cancer, colorectal cancer, prostate cancer, and breast cancer. STAT6 expression in colorectal cancer is associated with increased malignancy, poor prognosis, and decreased survival.

[0305] HT-29 cells were transfected with one of the siRNAs in (a) the absence of liposomes, and in separate transfections, in (b) the presence of liposomes. Transfections were performed in at least triplicate.

[0306] STAT6 expression was measured after a biologically relevant period. Specifically, STAT-6 mRNA was quantified by RT-PCR to measure STAT6 expression at the messenger level. STAT6 expression was also measured at the protein level by Western blotting. Western blotting can measure total STAT6, or phosphorylated STAT6 can be measured using a phosphorylated-STAT6 Tyr641 antibody as the detection antibody.

[0307] Functional assays were also performed on transfected cells, measuring proliferation and / or apoptosis at different time points.

[0308] The tested siRNAs, particularly those containing 2'-OMe and / or 2'-F modifications, effectively reduced STAT6 expression levels. Transfection with siRNAs, especially those containing 2'-OMe and / or 2'-F modifications, resulted in decreased proliferation and / or increased apoptosis. Therefore, the tested siRNAs demonstrated surprisingly better efficacy than known and / or unmodified siRNA sequences, particularly in reducing STAT6 expression levels, decreasing proliferation, and / or increasing apoptosis.

Claims

1. A nucleic acid silencing molecule comprising or composed of small interfering RNA (siRNA) that reduces the expression of signal transduction and activator of transcription 6 (STAT6), wherein, The siRNA targets exons 5, 15, 19, or 10 of STAT6.

2. The nucleic acid silencing molecule according to claim 1, wherein, The positive strand of the siRNA comprises any one of SEQ ID NO:5, 3, 4, 6, 8 to 11, 13 to 16 and 18 to 21 or a nucleotide sequence having at least 75% sequence identity with any one of SEQ ID NO:5, 3, 4, 6, 8 to 11, 13 to 16 and 18 to 21, optionally wherein the positive strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90% or at least 95% sequence identity with any one of SEQ ID NO:5, 3, 4, 6, 8 to 11, 13 to 16 and 18 to 21.

3. The nucleic acid silencing molecule according to claim 1 or 2, wherein: (a) The sense strand of the siRNA contains SEQ ID NO:5 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:47 or a nucleotide sequence having at least 75% sequence identity with it; (b) The sense strand of the siRNA contains SEQ ID NO:3 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:45 or a nucleotide sequence having at least 75% sequence identity with it; (c) The sense strand of the siRNA contains SEQ ID NO:4 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:46 or a nucleotide sequence having at least 75% sequence identity with it; (d) The sense strand of the siRNA contains SEQ ID NO:6 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:48 or a nucleotide sequence having at least 75% sequence identity with it; (e) The sense strand of the siRNA contains SEQ ID NO:8 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:50 or a nucleotide sequence having at least 75% sequence identity with it; (f) The sense strand of the siRNA contains SEQ ID NO:9 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:51 or a nucleotide sequence having at least 75% sequence identity with it; (g) The sense strand of the siRNA contains SEQ ID NO:10 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:52 or a nucleotide sequence having at least 75% sequence identity with it; (h) The sense strand of the siRNA contains SEQ ID NO:11 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:53 or a nucleotide sequence having at least 75% sequence identity with it; (i) The sense strand of the siRNA contains SEQ ID NO:13 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:55 or a nucleotide sequence having at least 75% sequence identity with it; (j) The sense strand of the siRNA contains SEQ ID NO:14 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:56 or a nucleotide sequence having at least 75% sequence identity with it; (k) The sense strand of the siRNA contains SEQ ID NO:15 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:57 or a nucleotide sequence having at least 75% sequence identity with it; (l) The sense strand of the siRNA contains SEQ ID NO:16 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:58 or a nucleotide sequence having at least 75% sequence identity with it; (m) The sense strand of the siRNA contains SEQ ID NO:18 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:60 or a nucleotide sequence having at least 75% sequence identity with it; (n) The sense strand of the siRNA contains SEQ ID NO:19 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:61 or a nucleotide sequence having at least 75% sequence identity with it; (o) The sense strand of the siRNA comprises SEQ ID NO:20 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA comprises SEQ ID NO:62 or a nucleotide sequence having at least 75% sequence identity with it; or (p) The sense strand of the siRNA contains SEQ ID NO:21 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:63 or a nucleotide sequence having at least 75% sequence identity with it.

4. The nucleic acid silencing molecule according to claim 3, wherein, The antisense strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90%, or at least 95% sequence identity with one of the corresponding sequences in SEQ ID NO:45 to 48, 50 to 53, 55 to 58, and 60 to 63.

5. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The nucleic acid silencing molecule comprises one or more nucleotides modified with 2'-O-methyl (2'-OMe) or consists of nucleotides modified with 2'-O-methyl (2'-OMe).

6. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The nucleic acid silencing molecule comprises one or more 2'-fluorine (2'-F) modified nucleotides or is composed of 2'-fluorine (2'-F) modified nucleotides.

7. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The nucleic acid silencing molecule comprises or is composed of one or more nucleoside thiophosphates.

8. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The nucleic acid silencing molecule contains a nucleotide sequence of about 15 to about 30 nucleotides in length, optionally about 20 to about 25 nucleotides in length.

9. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The positive strand of the siRNA comprises any one of SEQ ID NO: 24 to 27, 29 to 32, 34 to 37 and 39 to 42, or a nucleotide sequence having at least 75% sequence identity with it.

10. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The positive strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO:24 to 27, 29 to 32, 34 to 37, and 39 to 42.

11. The nucleic acid silencing molecule according to any one of the preceding claims, wherein: (a) The sense strand of the siRNA contains SEQ ID NO:26 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:68 or a nucleotide sequence having at least 75% sequence identity with it; (b) The sense strand of the siRNA contains SEQ ID NO:24 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:66 or a nucleotide sequence having at least 75% sequence identity with it; (c) The sense strand of the siRNA contains SEQ ID NO:25 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:67 or a nucleotide sequence having at least 75% sequence identity with it; (d) The sense strand of the siRNA contains SEQ ID NO:27 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:69 or a nucleotide sequence having at least 75% sequence identity with it; (e) The sense strand of the siRNA contains SEQ ID NO:29 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:71 or a nucleotide sequence having at least 75% sequence identity with it; (f) The sense strand of the siRNA contains SEQ ID NO:30 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:72 or a nucleotide sequence having at least 75% sequence identity with it; (g) The sense strand of the siRNA contains SEQ ID NO:31 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:73 or a nucleotide sequence having at least 75% sequence identity with it; (h) The sense strand of the siRNA contains SEQ ID NO:32 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:74 or a nucleotide sequence having at least 75% sequence identity with it; (i) The sense strand of the siRNA contains SEQ ID NO:34 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:76 or a nucleotide sequence having at least 75% sequence identity with it; (j) The sense strand of the siRNA contains SEQ ID NO:35 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:77 or a nucleotide sequence having at least 75% sequence identity with it; (k) The sense strand of the siRNA contains SEQ ID NO:36 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:78 or a nucleotide sequence having at least 75% sequence identity with it; (l) The sense strand of the siRNA contains SEQ ID NO:37 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:79 or a nucleotide sequence having at least 75% sequence identity with it; (m) The sense strand of the siRNA contains SEQ ID NO:39 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:81 or a nucleotide sequence having at least 75% sequence identity with it; (n) The sense strand of the siRNA contains SEQ ID NO:40 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:82 or a nucleotide sequence having at least 75% sequence identity with it; (o) The sense strand of the siRNA contains SEQ ID NO:41 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:83 or a nucleotide sequence having at least 75% sequence identity with it; or (p) The sense strand of the siRNA contains SEQ ID NO:42 or a nucleotide sequence having at least 75% sequence identity with it, and the antisense strand of the siRNA contains SEQ ID NO:84 or a nucleotide sequence having at least 75% sequence identity with it.

12. The nucleic acid silencing molecule according to claim 11, wherein, The antisense strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90%, or at least 95% sequence identity with one of the corresponding sequences in SEQ ID NO: 66 to 69, 71 to 74, 76 to 79, and 81 to 84.

13. The nucleic acid silencing molecule according to any one of the preceding claims, wherein, The nucleic acid silencing molecule is conjugated to one or more non-nucleic acid moieties.

14. The nucleic acid silencing molecule according to any one of claims 3(c), 3(a), 3(d), 3(f), 3(g), 3(h), 3(j), 3(k), 3(l), 3(n), 3(o), 3(p), 11(c), 11(a), 11(d), 11(f), 11(g), 11(h), 11(j), 11(k), 11(l), 11(n), 11(o), and 11(p), wherein, The 3' end of the sense strand of the siRNA does not protrude beyond the 5' end of the antisense strand of the siRNA.

15. An in vitro method for reducing STAT6 expression in cells, comprising contacting the cells with a nucleic acid silencing molecule according to any one of the preceding claims.

16. A method of treating a disease in a subject, comprising administering to the subject a composition comprising a nucleic acid silencing molecule according to any one of claims 1 to 14.

17. The method according to claim 16, wherein, The disease in question is cancer.

18. The method according to claim 16, wherein, The disease is one in which the subject can benefit from a reduced or relatively reduced antigen-specific Th2 response.

19. The method according to claim 16 or 18, wherein, The disease in question is an inflammatory disease.

20. The method according to claim 16 or 18, wherein, The disease in question is an allergic disease.

21. The method according to claim 16 or 18, wherein, The disease in question is an autoimmune disease.

22. The method according to claim 16 or 18, wherein, The disease in question is graft-versus-host disease (GVHD).

23. A composition comprising a nucleic acid silencing molecule according to any one of claims 1 to 14, for use in the method according to any one of claims 16 to 22.

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