Search

CN-122003500-A - STAT6 siRNA

CN122003500ACN 122003500 ACN122003500 ACN 122003500ACN-122003500-A

Abstract

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

Inventors

  • G. ROBERSON
  • S Sultan

Assignees

  • 罗克福治疗有限公司

Dates

Publication Date
20260508
Application Date
20240805
Priority Date
20230804

Claims (20)

  1. 1. A nucleic acid silencing molecule comprising or consisting of a small interfering RNA (siRNA) that reduces the expression of signal transduction and transcription activator 6 (STAT 6), wherein the siRNA targets exons 5, 15, 19 or 10 of STAT 6.
  2. 2. The nucleic acid silencing molecule of claim 1, wherein the sense strand of the siRNA comprises a nucleotide sequence of any one of SEQ ID NOs 5, 3, 4, 6, 8 to 11, 13 to 16 and 18 to 21 or having at least 75% sequence identity thereto, optionally wherein the sense strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90% or at least 95% sequence identity to any one of SEQ ID NOs 5, 3, 4, 6, 8 to 11, 13 to 16 and 18 to 21.
  3. 3. The nucleic acid silencing molecule of claim 1 or 2, wherein: (a) The sense strand of the siRNA comprises SEQ ID NO 5 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 47 or a nucleotide sequence having at least 75% sequence identity thereto; (b) The sense strand of the siRNA comprises SEQ ID NO 3 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 45 or a nucleotide sequence having at least 75% sequence identity thereto; (c) The sense strand of the siRNA comprises SEQ ID NO. 4 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 46 or a nucleotide sequence having at least 75% sequence identity thereto; (d) The sense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 6 or having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 48 or having at least 75% sequence identity thereto; (e) The sense strand of the siRNA comprises a nucleotide sequence of SEQ ID No.8 or having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 50 or having at least 75% sequence identity thereto; (f) The sense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 9 or having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 51 or having at least 75% sequence identity thereto; (g) The sense strand of the siRNA comprises SEQ ID NO 10 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 52 or a nucleotide sequence having at least 75% sequence identity thereto; (h) The sense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 11 or having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises a nucleotide sequence of SEQ ID No. 53 or having at least 75% sequence identity thereto; (i) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 13 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 55 or at least 75% sequence identity thereto; (j) The sense strand of the siRNA comprises SEQ ID NO 14 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 56 or a nucleotide sequence having at least 75% sequence identity thereto; (k) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 15 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 57 or at least 75% sequence identity thereto; (l) The sense strand of the siRNA comprises SEQ ID NO. 16 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 58 or a nucleotide sequence having at least 75% sequence identity thereto; (m) the sense strand of said siRNA comprises the nucleotide sequence of SEQ ID No. 18 or at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises the nucleotide sequence of SEQ ID No. 60 or at least 75% sequence identity thereto; (n) the sense strand of said siRNA comprises SEQ ID No. 19 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises SEQ ID No. 61 or a nucleotide sequence having at least 75% sequence identity thereto; (o) the sense strand of the siRNA comprises SEQ ID NO. 20 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 62 or a nucleotide sequence having at least 75% sequence identity thereto, or (P) the sense strand of said siRNA comprises SEQ ID NO. 21 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises SEQ ID NO. 63 or a nucleotide sequence having at least 75% sequence identity thereto.
  4. 4. The nucleic acid silencing molecule of 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 to a corresponding one of SEQ ID NOs 45 to 48, 50 to 53, 55 to 58 and 60 to 63.
  5. 5. The nucleic acid silencing molecule of any of the preceding claims, wherein the nucleic acid silencing molecule comprises or consists of one or more 2 '-O-methyl (2' -OMe) modified nucleotides.
  6. 6. The nucleic acid silencing molecule of any of the preceding claims, wherein the nucleic acid silencing molecule comprises or consists of one or more 2 '-fluoro (2' -F) modified nucleotides.
  7. 7. The nucleic acid silencing molecule of any of the preceding claims, wherein the nucleic acid silencing molecule comprises or consists of one or more nucleoside phosphorothioates.
  8. 8. The nucleic acid silencing molecule of any of the preceding claims, wherein the nucleotide sequence comprised in the nucleic acid silencing molecule is about 15 to about 30 nucleotides in length, optionally about 20 to about 25 nucleotides in length.
  9. 9. The nucleic acid silencing molecule of any of the preceding claims, wherein the sense strand of the siRNA comprises any of SEQ ID NOs 24 to 27, 29 to 32, 34 to 37 and 39 to 42, or a nucleotide sequence having at least 75% sequence identity thereto.
  10. 10. The nucleic acid silencing molecule of any of the preceding claims, wherein the sense strand of the siRNA comprises a nucleotide sequence having at least 80%, at least 90% or at least 95% sequence identity to any of SEQ ID NOs 24 to 27, 29 to 32, 34 to 37 and 39 to 42.
  11. 11. The nucleic acid silencing molecule of any of the preceding claims, wherein: (a) The sense strand of the siRNA comprises SEQ ID NO. 26 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 68 or a nucleotide sequence having at least 75% sequence identity thereto; (b) The sense strand of the siRNA comprises SEQ ID NO. 24 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 66 or a nucleotide sequence having at least 75% sequence identity thereto; (c) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 25 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 67 or at least 75% sequence identity thereto; (d) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO 27 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO 69 or at least 75% sequence identity thereto; (e) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 29 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 71 or at least 75% sequence identity thereto; (f) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 30 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 72 or at least 75% sequence identity thereto; (g) The sense strand of the siRNA comprises SEQ ID NO. 31 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 73 or a nucleotide sequence having at least 75% sequence identity thereto; (h) The sense strand of the siRNA comprises SEQ ID NO. 32 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO. 74 or a nucleotide sequence having at least 75% sequence identity thereto; (i) The sense strand of the siRNA comprises SEQ ID NO 34 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 76 or a nucleotide sequence having at least 75% sequence identity thereto; (j) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 35 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 77 or at least 75% sequence identity thereto; (k) The sense strand of the siRNA comprises SEQ ID NO 36 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 78 or a nucleotide sequence having at least 75% sequence identity thereto; (l) The sense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 37 or at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises the nucleotide sequence of SEQ ID NO. 79 or at least 75% sequence identity thereto; (m) the sense strand of said siRNA comprises a nucleotide sequence of SEQ ID No. 39 or having at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises a nucleotide sequence of SEQ ID No. 81 or having at least 75% sequence identity thereto; (n) the sense strand of said siRNA comprises a nucleotide sequence of SEQ ID No. 40 or having at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises a nucleotide sequence of SEQ ID No. 82 or having at least 75% sequence identity thereto; (o) the sense strand of the siRNA comprises SEQ ID NO 41 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of the siRNA comprises SEQ ID NO 83 or a nucleotide sequence having at least 75% sequence identity thereto, or (P) the sense strand of said siRNA comprises SEQ ID NO. 42 or a nucleotide sequence having at least 75% sequence identity thereto, and the antisense strand of said siRNA comprises SEQ ID NO. 84 or a nucleotide sequence having at least 75% sequence identity thereto.
  12. 12. The nucleic acid silencing molecule of 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 to a corresponding one of SEQ ID NOs 66 to 69, 71 to 74, 76 to 79 and 81 to 84.
  13. 13. The nucleic acid silencing molecule of any of the preceding claims, wherein the nucleic acid silencing molecule is conjugated to one or more non-nucleic acid moieties.
  14. 14. The nucleic acid silencing molecule of any 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 from the 5' end of the antisense strand of the siRNA.
  15. 15. An in vitro method of reducing STAT6 expression in a cell, comprising contacting the cell with the nucleic acid silencing molecule of any of the preceding claims.
  16. 16. A method of treating a disease in a subject, comprising administering to the subject a composition comprising the nucleic acid silencing molecule of any of claims 1 to 14.
  17. 17. The method of claim 16, wherein the disease is cancer.
  18. 18. The method of claim 16, wherein the disease is a disease in which the subject is able to benefit from a reduced or relatively reduced antigen-specific Th2 response.
  19. 19. The method of claim 16 or 18, wherein the disease is an inflammatory disease.
  20. 20. The method of claim 16 or 18, wherein the disease is an allergic disease.

Description

STAT6 siRNA Technical Field The present disclosure relates to nucleic acid silencing molecules comprising or consisting of small interfering RNAs (sirnas) that reduce expression of signal transduction and transcription activator 6 (STAT 6). The disclosure also relates to in vitro methods of reducing STAT6 expression in a cell. The present disclosure further relates to methods of treating a disease in a subject, and related medical uses, comprising administering to the subject a composition comprising the nucleic acid silencing molecule comprising or consisting of an siRNA that reduces STAT6 expression. Background Signal transduction and transcription activators (Signal Transducers and Activators of Transcription, STAT) are a family of transcription factors that contain SH 2. STATs exist in the cytoplasm in inactive form until they are activated by phosphorylation of Janus kinases (JAKs). JAK is associated with the signal transduction chain of the family of erythropoietin in cytokine receptors. When such receptors bind cytokines, they dimerize. This enables JAKs on the signal transduction chain to cross-phosphorylate each other, thereby stimulating their kinase activity. They are then able to recruit STATs by phosphorylating a conserved tyrosine residue at the carboxy terminus. This causes conformational changes in STAT, forming dimers capable of exerting downstream effects. In particular, phosphorylated STAT dimers enter the nucleus and act as transcription factors to initiate expression of certain genes. STAT regulated genes include those related to the growth and differentiation of specific lymphocyte subpopulations. Thus, STAT is activated by cytokines, growth factors, and hormonal signaling. There are seven STATs, namely 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 expression of genes required for Th2 cell development. Th2 helper cells provide T cell helper for humoral immune responses (i.e., antibody-mediated immunity). Their effector cytokines are IL-4, IL-5, IL-9, IL-10, IL-13 and IL-25, and their primary effector cells are eosinophils, basophils, mast cells and B cells, and IL-4/IL-5 CD4T cells. Th2 cells play an important role in immunity against extracellular parasites, bacteria and allergens. However, excessive activation of Th2 cells may lead to inflammation and/or allergic diseases, such as allergic rhinitis, atopic dermatitis and asthma. STAT6 is also highly expressed in multiple types of cancers (including breast, pancreas, endometrial, bowel, skin, lung, prostate, thyroid, and colorectal), and has a key role in initiation of tumorigenesis and malignant transformation. In cancer cells, STAT6 regulates gene expression involved in immune and inflammatory responses, cell survival, tumor proliferation and metastasis. Cancers expressing STAT6 have been shown to be associated with lower survival, increased lymph node metastasis, and adverse changes in inflammatory responses. Thus, mechanisms that reduce STAT6 expression are ideal for treating a range of diseases. Disclosure of Invention The inventors have identified a number of novel and advantageous small interfering RNAs (sirnas) that reduce STAT6 expression. Newly identified STAT6 siRNA can be used to treat diseases caused by STAT6 expression or contributing to adverse clinical effects. For example, STAT6 siRNA of the present disclosure may be used to treat cancer or immune system diseases. For example, STAT6 siRNA of the present disclosure may be used to treat inflammatory diseases. Accordingly, the present disclosure provides a nucleic acid silencing molecule comprising or consisting of a small interfering siRNA that reduces STAT6 expression, wherein the siRNA targets exons 5, 15, 19 or 10 of STAT 6. The present disclosure further provides for: -an in vitro method of reducing STAT6 expression in a cell comprising contacting the cell with a nucleic acid silencing molecule of the present disclosure; a method of treating a disease in a subject comprising administering to the subject a composition comprising a nucleic acid silencing molecule of the disclosure; a composition comprising a nucleic acid silencing molecule of the disclosure for use in a method of treating a disease. Drawings FIG. 1:A) schematic diagrams of siRNA1, B) siRNA1w, C) siRNA1a, D) siRNA1B, E) siRNA1C, and F) siRNA 1D. FIG. 2 is a schematic diagram of A) siRNA2, B) siRNA2a, C) siRNA2B, D) siRNA2C, and E) siRNA 2D. FIG. 3 is a schematic diagram of A) siRNA3, B) siRNA3a, C) siRNA3B, D) siRNA3C, and E) siRNA 3D. FIG. 2 is a schematic diagram of A) siRNA4, B) siRNA4a, C) siRNA4B, D) siRNA4C, and E) siRNA 4D. Description of the sequence Listing SEQ ID NO: 1-the base sequence of the sense strand of siRNA1 (i.e., the unmodified nucleotide sequence). SEQ ID NO:2-siRNA1w sense strand (i.e., unmodified nucleotide sequence). SEQ ID NO:3-siRNA1a sense stra