Compounds and compositions including phosphorothioated oligodeoxynucleotide, and methods of use thereof
The present disclosure relates to an isolated compound including a phosphorothioated oligodeoxynucleotide (ODN) sequence conjugated to a short-activating RNA (saRNA) or an antisense oligonucleotide sequence (ASO), compositions of such a compound, and method of treatment of cancer and autoimmune diseases (with or without stimulating immune response), method of immune stimulation, method of activating CEBPA, and method of reducing activity of STAT transcription factor, by one of the disclosed compounds or compositions.
1. A method of modulating expression of a target gene in a subject, the method comprising administering to the subject a phosphorothioated oligodeoxynucleotide (ODN) conjugate, wherein the conjugate comprises a phosphorothioated ODN sequence conjugated to: (i) a short-activating RNA (saRNA) of CCAAT/enhancer-binding protein-α (C/EBPα), p21, or p53 or (ii) an antisense oligonucleotide (ASO) sequence of Signal Transducer and Activator of Transcription (STAT), wherein the ASO comprises a single-stranded DNA substituent.
2. The method of claim 1 , wherein said antisense oligonucleotide sequence is an anti-STAT1 oligonucleotide sequence, anti-STAT2 oligonucleotide sequence, anti-STAT3 oligonucleotide sequence, anti-STAT4 oligonucleotide sequence, anti-STAT5A oligonucleotide sequence, anti-STAT5B oligonucleotide sequence, or anti-STAT6 oligonucleotide sequence.
3. The method of claim 1 , wherein the conjugate comprises a linker between the phosphorothioated ODN sequence and the short-activating RNA (saRNA) or the ASO.
4. The method of claim 3 , wherein the linker is a substituted or unsubstituted C 1 -C 40 alkylene, substituted or unsubstituted 2 to 40 membered heteroalkylene, substituted or unsubstituted C 3 -C 8 cycloalkylene, substituted or unsubstituted 3 to 8 membered heterocycloalkylene, substituted or unsubstituted C 6 -C 10 arylene, or substituted or unsubstituted 5 to 10 membered heteroarylene.
5. The method of claim 3 , wherein the linker is a substituted 2 to 40 membered heteroalkylene.
6. The method of claim 1 , wherein the saRNA or the ASO comprises a chemical modification selected for the group consisting of a 2′ O-Methyl, 2′-deoxy-2′fluoro, 2′-deoxy, a universal base, 5-C-methyl, an inverted deoxy abasic residue incorporation, and a locked nucleic acid.
7. The method of claim 1 , further comprising administering a second therapeutic agent to the subject, wherein the second therapeutic agent is selected from the group consisting of: anti-tumor or anti-cancer agent, cytotoxic agent, cytostatic agent, anti-inflammatory agent, analgesic, anti-infective agent, growth inhibitory agent, immunogenic agent, immunomodulatory agent, and chemokine.
8. The method of claim 1 , wherein the conjugate is administered in an amount effective to treat cancer in the subject, while simultaneously stimulating an immune response.
9. The method of claim 1 , wherein the conjugate comprises: (i) a saRNA of CEBPA, p21, or p53 conjugated to one of phosphorothioated oligodeoxynucleotides (ODN) of SEQ ID NOs: 7-18 and 98-101, or (ii) a STAT ASO of one of SEQ ID NOs: 31-42 and 110-113 conjugated to one of phosphorothioated oligodeoxynucleotides (ODN) of SEQ ID NO: 7-18 and 98-101.
10. The method of claim 1 , wherein the conjugate is administered in an amount effective to treat cancer in the subject without simultaneously stimulating an immune response.
11. The method of claim 1 , wherein the conjugate comprises saRNA of CEBPA, p21, or p53, or one of STAT ASO of SEQ ID NOs: 31-42 and 110-113, conjugated to one of phosphorothioated oligodeoxynucleotides (ODN) of SEQ ID NO: 29-30.
12. The method of claim 1 , wherein the conjugate is administered in an amount effective to treat an autoimmune disease in the subject without simultaneously stimulating an immune response.
13. The method of claim 1 , wherein the conjugate is administered to the subject by intravenous, parenteral, subcutaneous, intramuscular, transdermal, intraperitoneal, intranasal, aerosol, oral, or topical administration.
14. The method of one of claim 13 , wherein about 0.001 mg/kg to about 100 mg/kg of said conjugate is administered to said subject.
15. The method of claim 1 , wherein the conjugate is administered in an amount effective to stimulate an immune response in the subject.
16. The method of claim 15 , wherein said immune response comprises:
(a) maturation, differentiation, or proliferation of natural killer cells, T cells, B cells or myeloid cells;
(b) an increase in T H 1-type immune response;
(c) recruiting dendritic cells and CD8+ T cells into an organ of said subject;
(d) expanding a population of antigen-presenting cells in said subject; or
(e) suppressing proliferation of cancer cells in said subject.
17. The method of claim 1 , wherein (i) the conjugate comprises the saRNA of CEBPA, and (ii) the conjugate is administered in an amount effective to enhance C/EBPα expression in a cell.
18. The method of claim 1 , wherein the conjugate is administered in an amount effective to inhibit cell growth.
19. The method of claim 1 , wherein (i) the conjugate comprises the STAT ASO, and (ii) the conjugate is administered in an amount effective to reduce activity of a STAT transcription factor in a cell.
20. The method of claim 17 , wherein said cell is a cancer cell.