Stabilized STAT3 decoy oligonucleotides and uses therefor
The present invention is based, at least in part, on novel, unimolecular STAT3 oligonucleotide decoys exhibiting increased in vivo stability as compared to previously known decoys which are effective in inhibiting STAT3 when administered systemically. The invention is also based on pharmaceutical compositions comprising these unimolecular decoys, and methods for using these decoys in the treatment of cancer.
1. A cyclic double-stranded STAT3 oligonucleotide decoy, wherein (i) the decoy comprises a double-stranded oligonucleotide, or an analog thereof, having the sequence
5′-(N 6 -) n CAN 1 TTCN 2 CN 3 TN 4 AN 5 TC-(N 7 -) m -3′ (SEQ ID NO:1), wherein N 3 is G; N 1 , N 2 , N 4 and N 5 are A, T, G or C; one, two, three or all of the following conditions are met: N 1 is T; N 2 is C; N 4 is A and N 5 is A; and N 6 and N 7 are A, T, G or C and n and m are independently 0-50; (ii) the two strands are joined at the ends by chemical linkers; (iii) the decoy binds to STAT3 protein under physiologic conditions and interferes with STAT3 binding to its target sequence; and (iv) the decoy has a serum half-life of greater than about 4 hours.
2. The STAT3 oligonucleotide decoy of claim 1 , wherein the oligonucleotide comprises the sequence 5′-CATTTCCCGTAAATC-3′ (SEQ ID NO:30).
3. The STAT3 oligonucleotide decoy of claim 1 , wherein N 2 is a pyrimidine.
4. The STAT3 oligonucleotide decoy of claim 1 , wherein at least two of the following are met: N 1 is T; N 2 is C; N 4 is A; and N 5 is A.
5. The STAT3 oligonucleotide decoy of claim 1 , wherein the chemical linker is a chemical structure comprising a carbon spacer.
6. The STAT3 oligonucleotide decoy of claim 5 , wherein the carbon spacer has 18 carbons.
7. The STAT3 oligonucleotide decoy of claim 5 , wherein the carbon spacer comprises an ethylene glycol spacer, optionally wherein the ethylene glycol is a hexaethylene glycol.
8. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a cyclic double-stranded STAT3 oligonucleotide decoy, wherein (i) the decoy comprises a double-stranded oligonucleotide, or an analog thereof, having the sequence 5′-(N 6 -) n CA N 1 TTCN 2 C N 3 T N 4 A N 5 TC-(N 7 -) m -3′ (SEQ ID NO:1), wherein N 3 is G; N 1 , N 2 , N 4 and N 5 are A, T, G or C; one, two, three or all of the following conditions are met: N 1 is T; N 2 is C; N 4 is A and N 5 is A; and N 6 and N 7 are A, T, G or C and n and m are independently 0-50; (ii) the two strands are joined at the ends by chemical linkers; (iii) the decoy binds to STAT3 protein under physiologic conditions and interferes with STAT3 binding to its target sequence; and (iv) the decoy has a serum half-life of greater than about 4 hours.
9. The pharmaceutical composition of claim 8 , further comprising an anticancer agent.
10. The composition of claim 8 , formulated as a parenteral dosage form or an intravenous dosage form.
11. The composition of claim 8 , wherein the decoy is contained within a microbubble.
12. The composition of claim 8 , wherein the decoy is associated with a peptide transduction domain, optionally wherein the peptide transduction domain is TAT.
13. A method of inhibiting growth of a cancer in which STAT3 is activated in a patient, comprising administering to the patient an amount of a cyclic double-stranded STAT3 oligonucleotide decoy effective to inhibit growth of a cancer in a patient, thereby inhibiting growth of the cancer in the patient; wherein the oligonucleotide decoy (i) comprises a double-stranded oligonucleotide having the sequence 5′-(N 6 -) n CAN 1 TTCN 2 CN 3 TN 4 AN 5 TC-(N 7 -) m -3′ (SEQ ID NO: 1), wherein N 3 is G; N 1 , N 2 , N 4 and N 5 are A, T, G or C; one, two, three or all of the following conditions are met: N 1 is T; N 2 is C; N 4 is A and N 5 is A; and N 6 and N 7 are A, T, or C and n and m are independently 0-50; (ii) the two strands are joined at the ends by two chemical linkers; (iii) the decoy binds to STAT3 protein under physiologic conditions and interferes with STAT3 binding to its target sequence; and (iv) the decoy has a serum half-life of greater than about 4 hours.
14. The method of claim 13 , comprising administering to the patient a second anticancer therapy comprising one or both of a radiation therapy and treating the patient with an anticancer agent.
15. The methods of claim 14 , wherein the anticancer therapy comprises an epidermal growth factor receptor (EGFR) antagonist, optionally wherein the EGFR antagonist is cetuximab.
16. The method of claim 13 , wherein the cancer is selected from the group consisting of multiple myeloma, HTLV-1 dependent leukemia, acute myelogenous leukemia, large granular lymphocyte leukemia, lymphoma, EBV-related Burkitt's lymphoma, mycosis fungoides, cutaneous T-cell lymphoma, non-Hodgkins lymphoma, anaplastic large-cell lymphoma, breast cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, prostate cancer and squamous cell carcinoma, optionally wherein the squamous cell carcinoma is squamous cell carcinoma of the head and neck.
17. The method of claim 13 , wherein the STAT3 oligonucleotide decoy comprises the sequence 5′CATTTCCCGTAAATC-3′ (SEQ ID NO:30).
18. The method of claim 13 , wherein, in the STAT3 oligonucleotide decoy, N 2 is a pyrimidine or at least two of the following are met: N 1 is T; N 2 is C; N 4 is A; and N 5 is A.
19. The method of claim 13 , wherein, in the STAT3 oligonucleotide decoy, the chemical linker is a chemical structure comprising a carbon spacer.
20. The method of claim 19 , wherein the carbon spacer comprises an ethylene glycol spacer, optionally wherein the ethylene glycol is a hexaethylene glycol.