SPLICE-SWITCHING OLIGONUCLEOTIDES AND METHODS OF USE
The present disclosure provides methods and compositions for the treatment of cancer. In some aspects, the present disclosure provides splice-switching oligonucleotides that downregulate AR or EGFR expression and methods of using these splice-switching oligonucleotides to treat cancer.
1 . A splice-switching oligonucleotide (SSO) comprising a sequence complementary to the region comprising the 5′ splice site of Exon 1 or Exon CE3 of the androgen receptor (AR) and wherein the oligonucleotide is chemically modified.
2 . The splice-switching oligonucleotide of claim 1 , wherein the oligonucleotide comprises a sequence that is complementary to SEQ ID NO: 1.
3 . The splice-switching oligonucleotide of claim 2 , wherein the oligonucleotide comprises SEQ ID NO: 3.
4 . The splice-switching oligonucleotide of claim 1 , wherein the oligonucleotide comprises a sequence that is complementary to SEQ ID NO: 2.
5 . The splice-switching oligonucleotide of claim 4 , wherein the oligonucleotide comprises SEQ ID NO: 4.
6 . The splice-switching oligonucleotide of claim 3 , wherein the oligonucleotide comprising:
(i) a length of at least 18 nucleotides and no more than 100 nucleotides,
(ii) complementarity to a sequence within exon CE3 (SEQ ID NO: 1) of an Androgen receptor (AR) pre-mRNA, and
(iii) a modification selected from the group consisting of a 2′-O-methyl phosphorothioate, a morpholino, a phosphorodiamidate-linked morpholino (PMO), a locked nucleic acid (LNA), a peptide nucleic acid, a 2′-O-(2-methoxy ethyl) nucleotide, a G-clamp or 9-(aminoethoxy)phenoxazine nucleotide, and cytosine analogues that form 4 hydrogen bonds with guanosine.
7 . (canceled)
8 . The splice-switching oligonucleotide of claim 5 , wherein the oligonucleotide comprising:
(i) a length of at least 18 nucleotides and no more than 100 nucleotides,
(ii) complementarity to a sequence within exon 1 (SEQ ID NO: 2) of an Androgen receptor (AR) pre-mRNA, and
(iii) a modification selected from the group consisting of a 2′-O-methyl phosphorothioate, a morpholino, a phosphorodiamidate-linked morpholino (PMO), a locked nucleic acid (LNA), a peptide nucleic acid, a 2′-O-(2-methoxy ethyl) nucleotide, a G-clamp or 9-(aminoethoxy)phenoxazine nucleotide, and cytosine analogues that form 4 hydrogen bonds with guanosine.
9 - 14 . (canceled)
15 . A method of treating a subject suffering from cancer comprising administering to the subject a therapeutically effective amount of a splice-switching oligonucleotide of claim 1 such that the disease is treated.
16 . (canceled)
17 . The method according to claim 15 , wherein the cancer comprises prostate cancer.
18 . The method according to claim 17 , wherein the subject is a human.
19 . The method according to claim 18 , wherein the subject is an African American male suffering from prostate cancer.
20 . The method according to claim 19 , wherein the method further comprises administering to the subject a second cancer therapy.
21 . The method of claim 20 wherein the second cancer therapy is selected from the group consisting of chemotherapy, hormone therapy, androgen therapy, radiation, surgery, vaccine therapy and combinations thereof.
22 . The method of claim 21 , wherein the second cancer therapy is MDV3100 (enzalutamide).
23 . A method of treating a subject suffering from a cancer, comprising administering to the subject a therapeutically effective amount of a splice-switching oligonucleotide of claim 1 and a second cancer therapy in an amount effective to treat the cancer.
24 . The method of claim 23 , wherein the splice-switching oligonucleotide is provided in an effective amount to reduce the expression of AR in the cancer of the subject.
25 . The method of claim 24 , wherein the subject suffers from prostate cancer.
26 . The method of claim 25 , wherein the subject suffers from prostate cancer associated with high expression of AR.
27 . The method of claim 26 , wherein the subject is an African American male suffering from prostate cancer.
28 . The method of claim 27 , wherein the subject is suffering from an aggressive form of the cancer.
29 - 31 . (canceled)
32 . The splice-switching oligonucleotide of claim 1 , wherein the oligonucleotide consists of 18-26 nucleotides.
33 . The splice-switching oligonucleotide of claim 1 , wherein the modified splice-switching oligonucleotide comprises a 2′-O-Me phosphorothioate backbone.
34 . The splice-switching oligonucleotide of claim 1 , which is a morpholino oligonucleotide.
35 . The splice-switching oligonucleotide of claim 1 , wherein the modified splice-switching oligonucleotide comprises a 2′-O-(2-methoxyethyl) backbone.
36 . The splice-switching oligonucleotide of claim 1 , wherein the SSO has a length of at least 18 nucleotides and no more than 50 nucleotides.
37 . The splice-switching oligonucleotide of claim 1 , wherein the SSO has a length of at least 18 nucleotides and no more than 40 nucleotides.
38 . The splice-switching oligonucleotide of claim 1 , wherein the SSO has a length of at least 18 nucleotides and no more than 30 nucleotides.
39 . The splice-switching oligonucleotide of claim 1 , wherein the SSO has a length of 18 nucleotides and comprises a phosphorodiamidate-linked morpholino (PMO) modification.
40 . The splice-switching oligonucleotide of claim 1 , wherein the SSO has a length of 30 nucleotides and comprises a phosphorodiamidate-linked morpholino (PMO) modification.
41 . The splice-switching oligonucleotide of claim 38 , wherein the SSO comprises a phosphorodiamidate-linked morpholino (PMO) modification.