Oligonucleotide modulators of the toll-like receptor pathway
Disclosed herein are double stranded nucleic acid molecules and pharmaceutical compositions comprising same useful in the treatment of, inter alia, acute and chronic inflammation, neuropathic pain, primary graft dysfunction (PGD) after lung transplantation in a subject in need thereof. The compounds are preferably chemically synthesized and modified dsRNA compounds, which down regulate or inhibit expression of Toll like receptor 4.
1. A double stranded nucleic acid molecule comprising a sense strand and an antisense strand described as the sequence pair set forth as TLR4_14 (SEQ ID NOS: 20624 and 20633, respectively).
2. The double stranded nucleic acid molecule of claim 1 having the following structure:
(A2)
5′ N1-(N)x-Z 3′ (antisense strand)
3′ Z′-N2-(N′)y-z″ 5′ (sense strand)
wherein each N2, N and N′ is an unmodified or modified ribonucleotide, or an unconventional moiety;
wherein each of (N)x and (N′)y is an oligonucleotide in which each consecutive N or N′ is joined to the adjacent N or N′ by a covalent bond;
wherein each of x and y is independently an integer between 17 and 24;
wherein the sequence of (N′)y is complementary to the sequence of (N)x and (N)x is complementary to a consecutive sequence in a target RNA set forth in any one of SEQ ID NO:2-4 (TLR4 mRNA);
wherein N1 is an unmodified or modified ribonucleotide covalently bound to (N)x and mismatched to the target RNA;
wherein z″ may be present or absent, but if present is a capping moiety covalently attached at the 5′ terminus of N2-(N′)y;
wherein each of Z and Z′ is independently present or absent, but if present is independently 1-5 consecutive nucleotides, consecutive non-nucleotide moieties or a combination thereof covalently attached at the 3′ terminus of the strand in which it is present; and
wherein the sequence of N1-(N)x is set forth in SEQ ID NO: 20633.
3. The double stranded nucleic acid molecule of claim 2 , wherein at least one of N1, N2, N or N′ comprises a modified nucleotide or an unconventional moiety.
4. The double stranded nucleic acid molecule of claim 3 , wherein N1-(N)x comprises at least one pyrimidine ribonucleotide and wherein at least one of the pyrimidine ribonucleotides in N1-(N)x comprises a 2′ sugar modified pyrimidine ribonucleotide.
5. The double stranded nucleic acid molecule of claim 4 , wherein the 2′ sugar modified ribonucleotide comprises a 2′-OMe sugar modified ribonucleotide.
6. A method for the treatment of a subject in need of treatment for a disease or disorder or symptom or condition associated with the expression of a target gene comprising administering to the subject an amount of a double stranded nucleic acid molecule of claim 1 , in an amount effective to down regulate gene expression, wherein the gene encodes a RNA having a polynucleotide sequence as set forth in any one of SEQ ID NO:2-4 (TLR4 mRNA).
7. The method of claim 6 , wherein the disease or injury is selected from the group consisting of: chronic or acute aseptic inflammation, neuropathic pain, primary graft failure, ischemia-reperfusion injury, reperfusion injury, reperfusion edema, allograft dysfunction, pulmonary reimplantation response and primary graft dysfunction (PGD) in organ transplantation.
8. The method of claim 7 , wherein the disease or injury comprises primary graft dysfunction (PGD) in organ transplantation.
9. The method of claim 8 , wherein said organ transplantation comprises lung transplantation.
10. A pharmaceutical composition comprising a double stranded nucleic acid molecule of claim 1 in an amount effective to inhibit gene expression, and a pharmaceutically acceptable carrier wherein the gene encodes a RNA having a polynucleotide sequence set forth in any one of SEQ ID NO:2-4 (TLR4 mRNA).