Double-stranded ribonucleic acids with rugged physico-chemical structure and highly specific biologic activity
The invention relates to our discovery of a novel double-stranded ribonucleic acid (dsRNA) having specific biological activities, which includes acting as a selective agonist for activation of the Toll-like receptor 3. Its “rugged” molecular structure as measured by physico-chemical techniques is resistant to molecular unfolding (i.e., denaturation). This structure appears to be responsible for increased efficacy of dsRNA in therapeutic applications and improved biological activity (e.g., used as an immunoregulatory agent). Medicaments, processes for their manufacture, and methods for their use are provided herein.
1. An isolated double-stranded ribonucleic acid (dsRNA) enzymatically active under thermal stress comprising:
each strand with a molecular weight of about 250 Kda to about 320 Kda, a single strand comprised of poly(ribocytosinic 4-29 uracilic acid) and an opposite strand comprised of poly(riboinosinic acid), wherein the two strands do not base pair the position of the uracil base, wherein the two strands base pair the position of the cytosine base, and wherein said strands are partially hybridized.
2. An isolated double-stranded ribonucleic acid (dsRNA) enzymatically active under thermal stress comprising:
each strand of a length from about 380 bases to about 450 bases, a single strand comprised of poly(ribocytosinic 4-29 uracilic acid) and an opposite strand comprised of poly(riboinosinic acid), wherein the two strands do not base pair the position of the uracil base, wherein the two strands base pair the position of the cytosine base, and wherein said strands are partially hybridized.
3. An isolated double-stranded ribonucleic acid (dsRNA) enzymatically active under thermal stress comprising:
each strand with about 30 to 38 helical turns of duplexed RNA strands, a single strand comprised of poly(ribocytosinic 4-29 uracilic acid) and an opposite strand comprised of poly(riboinosinic acid), wherein the two strands do not base pair the position of the uracil base, wherein the two strands base pair the position of the cytosine base, and wherein said strands are partially hybridized.
4. A composition comprising one or more different dsRNAs defined in claim 1 .
5. The dsRNA of claim 1 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 11 - 14 U) n .
6. The dsRNA of claim 5 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 12 U) n .
7. A composition comprising one or more different dsRNAs as defined in claim 2 .
8. The dsRNA of claim 2 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 11 - 14 U) n .
9. The dsRNA of claim 8 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 12 U) n .
10. A composition comprising one or more different dsRNAs defined in claim 3 .
11. The dsRNA of claim 3 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 11 - 14 U) n .
12. The dsRNA of claim 11 , wherein said dsRNA is comprised of ribo(I n ).ribo(C 12 U) n .
13. The dsRNA of claim 1 , wherein the double-stranded ribonucleic acid is a purified double-stranded ribonucleic acid.