Claims for Patent: 11,401,517

Summary for Patent: 11,401,517

Title:	Modified double-stranded RNA agents
Abstract:	One aspect of the present invention relates to double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene. The sense strand of the dsRNA agent comprises at least one thermally destabilizing nucleotide, and at least one said thermally destabilizing nucleotide occurring at a site opposite to the seed region (positions 2-8) of the antisense strand; and the antisense strand of the dsRNA agent comprises at least two modified nucleotides that provide the nucleotide a steric bulk that is less than or equal to the steric bulk of a 2'-OMe modification, wherein said modified nucleotides are separated by 11 nucleotides in length. Other aspects of the invention relates to pharmaceutical compositions comprising these dsRNA agents suitable for therapeutic use, and methods of inhibiting the expression of a target gene by administering these dsRNA agents, e.g., for the treatment of various disease conditions.
Inventor(s):	Maier; Martin (Cambridge, MA), Foster; Don (Cambridge, MA), Milstein; Stuart (Cambridge, MA), Kuchimanchi; Satya (Cambridge, MA), Jadhav; Vasant (Cambridge, MA), Rajeev; Kallanthottathil (Cambridge, MA), Manoharan; Muthiah (Cambridge, MA), Parmar; Rubina (Cambridge, MA)
Assignee:	ALNYLAM PHARMACEUTICALS, INC. (Cambridge, MA)
Application Number:	16/693,683
Patent Claims:	1. A double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene, comprising a sense strand sequence and an antisense strand sequence complementary to at least one portion of a mRNA of the target gene, each of the sense and antisense strands having 19-25 nucleotides in length, wherein the sense strand is represented by formula (Is): ##STR00076## wherein: B1, B2, and B3 each independently represent a nucleotide containing a modification selected from the group consisting of 2'-Oalkyl, 2'-substituted alkoxy, 2'-substituted alkyl, 2'-halo, ENA, and BNA/LNA; C1 is a thermally destabilizing nucleotide, selected from the group consisting of i) a nucleotide that forms a mismatch pair with the opposing nucleotide in the antisense strand, ii) a nucleotide having an abasic modification, and iii) a nucleotide having a sugar modification, and placed at a site opposite to the seed region (positions 2-8) of the antisense strand; T1 represents a nucleotide comprising a 2'-F modification; n.sup.1 or n.sup.3 is independently 4 to 15 nucleotides in length; n.sup.5 is 1-6 nucleotide(s) in length; n.sup.2 is 3; n.sup.4 is 0-3 nucleotide(s) in length; and wherein the sense strand has 2'-F modifications, and wherein the 2'-F modifications on the sense strand consist of four, and only four, 2'-F modifications, wherein the four 2'-F modifications are at positions 7 and 9-11 from the 5'-end ofthe sense strand. 2. The dsRNA agent of claim 1, wherein n.sup.4 is 1. 3. The dsRNA agent of claim 2, wherein C1 is at position 14-17 of the 5'-end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n.sup.4 is 1. 4. The dsRNA agent of claim 1, wherein C1 has thermally destabilizing modification selected from the group consisting of abasic modification selected from the group consisting of: ##STR00077## and sugar modification selected from the group consisting of: ##STR00078## wherein B is a modified or unmodified nucleobase, R.sup.1 and R.sup.2 independently are H, halogen, OR.sub.3, or alkyl; and R.sub.3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar. 5. The dsRNA agent of claim 1, wherein B1, B2, and B3 each contain 2'-OMe modifications. 6. The dsRNA agent of claim 1, wherein the sense strand comprises one block of two phosphorothioate or methylphosphonate internucleotide linkages. 7. The dsRNA agent of claim 1, wherein the dsRNA agent has a 3' and/or 5' overhang(s) of 1-10 nucleotides in length. 8. The dsRNA agent of claim 1, further comprising at least one ASGPR ligand attached to the 3' end of the sense strand. 9. The dsRNA agent of claim 8, wherein the ASGPR ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 10. The dsRNA agent of claim 9, wherein the ASGPR ligand is: ##STR00079## 11. The dsRNA agent of claim 1, wherein formula (Is) further comprises a 2'-deoxythymidine linked via a phosphorodithioate (PS.sub.2) linkage or a 5'-vinyl phosphonate (VP) at the 5'-end of the sense or antisense strand. 12. A pharmaceutical composition comprising the dsRNA agent according to claim 1 in combination with a pharmaceutically acceptable carrier or excipient. 13. A method for inhibiting the expression of a target gene comprising the step of administering the dsRNA agent according to claim 1, in an amount sufficient to inhibit expression of the target gene. 14. The method of claim 13, wherein the dsRNA agent is administered through subcutaneous or intravenous administration. 15. A method for delivering polynucleotide to specific target in a subject by administering the dsRNA agent according to claim 1. 16. The method of claim 15, wherein said administering step is carried out by an administration means comprising intramuscular, intrabronchial, intrapleural, intraperitoneal, intraarterial, lymphatic, intravenous, subcutaneous, cerebrospinal, or combinations thereof. 17. A method for delivering a polynucleotide to specific target of a subject, the method comprising: delivering a dsRNA agent according to claim 1 by subcutaneous administration into the subject, such that the polynucleotide is delivered into specific target of the subject. 18. The dsRNA agent of claim 1, wherein n.sup.4 is 0. 19. The dsRNA agent of claim 1, wherein each B1, B2, and B3 is independently 2'-OMe or 2'-F. 20. The dsRNA agent of claim 1, wherein C1 is at position 14-17 of the 5'-end of the sense strand, n.sup.4 is 1, and C1 is a glycol nucleic acid (GNA). 21. The dsRNA agent of claim 1, wherein C1 is at position 14-17 of the 5'-end of the sense strand, n.sup.4 is 1, C1 is a glycol nucleic acid (GNA), and each B1, B2, and B3 is independently 2'-OMe or 2'-F. 22. The dsRNA agent of claim 1, wherein n.sup.4 is 0, and each B1, B2, and B3 is independently 2'-OMe or 2'-F. 23. The dsRNA agent of claim 1, wherein each B1 is independently 2'-OMe or 2'-F, and each B2 and B3 is 2'-OMe. 24. The dsRNA agent of claim 1, wherein each B2 and B3 is 2'-OMe.