Dual molecular delivery of oligonucleotides and peptide containing conjugates

Patent metadata

Field	Value
Publication number	US-10532068-B2
Application number	US-201815974135-A
Country	US
Kind code	B2
Filing date	May 8, 2018
Priority date	Nov 6, 2013
Publication date	Jan 14, 2020
Grant date	Jan 14, 2020

Abstract

Disclosed herein is a method for inhibiting expression of a gene of a subject comprising administering (1) a composition comprising R-(L)a-(G)b; wherein R is an oligonucleotide selected from the group consisting of DNA, RNA, siRNA, and microRNA; L is a linker and each occurrence of L is independently selected from Table 3; G is a targeting ligand and each occurrence of G is independently selected from Table 4; each of a and b is independently 0, 1, 2, 3 or 4; and (2) a composition comprising (P)c-(L)d-(G)e; wherein P is a peptide and each occurrence of P is independently selected from Table 2; L is a linker and each occurrence of L is independently selected from Table 3; G is a targeting ligand and each occurrence of G is independently selected from Table 4; d is 0, 1, 2, 3, 4, 5 or 6; and each of c and e is independently 1, 2, 3, 4, 5 or 6. Compositions in (1) and (2) can be co-administered or sequentially administered.

First claim

What is claimed is:

1. A method for inhibiting expression of a gene of a subject comprising administering:
(1) a composition comprising R-(L) a -(G) b to the subject; wherein:
R is an oligonucleotide selected from the group consisting of DNA, RNA, siRNA, and microRNA;
L is a linker and each occurrence of L is independently selected from Table 3:

G is a targeting ligand and each occurrence of G is independently selected from Table 4:

each of a and b is independently 0, 1, 2, or 3; and
(2) a composition comprising (P) c -(L) d -(G) e to the subject; wherein:
P is a peptide and each occurrence of P is independently selected from SEQ ID NOS: 1-1802;
L is a linker and each occurrence of L is independently selected from Table 3;
G is a targeting ligand and each occurrence of G is independently selected from Table 4;
d is 0, 1, 2, 3, or 4; and
each of c and e is independently 1, 2, 3, or 4.

2. The method of claim 1 , wherein R is a double stranded siRNA or single stranded siRNA.

3. The method of claim 1 , wherein each occurrence of L is independently selected from Table 3a:

4. The method of claim 1 , wherein each occurrence of G is independently selected from Table 4a:

5. The method of claim 4 , wherein G is a ligand of the following formula:

6. The method of claim 1 , wherein:
each of a and b is independently 0, 1 or 2;
c is 1 or 2; and
each of d and e is independently 1 or 2.

7. The method of claim 1 , comprising administering:
(1) a composition comprising R-(L) a -(G) b to the subject; wherein:
R is an siRNA;
L is a linker and each occurrence of L is independently selected from Table 3a:

G is a targeting ligand and each occurrence of G is independently selected from Table 4a:

each of a and b is independently 0, 1 or 2; and
(2) a composition comprising (P) c -(L) d -(G) e to the subject; wherein:
P is a peptide and each occurrence of P is independently selected from SEQ ID NOS: 1697, 1701-1713, 1715-1734, 1738, 1740, 1741, 1744-1749, 1751-1753, 1755, 1756, 1758-1786, 1788, and 1793-1802;
L is a linker and each occurrence of L is independently selected from Table 3a;
G is a targeting ligand and each occurrence of G is independently selected from Table 4a; and
each of c, d and e is independently 1 or 2.

8. The method of claim 7 , wherein:
L of R-(L) a -(G) b is selected from

 wherein m is an integer selected from 0 to 10;
G of R-(L) a -(G) b is:

 and
(P) c -(L) d -(G) e of composition (2) is:

wherein P is a peptide selected from SEQ ID NOS: 1697, 1701-1713, 1715-1734, 1738, 1740, 1741, 1744-1749, 1751-1753, 1755, 1756, 1758-1786, 1788, and 1793-1802; and
wherein L of composition (2) is selected from:

 wherein m is an integer selected from 0 to 10,

9. The method of claim 8 , wherein each L of compositions (1) and (2) is independently

10. The method of claim 1 , wherein:
the composition comprising R-(L) a -(G) b and the composition comprising (P) c -(L) d -(G) e are co-administered at the same time.

11. The method of claim 1 , wherein:
the composition comprising R-(L) a -(G) b and the composition comprising (P) c -(L) d -(G) e are sequentially administered about 0.1 to 1 hour apart.

12. The method of claim 1 , wherein the oligonucleotide is administered at a dose of 0.1 to 5 mpk; and the peptide is administered at a dose of 1 to 100 mpk.

13. A composition for dual molecular delivery of an oligonucleotide and a peptide conjugate comprising:
(1) R-(L) a -(G) b ; and
(2) (P) c -(L) d -(G) e ; wherein:
R is an oligonucleotide selected from the group consisting of DNA, RNA, siRNA, and microRNA;
P is a peptide and each occurrence of P is independently selected from SEQ ID NOS: 1697, 1701-1713, 1715-1734, 1738, 1740, 1741, 1744-1749, 1751-1753, 1755, 1756, 1758-1786, 1788, and 1793-1802;
L is a linker and each occurrence of L is independently selected from Table 3 of claim 1 ;
G is a targeting ligand and each occurrence of G is independently selected from Table 4 of claim 1 ;
each of a and b is independently 0, 1, 2, or 3; and
each of c, d and e is independently 1, 2, 3, 4, or 5.

14. The composition of claim 13 , wherein Y comprises a ligand of the following formula:

15. The composition of claim 13 , wherein each of a and b is independently 0 or 1;
c is 1; and
each of d and e is an 1.

16. The composition of claim 13 , wherein:
(1) G of R-(L) a -(G) b is:

 and
each of a and b is 1; and
(2) (P) c -(L) d -(G) e is:

 and
wherein each L of compositions (1) and (2) is independently selected from:

17. The composition of claim 13 , wherein R-(L) a -(G) b further comprises a lipid and/or solubilizing agent.

18. The composition of claim 13 , wherein:
the oligonucleotide is a double stranded siRNA; and
G is attached to the guide strand or the passenger strand of the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions.

Assignees

• Merck Sharp & Dohme

Inventors

• Colletti Steven L
• Tucker Thomas J
• Tellers David M
• Kim Boyoung
• Burke Rob
• Calati Kathleen B
• Stanton Matthew G
• Parmar Rubina G
• Aaronson Jeffrey G
• Wang Weimin

Classifications

• A61K38/10

  Peptides having 12 to 20 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title

• C07K2319/10

  containing a tag for extracellular membrane crossing, e.g. TAT or VP22 · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title