Skeletal Muscle Delivery Platforms and Methods of Use
The present disclosure relates to delivery platforms that specifically and efficiently direct payloads to skeletal muscle cells in a subject, in vivo. The delivery platforms disclosed herein include targeting ligands (such as compounds that have affinity for integrins, including alpha-v-beta-6) and pharmacokinetic/pharmacodynamic (PK/PD) modulators, to facilitate the delivery of payloads to cells, including to skeletal muscle cells. Suitable payloads for use in the delivery platforms disclosed herein include RNAi agents, which can be linked or conjugated to the delivery platforms, and when delivered in vivo, provide for the inhibition of gene expression in skeletal muscle cells. Pharmaceutical compositions that include the skeletal muscle cell delivery platform are also described, as well as methods of use for the treatment of various diseases and disorders where delivery of a therapeutic payload to a skeletal muscle cell is desirable.
1 . A delivery platform for inhibiting expression of a gene expressed in skeletal muscle cells comprising:
(a) An RNAi agent comprising:
(i) An antisense strand comprising 17-49 nucleotides wherein at least 15 nucleotides are complementary to the mRNA sequence of a gene that is expressed in skeletal muscle cells
(ii) A sense strand that is 16-49 nucleotides in length that is at least partially complementary to the antisense strand;
(b) A targeting ligand with affinity for a receptor present on the surface of a skeletal muscle cell; and
(c) A PK/PD modulator;
wherein the RNAi agent is covalently linked to the targeting ligand and to the PK/PD modulator.
2 . The delivery platform of claim 1 , wherein the targeting ligand has affinity for an integrin receptor.
3 . The delivery platform of claim 1 , wherein the targeting ligand has affinity for the αvβ6 integrin receptor.
4 . The delivery platform of claim 1 , wherein the targeting ligand is of the formula:
or a pharmaceutically acceptable salt thereof,
wherein,
n is an integer from 0 to 7;
J is C—H or N;
Z is OR 13 , N(R 13 ) 2 or SR 13 ;
R 1 is H, optionally substituted C 1 -C 6 alkyl, OH, COOH, CON(R 5 ) 2 , OR 6 , or R 1 comprises a cargo molecule, wherein each R 5 is independently H or C 1 -C 6 alkyl, and R 6 is H or C 1 -C 6 alkyl;
R 2 , R 1′ and R 2 are each independently H, halo, optionally substituted cycloalkylene, optionally substituted arylene, optionally substituted heterocycloalkylene, or optionally substituted heteroarylene, or R 2 , R 1′ and R 2 may comprise a cargo molecule;
R 10 is H or optionally substituted alkyl;
R 11 is H or optionally substituted alkyl, or R 11 and R 1 together with the atoms to which they are attached form an optionally substituted heterocycle;
R 12 is H or optionally substituted alkyl;
each R 13 is independently H, optionally substituted alkyl, or R 13 comprises a cargo molecule;
R 14 is optionally substituted alkyl; and
wherein at least one of R 1 , R 2 , R 13 , R P1 and R 2 comprises the antisense strand.
5 . The delivery platform of claim 4 , wherein the targeting ligand is selected from the group consisting of:
or a pharmaceutically acceptable salt thereof, wherein indicates the point of connection to the RNAi agent.
6 . (canceled)
7 . The delivery platform of claim 1 , wherein the targeting ligand has the formula:
or a pharmaceutically acceptable salt thereof, wherein
R 1 is optionally substituted alkyl, optionally substituted alkoxy, or
wherein R 11 and R 12 are each independently optionally substituted alkyl;
R 2 is H or optionally substituted alkyl;
R 3 is H or optionally substituted alkyl;
R 4 is H or optionally substituted alkyl;
R 5 is H or optionally substituted alkyl;
R 6 is selected from the group consisting of H, optionally substituted alkyl, optionally substituted alkoxy, halo, optionally substituted amino;
Q is optionally substituted aryl or optionally substituted alkylene;
X is O, CR 8 R 9 , NW;
wherein R 8 is selected from H, optionally substituted alkyl, or R 8 is taken together with Rx or Ry to form a 4-, 5-, 6-, 7-, 8- or 9-membered ring, and R 9 is H or optionally substituted alkyl;
Rx and Ry are each independently H, optionally substituted alkyl, or Rx and Ry may be taken together to form a double bond with R 10 , wherein R 10 is H, optionally substituted alkyl, or R 10 may be taken together with X and the atoms to which it is attached to form a 4-, 5-, 6-, 7-, 8, or 9-membered ring;
wherein at least one of R 1 , R 2 , R 6 , R 11 , R 12 , Rx and Ry comprise a cargo molecule; and
wherein when Q is optionally substituted alkyl and the length of the optionally substituted alkyl chain represented by Q is 3 carbons, then R 1 is
8 . The delivery platform of claim 7 , wherein the targeting ligand comprises a structure selected from the group consisting of:
Compound
Number
Formula
40b
41b
42b
43b
44b
45b
46b
47b
48b
49b
50b
51b
52b
53b
54b
55b
56b
57b
58b
59b
60b
or a pharmaceutically acceptable salt thereof, and wherein indicates the point of connection to a cargo molecule.
9 . (canceled)
10 . The delivery platform of claim 2 , wherein the PK/PD modulator comprises at least one polyethylene glycol (PEG) unit.
11 . (canceled)
12 . The delivery platform of claim 10 , wherein the PK/PD modulator is selected from the group consisting of:
wherein indicates the point of attachment to the RNAi agent.
13 . The delivery platform of claim 2 , wherein the PK/PD modulator has the formula:
or a pharmaceutically acceptable salt thereof, wherein
L A is a bond or a bivalent moiety connecting Z to the RNAi agent;
Z is CH, phenyl, or N;
L 1 and L 2 are each independently linkers comprising at least about 5 PEG units;
X and Y are each independently lipids comprising from about 10 to about 50 carbon atoms; and
indicates a point of connection to the RNAi agent.
14 - 18 . (canceled)
19 . The delivery platform of claim 13 , wherein at least one of X and Y is an unsaturated lipid.
20 - 23 . (canceled)
24 . The delivery platform of claim 13 , wherein at least one of X and Y is cholesteryl.
25 . The delivery platform of claim 13 , wherein at least one of X and Y is selected from the group consisting of:
Name
Structure
Lipid 1
Lipid 2
Lipid 3
Lipid 4
Lipid 5
Lipid 6
Lipid 7
Lipid 8
Lipid 9
Lipid 10
Lipid 11
Lipid 12
Lipid 14
Lipid 15
Lipid 16
Lipid 17
Lipid 18
Lipid 19
Lipid 20
Lipid 21
Lipid 22
Lipid 23
Lipid 24
wherein indicates the point of connection to the remainder of the compound.
26 . The delivery platform of claim 13 , wherein both X and Y are each independently selected from the group consisting of:
Name
Structure
Lipid 1
Lipid 2
Lipid 3
Lipid 4
Lipid 5
Lipid 6
Lipid 7
Lipid 8
Lipid 9
Lipid 10
Lipid 11
Lipid 12
Lipid 14
Lipid 15
Lipid 16
Lipid 17
Lipid 18
Lipid 19
Lipid 20
Lipid 21
Lipid 22
Lipid 23
Lipid 24
wherein indicates the point of connection to L 1 or L 2 .
27 . The delivery platform of claim 2 , wherein the PK/PD modulator is selected from the group consisting of:
or a pharmaceutically acceptable salt of any of these PK/PD modulators, wherein each indicates a point of connection to the RNAi agent.
28 . The delivery platform of claim 1 , wherein the RNAi agent inhibits expression of the mRNA of a human gene in a skeletal muscle cell.
29 . A composition comprising the delivery platform of claim 1 .
30 . A pharmaceutical composition comprising the composition of claim 29 and a pharmaceutical excipient.
31 . A method of treating a disease or disorder of a skeletal muscle cell comprising administering to a subject in need thereof a composition of claim 29 .
32 . The method of claim 31 , wherein the disease or disorder is muscular dystrophy.
33 . (canceled)