Polyanionic delivery of nucleic acids
Disclosed are dendritic anionic lipids which are compounds of Formula (I): wherein R and R 1 are non-polar groups, L is a linking moiety, and Dm is a dendritic moiety of m generations, each as defined herein. These dendritic anionic lipids are useful for delivery and expression of m RNA and encoded protein, e.g., as a component of liposomal delivery vehicle, and accordingly can be useful for treating various diseases, disorders and conditions, such as those associated with deficiency of one or more proteins.
1. A compound of formula I:
wherein:
each of R and R 1 independently comprise one or more non-polar groups selected from cholesterol, optionally substituted C 8 -C 26 aliphatic or optionally substituted C 8 -C 26 acyl;
L is —X 1 —, —X—R 2 —X— or —X—R 2 —R 2c —R 2 —X—, wherein:
each instance of R 2 independently is optionally substituted C 1 -C 10 aliphatic;
each instance of X or X 1 independently is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O—, —N(R 3 )—C(O)—N(R 3 )—, —SS—, or —S;
each instance of R 3 independently is —H or C 1-4 alkyl;
R 2c is a polyethylene glycol linker having a weight average molecular weight of from 350 Da to 45 kDa;
D m is a dendritic moiety of m generations, comprising
2 m -1 molecular units of formula
and
2 m molecular units of formula
m is an integer from 1 to 6;
in each generation independently, one of j and k is 1 and the other is 0 or 1; and
A is —OH or a pharmaceutically acceptable anion.
2. The compound of claim 1 , wherein:
A is —OSO 3 − .
3. The compound of claim 1 , wherein:
L is —X 1 —;
X 1 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—; and
each instance of R 3 independently is H or C 1-4 alkyl.
4. The compound of claim 3 , wherein:
X 1 is —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH—, —SS—, or —S.
5. The compound of claim 1 , wherein:
L is —X 2 —R 2a —X 3 —;
X 2 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—;
R 2a is C 1-10 aliphatic;
X 3 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—; and
each instance of R 3 is independently is —H or C 1-4 alkyl.
6. The compound of claim 1 , wherein:
L is —X 4 —R 2b —R 2c —R 2d —X 5 —;
X 4 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—;
R 2b is C 1-10 aliphatic;
R 2c is a polyethylene glycol linker having an average molecular weight of from 350 Da to 45 kDa
R 2d is C 1-10 aliphatic;
X 5 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—; and
each instance of R 3 independently is H or C 1-4 alkyl.
7. The compound of claim 6 , wherein:
X 4 is —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O— or —NH—C(O)—NH; and
X 5 is —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O— or —NH—C(O)—NH—.
8. The compound of claim 1 , wherein:
L is —X 4 —R 2b —R 2c —R 2d —X 5 —;
X 4 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—;
R 2b is C 1-10 aliphatic;
R 2c is —(OCH 2 CH 2 ) n —O—, wherein the average value of n is from 8 to 1000;
R 2d is C 1-10 aliphatic;
X 5 is —O—, —C(O)—, —S(O) 2 —, —C(O)—O—, —O—C(O)—, —C(O)—N(R 3 )—, —N(R 3 )—C(O)—, —O—C(O)—N(R 3 )—, —N(R 3 )—C(O)—O— or —N(R 3 )—C(O)—N(R 3 )—; and
each instance of R 3 is independently is H or C 1-4 alkyl.
9. The compound of claim 8 , wherein:
X 4 is —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O— or —NH—C(O)—NH—; and
X 5 is —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O— or —NH—C(O)—NH—.
10. The compound of claim 1 , wherein:
each instance of R 1 is
11. The compound of claim 1 , wherein m is an integer that is 1, 2, or 3.
12. The compound of claim 1 , wherein m is an integer that is 4, 5, or 6.
13. A composition comprising an mRNA encoding a protein, encapsulated within a liposome, wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids, wherein at least one non-cationic lipid is the compound of claim 1 .
14. The composition of claim 13 , comprising an mRNA encoding for cystic fibrosis transmembrane conductance regulator (CFTR) protein.
15. The composition of claim 13 , comprising an mRNA encoding for ornithine transcarbamylase (OTC) protein.
16. A composition comprising a nucleic acid encapsulated within a liposome, wherein the liposome comprises the compound of claim 1 .
17. The composition of claim 16 , further comprising one more lipids selected from the group consisting of one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids.
18. The composition of claim 16 , wherein the nucleic acid is an mRNA encoding a peptide or polypeptide.
19. The composition of claim 16 , wherein the mRNA encodes a peptide or polypeptide for use in the delivery to or treatment of the lung of a subject or a lung cell.
20. The composition of claim 19 , wherein the mRNA encodes cystic fibrosis transmembrane conductance regulator (CFTR) protein.