COMPOSITIONS AND METHODS FOR ORGAN SPECIFIC DELIVERY OF NUCLEIC ACIDS
The present disclosure provides compositions which shown preferential targeting or delivery of a nucleic acid composition to a particular organ. In some embodiments, the composition comprises a steroid or sterol, an ionizable cationic lipid, a phospholipid, a PEG lipid, and a permanently cationic lipid which may be used to deliver a nucleic acid.
1 . A composition comprising a messenger ribonucleic acid (mRNA) with a lipid composition, wherein the mRNA comprises an open reading frame encoding a cystic fibrosis transmembrane regulator (CFTR) protein, wherein said lipid composition comprises:
(a) a zwitterionic selective organ targeting (SORT) lipid at a molar percentage from about 5% to about 50%;
(b) an ionizable cationic lipid at a molar percentage of at least about 5%, wherein said ionizable cationic lipid comprises at least two alkyl (C6-C24) or alkenyl (C6-C24) groups, wherein when the pH of a solution comprising said ionizable cationic lipid is from about 6 to about 8, said ionizable cationic lipid comprises an ammonium group that is positively charged; and
wherein said molar percentage is determined based on the total lipids present in said lipid composition.
2 . The composition of claim 1 , wherein said composition is formulated for inhalation administration for delivery of said composition to a lung.
3 . The composition of claim 1 , wherein said ionizable cationic lipid comprises at least two alkenyl (C6-C24) groups.
4 . The composition of claim 1 , wherein said ionizable cationic lipid is present at a molar percentage of at least about 20%.
5 . The composition of claim 1 , wherein said zwitterionic SORT lipid is distearoylphosphatidylcholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), or 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl ethyl phosphate (DOCPe).
6 . The composition of claim 1 , wherein said composition further comprises a polymer-conjugated lipid.
7 . The composition of claim 6 , wherein said polymer-conjugated lipid is represented by Formula (B):
wherein
R 12 and R 13 are each independently alkyl (C≤24) , alkenyl (C≤24) , or a substituted version of either of these groups;
R e is hydrogen, alkyl (C≤8) , or substituted alkyl (C≤8) ; and
x is 1 to 250.
8 . The composition of claim 6 , wherein said polymer-conjugated lipid is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000).
9 . The composition of claim 1 , wherein said lipid composition is characterized by an apparent ionization constant outside a range of about 6 to about 7 pKa as determined by a 2-(p-toluidino)-6-naphthalenesulfonic acid titration assay.
10 . The composition of claim 1 , wherein said composition is configured to provide a greater amount of said mRNA in said lung as compared to that achieved absent said SORT lipid.
11 . The composition of claim 1 , wherein when said composition is administered to a subject, a surface of said lipid composition interacts with apolipoprotein E (Apo E) to a lesser degree than with an endogenous protein that is not Apo E.
12 . The composition of claim 1 , wherein said composition further comprises an endogenous protein.
13 . The composition of claim 1 , wherein said composition further comprises a non-endogenous protein.
14 . The composition of claim 1 , wherein said composition further comprises a fluorescent protein.
15 . The composition of claim 14 , wherein said fluorescent protein is td-Tomato.
16 . The composition of claim 1 , wherein said ionizable cationic lipid is present at a molar percentage of about 5% to about 30%.
17 . The composition of claim 1 , wherein said lipid composition further comprises a permanently cationic SORT lipid.
18 . The composition of claim 17 , wherein said permanently cationic SORT lipid comprise a quaternary ammonium cation.
19 . The composition of claim 18 , wherein said permanently cationic SORT lipid is represented by a structure of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, stereoisomer, tautomer thereof:
wherein, in Formula (I):
R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;
R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ; and
X − is a monovalent anion;
wherein, in Formula (II):
R 4 and R 4 ′ are each independently alkyl (C6-C24) , alkenyl (C6-C24) , or a substituted version of either group;
R 4 ″ is alkyl (C≤24) , alkenyl (c≤24) , or a substituted version of either group;
R 4 ′″ is alkyl (C2-C8) , alkenyl (C2-C8) , or a substituted version of either group; and
X 2 − is a monovalent anion; and
wherein, in Formula (III):
R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;
R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ;
R 4 is alkyl (C≤6) or substituted alkyl (C≤6) ; and
X − is a monovalent anion.
20 . The composition of claim 19 , wherein said permanently cationic lipid is further defined as: