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 said mRNA comprises an open reading frame encoding a cystic fibrosis transmembrane regulator (CFTR) protein, wherein said lipid composition comprises:
i. a zwitterionic selective organ targeting (SORT) lipid at a molar percentage from about 5% to about 50%;
ii. an ionizable cationic lipid at a molar percentage of at least about 5%, wherein the ionizable cationic lipid comprises an ammonium group that is positively charged at a pH from about 6 to about 8 and at least two C 6 -C 24 alkyl or alkenyl groups;
iii. a steroid or steroid derivative at a molar percentage from about 15% to about 46%; and
iv. a polymer-conjugated lipid at a molar percentage from about 0.5% to about 10%;
wherein said molar percentage is determined based on the total lipids present in said lipid composition; wherein said composition is formulated for inhalation for delivery to a lung.
2. The composition of claim 1 , wherein:
i. said zwitterionic SORT lipid is distearoylphosphatidylcholine (DSPC) or 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl ethyl phosphate (DOCPe) and is present at a molar percentage from about 10% to about 45%;
ii. said ionizable cationic lipid is present at a molar percentage of at least about 20%;
iii. said steroid or steroid derivative is cholesterol and is present at a molar percentage from about 20% to about 46%; and
iv. said polymer-conjugated lipid is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) and is present at a molar percentage from about 0.5% to about 4%.
3. The composition of claim 1 , wherein said lipid composition is characterized by an apparent ionization constant (pKa) outside a range of about 6 to about 7 as determined by a 2-(p-toluidino)-6-naphthalenesulfonic acid titration assay.
4. The composition of claim 3 , wherein said pKa of said lipid composition is about 8 or higher, or of about 6 or lower.
5. 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 zwitterionic SORT lipid.
6. The composition of claim 1 , wherein said composition is configured to deliver a greater amount said mRNA in said lung as compared to that achieved absent said zwitterionic SORT lipid.
7. The composition of claim 1 , wherein when said composition is administered to a subject, a surface of said lipid composition interacts with an apolipoprotein E (Apo E) to a lesser degree than with an endogenous protein that is not Apo E.
8. The composition of claim 1 , wherein said composition further comprises an endogenous protein.
9. The composition of claim 1 , wherein said composition further comprises a non-endogenous protein.
10. The composition of claim 1 , wherein said composition further comprises a fluorescent protein.
11. The composition of claim 10 , wherein said fluorescent protein is a td-Tomato.
12. The composition of claim 1 , wherein said lipid composition further comprises a permanently cationic SORT lipid.
13. The composition of claim 12 , wherein said permanently cationic SORT lipid comprises a quaternary ammonium ion.
14. The composition of claim 12 , wherein said permanently cationic SORT lipid has 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 (C1-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.
15. The composition of claim 12 , wherein said permanently cationic SORT lipid is further defined as:
16. The composition of claim 12 , wherein said permanently cationic SORT lipid comprises 1,2-Dioleoyl-3-trimethylammonium propane (DOTAP).
17. The composition of claim 1 , wherein said ionizable cationic lipid is present at a molar percentage of about 30%.
18. The composition of claim 1 , wherein said ionizable cationic lipid is present at a molar percentage of about 5% to about 30%.
19. The composition of claim 1 , wherein:
(a) Said zwitterionic SORT lipid is distearoylphosphatidylcholine (DSPC) or 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl ethyl phosphate (DOCPe), and wherein said zwitterionic SORT lipid is present at a molar percentage from about 10% to about 40%;
(b) said ionizable cationic lipid is present at a molar percentage of at least about 20%;
(c) said steroid or steroid derivative is a cholesterol, and wherein said cholesterol is present at a molar percentage from about 20% to about 46%; and
(d) said polymer-conjugated lipid is a 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) and is present at a molar percentage from about 0.5% to about 4%,
wherein said molar percentage is determined based on the total lipids present in said lipid composition; wherein said composition is formulated for inhalation for delivery a lung.
20. The composition of claim 1 , wherein:
(a) said zwitterionic SORT lipid is distearoylphosphatidylcholine (DSPC) or 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl ethyl phosphate (DOCPe), and wherein said zwitterionic SORT lipid is present at a molar percentage from about 10% to about 40%;
(b) said ionizable cationic lipid is present at a molar percentage of at least about 20%;
(c) said steroid or steroid derivative is present at a molar percentage from about 20% to about 46%; wherein the steroid or steroid derivative comprises a ring structure as shown
and is optionally substituted with alkyl groups, alkoxy groups, hydroxy groups, oxo groups, acyl groups, or a double bond between two or more carbon atoms;
(d) said polymer-conjugated lipid is a 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) and is present at a molar percentage from about 0.5% to about 4%, and
wherein said molar percentage is determined based on the total lipids present in said lipid composition; wherein said composition is formulated for inhalation for delivery to a lung.
21. The composition of claim 1 , wherein:
(a) said zwitterionic SORT lipid is distearoylphosphatidylcholine (DSPC) or 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl ethyl phosphate (DOCPe), wherein said zwitterionic SORT lipid is present at a molar percentage from about 10% to about 40%;
(b) said ionizable cationic lipid is present at a molar percentage of at least about 20%;
(c) said steroid or steroid derivative is present at a molar percentage from about 20% to about 46%; and
(d) said polymer conjugated lipid is represented by Formula (B) and is present at a molar percentage from about 0.5% to about 4%:
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,
wherein said molar percentage is determined based on the total lipids present in said lipid composition; wherein said composition is formulated for inhalation for delivery to a lung.