Compositions and methods for delivery of agents
This disclosure provides improved lipid-based compositions, including lipid nanoparticle compositions, and methods of use thereof for delivering agents in vivo including nucleic acids and proteins. These compositions are not subject to accelerated blood clearance and they have an improved toxicity profile in vivo.
1. A method for delivering a therapeutic level of a protein of interest to a subject, the method comprising:
administering to the subject at least two doses of lipid nanoparticles (LNPs) comprising an mRNA coding for the protein of interest, wherein the LNPs comprise an ionizable lipid, a helper lipid, a structural lipid, and a PEG-lipid,
wherein the PEG-lipid is a compound of Formula (V—OH):
or a pharmaceutically acceptable salt thereof, wherein:
r is an integer between 1 and 100, inclusive; and
R 5 is optionally substituted C 10-40 alkyl; and
wherein said LNPs result in a reduced anti-PEG IgM response in the subject as compared to LNPs lacking a PEG-lipid of Formula (V—OH).
2. The method of claim 1 , wherein the compound of Formula (V—OH) is Cmpd452:
3. The method of claim 2 , wherein the PEG-lipid comprises a PEG molecule of an average molecular weight of 2,000 Da or less.
4. The method of claim 3 , wherein the PEG-lipid is HO-PEG2000-ester-C18 (Cmpd403).
5. The method of claim 1 , wherein the ionizable lipid is a compound of Formula (XI):
or a pharmaceutically acceptable salt thereof, wherein:
l is selected from 1, 2, 3, 4, and 5;
M and M′ are independently selected from C(O)O, and OC(O);
M 1 is M′;
R 2 and R 3 are both C 1-14 alkyl, or C 2-14 alkenyl;
R 4 is (CH 2 ) n Q, in which Q is OH, and n is selected from 2, 3, or 4; and
R′ is a C 1 -C 18 linear alkyl.
6. The method of claim 5 , wherein R′ is C 9 alkyl.
7. The method of claim 5 , wherein n is 2.
8. The method of claim 5 , wherein the compound of Formula (XI) has the structure of Cmpd18:
9. The method of claim 1 , wherein the helper lipid comprises at least one fatty acid chain of at least 8 carbons in length and at least one polar headgroup moiety, and wherein the helper lipid does not comprise a phosphatidyl choline (PC) and is a zwitterionic non-cationic helper lipid, a DSPC analog, oleic acid, an oleic acid analog, DSPC, or a 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) substitute.
10. The method of claim 9 , wherein the helper lipid is DSPC.
11. The method of claim 1 , wherein the helper lipid is oleic acid or an oleic acid analog.
12. The method of claim 1 , wherein the LNPs comprise a molar ratio of about 45-65% ionizable lipid: 5-25% helper lipid: 15-45% structural lipid; and 0.15-15% PEG-lipid.
13. The method of claim 12 , wherein the PEG-lipid is present in the LNPs in an amount of less than 0.5% (w/w).
14. The method of claim 1 , wherein said administering results in a reduced IgG response in the subject.
15. The method of claim 1 , wherein said administering results in a reduced anti-DSPC IgM response in the subject.
16. The method of claim 1 , wherein said administering results in reduced B1a cell activation in the subject.
17. The method of claim 1 , wherein said administering results in reduced CD36 positive cell activation in the subject.
18. The method of claim 1 , wherein said at least two doses of LNPs is more than two doses of LNPs.
19. The method of claim 1 , wherein the interval between the doses is about 21 days or less.
20. The method of claim 1 , wherein the mRNA is chemically-modified.
21. The method of claim 1 , wherein the mRNA comprises at least one miR binding site.
22. The method of claim 21 , wherein the miR binding site is selected from miR 122, 126, miR 155, and miR 142.3p.
23. The method of claim 1 , further comprising administering to the subject an additional agent that inhibits immune responses.
24. The method of claim 1 , comprising:
administering to the subject a first dose of lipid nanoparticles (LNPs) comprising an mRNA coding for the protein of interest, wherein the LNPs comprise an ionizable lipid, a helper lipid, a structural lipid, and a PEG-lipid, wherein:
the PEG-lipid
is Cmpd452:
the ionizable lipid is a compound of Formula (XI):
or a pharmaceutically acceptable salt thereof, wherein:
l is selected from 1, 2, 3, 4, and 5;
M and M′ are independently selected from C(O)O, and OC(O);
M 1 is M′;
R 2 and R 3 are both C 1-14 alkyl, or C 2-14 alkenyl;
R 4 is (CH 2 ) n Q, in which Q is OH, and n is selected from 2, 3, or 4; and
R′ is a C 1 -C 18 linear alkyl;
the helper lipid comprises at least one fatty acid chain of at least 8 carbons in length and at least one polar headgroup moiety, and wherein the helper lipid does not comprise a phosphatidyl choline (PC) and is a zwitterionic non-cationic helper lipid, a DSPC analog, oleic acid, an oleic acid analog, DSPC, or a 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) substitute; and
the structural lipid is a steroid; and
administering a second dose of said LNPs;
wherein said LNPs result in a reduced anti-PEG IgM response in the subject as compared to LNPs lacking a PEG-lipid of Formula (V—OH).
25. The method of claim 24 , wherein the PEG-lipid is HO-PEG2000-ester-C18(Cmpd403).
26. The method of claim 24 , wherein the compound of Formula (XI) has the structure of Cmpd18:
27. The method of claim 24 , wherein the helper lipid is DSPC.
28. The method of claim 24 , wherein the structural lipid is cholesterol.