Ionizable lipids for nucleic acid delivery
The present document describes compounds, or pharmaceutically acceptable salt thereof, of a core formula (I) where R 1 features an amine group, particularly useful in the formulation of lipid particles including nucleic acid therapeutic agents, or proteins, or both, and for delivery of nucleic acid and protein therapeutics to cells in vivo or ex vivo, including anticancer and vaccine applications.
1 . A compound, or a pharmaceutically acceptable salt thereof, of formula (I):
wherein:
L 1 is a direct bond or C 1 -C 5 alkylene;
E 1 is —O—, —OC(O)O—, —OC(O)-δ 1 , —OC(O)N(Q)-δ 1 , —OC(O)S-δ 1 , —N(Q)C(O)-δ 1 , —N(Q)C(O)O-δ 1 , —C(O)O-δ 1 , or —C(O)N(Q)-δ 1 ; Q is H or C 1 -C 5 alkyl; δ 1 designates the bond linked to the R 1 group;
R 1 is selected from:
and wherein:
R 4 and R 5 are each independently C 1 -C 6 alkyl, C 2 -C 6 alkenyl, or C 2 -C 6 alkynyl; alternatively R 4 and R 5 may join to form 4-6 membered heterocyclic ring containing oxygen (O) or up to 2 nitrogen (N),
optionally substituted with 1 or 2 substituents that are each independently C 1 -C 6 alkyl, cyclopropyl, OH, and C 1 -C 3 alkoxy;
R 6 is C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 3 -C 6 cycloalkyl or 2-hydroxyethyl;
R 7 is H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, or C 2 -C 6 alkynyl;
a and c′ are each independently 1, 2, 3, 4, or 5;
b, c and e are each independently 0, 1, or 2;
d is 1, or 2;
R 2 is H, C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 2 -C 12 alkynyl, or
L 2 is a direct bond or δ 2 —(CR 8 R 8′ ) k -δ 3 wherein R 8 and R 8′ are each independently H, C 1 -C 12 alkyl, C 2 -C 12 alkenyl or C 2 -C 12 alkynyl; δ 2 designates the bond linked to E 2 group, and δ 3 designates the bond linked to cyclopentyl scaffold described in formula (I);
k is 1, 2, 3, 4, or 5;
E 2 is —O—, —OC(O)O—, —OC(O)-δ 4 , —OC(O)N(Q)-δ 4 , —N(Q)C(O)-δ 4 , —N(Q)C(O)O-δ 4 , —C(O)N(Q)-δ 4 or —C(O)O-δ 4 ; Q is H or C 1 -C 5 alkyl; where δ 4 designates the bond linked to R 3 group;
R 3 is C 8 -C 20 alkyl, C 8 -C 20 alkenyl, C 8 -C 20 alkynyl,
wherein:
f is 0 or 1;
g is 1 or 2;
g′ is 1, 2, 3, 4, or 5;
h is 0, 1, 2, 3 or 4;
R 9 is a C 6 -C 20 chain having the formula —(CH 2 ) i -[L 4 -(CH 2 )] j —R 12 , wherein:
L 4 is selected from
i is an integer in the range 6-20;
j is 0, 1, 2, or 3;
R 12 is H or C 4 -C 8 alkyl;
R 9′ is H, C 4 -C 10 alkyl, C 4 -C 10 alkenyl, or C 4 -C 10 alkynyl;
R 10 and R 10′ are each independently C 4 -C 10 alkyl, C 4 -C 10 alkenyl, or C 4 -C 10 alkynyl;
L 3 is —OC(O)—δ 5 , —O-δ 5 , or a direct bond;
δ 5 designates the bond linked to R 10 and R 10′ ; and
R 11 =R 9 , or has the formula:
2 . A compound or a pharmaceutically acceptable salt thereof, of formula (I):
wherein:
L 1 is a direct bond or C 1 -C 5 alkylene;
E 1 is —OC(O)O—, —OC(O)-δ 1 , —OC(O)N(Q)-δ 1 or —OC(O)S-δ 1 ; Q is H or C 1 -C 5 alkyl; δ 1 designates the bond linked to the R 1 group;
R 1 is selected from:
wherein:
R 4 and R 5 are each independently C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl; alternatively R 4 and R 5 may join to form 5-6 membered heterocyclic ring containing up to 2 nitrogen (N), optionally substituted with 1 or 2 substituents that are each independently C 1 -C 6 alkyl, or cyclopropyl;
R 6 is C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl;
R 7 is H or C 1 -C 6 alkyl;
a is 1, 2, or 3;
b and c are each independently 0, 1, or 2;
c′ is 2, 3, or 4;
d is 1 or 2;
e is 0 or 1;
R 2 is H, C 1 -C 5 alkyl, C 2 -C 5 alkenyl, C 2 -C 5 alkynyl, or
L 2 is a direct bond or δ 2 —(CR 8 R 8′ ) k -δ 3 wherein R 8 and R 8′ are each independently H, C 1 -C 12 alkyl, C 2 -C 12 alkenyl or C 2 -C 12 alkynyl;
δ 2 designates the bond linked to E 2 group and δ 3 designates the bond linked to cyclopentyl scaffold described in formula (I);
k is 1;
E 2 is selected from —O—, —OC(O)O—, —OC(O)-δ 4 , —OC(O)N(Q)-δ 4 , —C(O)N(Q)-δ 4 or —C(O)O-δ 4 ; Q is H or C 1 -C 5 alkyl; where δ 4 designates the bond linked to R 3 group;
R 3 is selected from C 8 -C 20 alkyl, C 8 -C 20 alkenyl, C 8 -C 20 alkynyl,
wherein:
f and h are each 0;
g is 1 or 2;
h is 0, 1, 2, 3 or 4;
R 9 is a C 6 -C 20 chain having the formula —(CH 2 ) i -[L 4 -(CH 2 )] j —R 12 , wherein:
L 4 is selected from
i is an integer from 6-20;
j is 0, 1, or 2;
R 12 is H or C 4 -C 8 alkyl group;
R 9′ is H or C 4 -C 10 alkyl;
R 10 and R 10′ are each independently C 4 -C 10 alkyl, C 4 -C 10 alkenyl, or C 4 -C 10 alkynyl;
L 3 is —OC(O)-δ 5 or a direct bond; δ 5 designates the bond linked to R 10 and R 10′ ; and
R 11 is the same as R 9 .
3 . The compound of claim 1 , or a pharmaceutically acceptable salt thereof, of formula (I),
wherein:
L 1 is a direct bond;
E 1 is —OC(O)O—, —OC(O)-δ 1 , —OC(O)N(Q)-δ 1 , or —OC(O)S-δ 1 ; Q is H or C 1 -C 5 alkyl; δ 1 designates the bond linked to the R 1 group;
R 1 is selected from
wherein:
R 4 and R 5 are each, independently C 1 -C 6 alkyl; alternatively R 4 and R 5 may join to form 5-6 membered heterocyclic ring containing up to 2 nitrogen (N), optionally substituted with 1 or 2 substituents that are each independently a C 1 -C 6 alkyl;
R 6 is C 1 -C 6 alkyl or cyclopropyl;
R 7 is H or C 1 -C 6 alkyl;
a is 1, 2, or 3;
b is 0 or 1;
c is 0, 1, or 2;
c′ is 2, 3, or 4;
d is 2;
e is 1;
R 2 is H, C 1 -C 5 alkyl, C 2 -C 5 alkenyl,
L 2 is δ 2 —(CH 2 ) k -δ 3 , wherein δ 2 designates the bond linked to E 2 group, and δ 3 designates the bond linked to cyclopentyl scaffold described in formula (I);
k is 1;
E 2 is —C(O)O-δ 4 , where δ 4 designates the bond linked to R 3 group;
R 3 is selected from:
wherein:
f and h are each 0;
g is 1 or 2;
R 9 is a C 6 -C 20 chain having the formula —(CH 2 ) i -[L 4 -(CH 2 )] j —R 12 , wherein:
L 4 is selected from
i is an integer in the range 6-20;
j is 0, 1, or 2;
R 12 is H or C 4 -C 8 alkyl group;
R 9′ is H or C 4 -C 10 alkyl;
R 10 and R 10′ are each independently C 4 -C 10 alkyl;
L 3 is a direct bond;
R 11 is the same as R 9 .
4 . The compound of claim 1 , or a pharmaceutically acceptable salt thereof, of formula (I),
wherein:
L 1 is a direct bond;
E 1 is —OC(O)-δ 1 , wherein δ 1 designates the bond linked to the R 1 group;
R 1 is selected from:
wherein:
R 4 and R 5 are each independently C 1 -C 6 alkyl; alternatively, R 4 and R 5 may join to form a 5-6 membered heterocyclic ring containing up to 2 nitrogen (N), optionally substituted with 1 or 2 substituents that are each independently C 1 -C 6 alkyl;
R 6 is a C 1 -C 6 alkyl or cyclopropyl;
R 7 is H or C 1 -C 6 alkyl;
a is 1, 2, or 3;
b is 0, or 1;
c is 0, 1, or 2;
c′ is 2, 3, or 4;
d is 2;
e is 1;
R 2 is selected from H, C 1 -C 5 alkyl, C 2 -C 5 alkenyl, and
L 2 is δ 2 —(CH 2 ) k -δ 3 ; δ 2 designates the bond linked to E 2 group, and δ 3 designates the bond linked to cyclopentyl scaffold described in formula (I);
k is 1;
E 2 is —C(O)O-δ 4 , where δ 4 designates the bond linked to R 3 group;
R 3 is selected from:
wherein:
f and h are 0;
g is 1 or 2;
R 9 is a C 6 -C 20 chain having the formula —(CH 2 ) i [L 4 -(CH 2 )] j —R 12 , wherein:
L 4 is selected from
i is an integer in the range 6-20;
j is 0, 1, or 2;
R 12 is H or C 4 -C 8 alkyl;
R 9′ is H or C 4 -C 10 alkyl;
R 10 and R 10′ are each independently C 4 -C 10 alkyl;
L 3 is a direct bond;
R 11 is the same as R 9 .
5 . The compound of claim 1 , wherein the compound is selected from the group consisting of:
or a pharmaceutically acceptable salt thereof.
6 . The compound of claim 1 , wherein the compound is selected from the group consisting of:
7 . A lipid mix composition comprising the compound of claim 1 and a structural lipid.
8 . The lipid mix composition of claim 7 , wherein the structural lipid comprises DSPC, DSPE, DPPC, DMPC, DOPC, POPC, DOPE, SM, or a combination thereof.
9 . The lipid mix composition of claim 7 , further comprising a stabilizing agent.
10 . The lipid mix composition of claim 9 , further comprising a sterol, wherein the molar ratio of the compound to the rest of the components in the lipid mix composition is 30 Mol % to 70 Mol %.
11 . The lipid mix composition of claim 7 , wherein the experimental pKa of the lipid mix composition is in the range 5.6-7.1.
12 . A pharmaceutical composition comprising a compound of claim 1 and at least one pharmaceutically acceptable carrier or excipient.
13 . The lipid mix composition of claim 9 , further comprising a sterol, wherein the compound is present at about 40 Mol %-49 Mol %, the structural lipid is present at about 11-20 Mol %, the sterol is present at about 37.5 Mol %, and the stabilizing agent is present at about 2.5 Mol %.
14 . The lipid mix composition of claim 9 , wherein the stabilizing agent includes PEG-DMG 2000, Polyoxyethylene (10) stearyl ether, polyoxyethylene (40) stearate, Polysorbate 80, Polyoxyethylene (4) lauryl ether, Polyoxyethylene (20) stearyl ether, Polyoxyethylene (23) lauryl ether, or D-α-Tocopherol polyethylene glycol 1000 succinate.
15 . The compound of claim 5 , wherein one of the hydrogens is substituted with a halogen.
16 . The composition of claim 9 , wherein the stabilizing agent comprises a surfactant and a polymer conjugated lipid.