Vaccine adjuvant formulation
The present invention provides a liposome useful as a vaccine adjuvant. More specifically, the present invention provides a liposome which comprises a lipid bilayer comprising dimyristoylphosphatidylcholine and egg phosphatidylglycerol and a conjugated compound in which a low-molecular weight compound enhancing the physiological activity of TLR7 is bound to squalene via a linker, said conjugated compound being encapsulated in the lipid bilayer.
1. A liposome comprising (1) and (2):
(1) a lipid multilayer comprising a mixture of two different lipids selected from the group consisting of: hydrogenated soybean phosphatidylcholine (HSPC), dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylglycerol (DOPG), dimyristoylphosphatidylcholine (DMPC) and egg yolk phosphatidylglycerol (EPG), or a mixture of the two different lipids and cholesterol (Chol); and
(2) a conjugated compound encapsulated in the lipid multilayer of (1), wherein the conjugated compound is a compound of formula (1):
or a pharmaceutically acceptable salt thereof, wherein
X is —CH 2 —;
R 1 and R 2 are independently H or C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with 1, 2, 3 or 4 substituents independently selected from the group consisting of hydroxy, halogen and C 1-6 alkoxy;
R 3 is C 1-6 alkyl, C 1-6 alkoxy or C 1-6 alkylthio;
R 4 is H, halogen, hydroxy, C 1-6 alkyl, C 1-6 alkoxy or cyano;
Y 1 is a single bond or —(CR 9 R 10 ) p —;
Y 2 is a single bond or —C(O)—;
L is a straight chain C 2-6 alkylene, wherein the C 2-6 alkylene is optionally substituted with 1, 2, 3 or 4 substituents independently selected from the group consisting of C 1-5 alkyl, hydroxy, and halogen;
R 5 and R 6 are independently H or C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with 1, 2, 3 or 4 substituents independently selected from the group consisting of hydroxy and halogen;
or R 5 and R 6 are taken together with -N-L-N-, to form piperazin-1,4-diyl, wherein the piperazin-1,4-diyl is optionally substituted with 1, 2, 3 or 4 substituents independently selected from the group consisting of methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, oxo, hydroxy and halogen;
R 9 and R 10 are independently selected from H and C 1-4 alkyl;
m is 1; and
p is 1.
2. The liposome according to claim 1 , wherein the weight ratio of the conjugated compound to the lipid components is 1:8 to 1:250.
3. The liposome comprising a conjugated compound according to claim 1 , wherein the lipid multilayer of (1) is a lipid multilayer comprising lipid components comprising a mixture of dimyristoylphosphatidylcholine (DMPC) and egg yolk phosphatidylglycerol (EPG).
4. The liposome according to claim 1 , wherein the molar ratio of dimyristoylphosphatidylcholine (DMPC) to egg yolk phosphatidylglycerol (EPG) is 1:1 to 2:1.
5. The liposome according to claim 1 , wherein the lipid multilayer is a lipid bilayer.
6. A pharmaceutical composition comprising the liposome according to claim 1 .
7. A vaccine adjuvant comprising the liposome according to claim 1 .
8. A vaccine comprising the liposome according to claim 1 , and an antigen.
9. The vaccine according to claim 8 , wherein the antigen is a substance derived from a pathogen.
10. The liposome according to claim 1 , wherein the conjugated compound is
(4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenamide;
(4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3- methoxybenzyl}piperazin-1-yl)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaen-1-one;
(4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3- methoxybenzoyl}piperazin-1-yl)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaen-hexamethylhexacosa-4,8,12,16,20,24-hexaen-1-one;
4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenoyl]amino}ethyl)-3-methoxybenzamide; or
4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenoyl](methyl)amino}ethyl)-3-methoxybenzamide.
11. The liposome according to claim 1 , wherein the conjugated compound is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenamide.
12. The liposome according to claim 1 , wherein:
the lipid multilayer of (1) comprises a mixture of dimyristoylphosphatidylcholine (DMPC) and egg yolk phosphatidylglycerol (EPG); and
the conjugated compound of (2) being encapsulated in the lipid multilayer of (1) is
(4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenamide;
(4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3- methoxybenzyl}piperazin-1-yl)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaen-1-one;
(4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3- methoxybenzoyl}piperazin-1-yl)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaen-hexamethylhexacosa-4,8,12,16,20,24-hexaen-1-one;
4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenoyl]amino}ethyl)-3-methoxybenzamide; or
4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenoyl](methyl)amino}ethyl)-3-methoxybenzamide.
13. The liposome according to claim 1 , wherein:
the lipid multilayer of (1) comprises a mixture of dimyristoylphosphatidylcholine (DMPC) and egg yolk phosphatidylglycerol (EPG); and
the conjugated compound of (2) being encapsulated in the lipid multilayer of (1) is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacosa-4,8,12,16,20,24-hexaenamide.
14. A kit comprising:
a) a liposome according to claim 1 ; and
b) an antigen.
15. The liposome according to claim 1 , wherein:
Y 1 is a —(CR 9 R 10 ) p — and p is 1;
Y 2 is a single bond;
R 1 is C 1-6 alkyl;
R 2 is C 1-6 alkyl substituted with 1, 2, 3, or 4 hydroxy;
R 3 is C 1-6 alkyl;
R 4 , R 6 , R 9 , and R 10 are each hydrogen;
R 5 is C 1-6 alkyl;
L is a straight chain C 2-6 alkylene;
m is 1; and
p is 1.