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 is C 1-4 alkyl;
R 2 is H or C 1-3 alkyl, wherein the C 1-3 alkyl is optionally substituted with hydroxy group;
R 3 is C 1-3 alkyl or C 1-3 alkoxy;
R 4 is H, halogen, hydroxy, a C 1-6 alkyl, or C 1-6 alkoxy;
Y 1 is a single bond and Y 2 is —C(O)—; or
Y 1 is —(CR 9 R 10 ) p — and Y 2 is a single bond;
L is a straight chain C 2-6 alkylene;
R 5 and R 6 are independently H or C 1-3 alkyl;
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. A kit comprising:
a) a liposome according to claim 1 ; and
b) an antigen.
11. The liposome according to claim 2 , 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 4 , 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.
13. The liposome according to claim 5 , 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.
14. The pharmaceutical composition according to claim 6 , 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.
15. The vaccine adjuvant according to claim 7 , 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.
16. The vaccine according to claim 8 , 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.
17. The vaccine according to claim 9 , 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.
18. The kit according to claim 10 , 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.