Preparation including vaccine adjuvant
Provided is a composition that is useful as a vaccine adjuvant and has excellent storage stability and immunostimulatory activity. Specifically provided is a freeze-dried preparation that has high storage stability, said preparation containing a (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, squalane, a hydrophilic surfactant, and an oleophilic surfactant, and being characterized by containing an ascorbic acid-based antioxidant and an excipient.
1 . A lyophilized formulation of an emulsion comprising:
i) (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 (Compound A), or a pharmaceutically acceptable salt thereof;
ii) squalane;
iii) an antioxidant A selected from the group consisting of an ascorbate ester, an inorganic salt of ascorbic acid, and ascorbic acid; and
iv) an excipient A selected from the group consisting of a non-reducing sugar and a sugar alcohol, provided that mannitol is excluded;
wherein the D 90 value of particle sizes of an emulsion reconstituted after 6-month storage at 5° C. as the lyophilized formulation is 1000 nm or less;
wherein the amount of squalane in the formulation is 200 to 300 times the weight of Compound A in the formulation;
wherein the amount of the antioxidant A in the formulation is 5 to 100 times the weight of Compound A in the formulation, in terms of the weight of sodium ascorbate; and
wherein the amount of the excipient A in the formulation is 200 to 625 times the weight of Compound A in the formulation.
2 . The formulation according to claim 1 , further comprising v) a hydrophilic surfactant and vi) a lipophilic surfactant.
3 . The formulation according to claim 1 , wherein the emulsion is an oil-in-water emulsion.
4 . The formulation according to claim 2 , wherein the hydrophilic surfactant is polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oils, or polyoxyethylene polyoxypropylene glycols.
5 . The formulation according to claim 2 , wherein the hydrophilic surfactant is Polysorbate 20, Polysorbate 40, Polysorbate 80, polyoxyethylene hydrogenated castor oil 60, or polyoxyethylene (160) polyoxypropylene (30) glycol.
6 . The formulation according to claim 2 , wherein the hydrophilic surfactant is Polysorbate 20, Polysorbate 40, or Polysorbate 80.
7 . The formulation according to claim 2 , wherein the lipophilic surfactant is sorbitan fatty acid esters, glycerin fatty acid esters sucrose fatty acid esters, or propylene glycol fatty acid esters.
8 . The formulation according to claim 2 , wherein the lipophilic surfactant is sorbitan fatty acid ester, sorbitan monooleate, sorbitan sesquioleate, or sorbitan trioleate.
9 . The formulation according to claim 2 , wherein the lipophilic surfactant is sorbitan trioleate.
10 . The formulation according to claim 1 , further comprising an antioxidant B selected from the group consisting of tocopherol, tocopherol acetate, and butyl hydroxyanisole.
11 . The formulation according to claim 10 , wherein the antioxidant B is α-tocopherol.
12 . The formulation according to claim 1 , wherein the antioxidant A is ascorbyl palmitate, potassium ascorbate, sodium ascorbate, or ascorbic acid.
13 . The formulation according to claim 1 , wherein the antioxidant A is sodium ascorbate or potassium ascorbate.
14 . The formulation according to claim 1 , wherein the excipient A is a non-reducing sugar selected from sucrose and trehalose or a sugar alcohol selected from sorbitol, erythritol, xylitol, maltitol, and lactitol.
15 . The formulation according to claim 2 , wherein the amount of the hydrophilic surfactant in the formulation is 0.5 to 250 times the weight of the (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, or a pharmaceutically acceptable salt thereof, in the formulation.
16 . The formulation according to claim 2 , wherein the amount of the lipophilic surfactant in the formulation is 0.5 to 250 times the weight of the (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, or a pharmaceutically acceptable salt thereof, in the formulation.
17 . The formulation according to claim 10 , wherein the amount of the blending antioxidant B in the formulation is 5 to 250 times the weight of the (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, or a pharmaceutically acceptable salt thereof, in the formulation.
18 . The formulation according to claim 1 , wherein the weight of the (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, or a pharmaceutically acceptable salt thereof, is 0.0001 to 0.65 times the weight obtained by excluding the (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, or a pharmaceutically acceptable salt thereof, from the lyophilized formulation.
19 . The formulation according to claim 1 , wherein both D 90 values of particle sizes of an emulsion immediately after preparation and an emulsion reconstituted after 6-month storage at 25° C. as the lyophilized formulation are 1000 nm or less.
20 . The formulation according to claim 1 , wherein an increased amount in the area percentage values of an impurity UK-1.02 after 6-month storage at 5° C. as the lyophilized formulation is 5.0% or less.
21 . A vaccine adjuvant comprising the formulation according to claim 1 .
22 . A vaccine comprising the formulation according to claim 1 and an antigen.
23 . The vaccine according to claim 22 , wherein the antigen is derived from a pathogen.
24 . A kit comprising the formulation according to claim 1 and an antigen.
25 . The formulation according to claim 1 , wherein a D 90 value of particle sizes of an emulsion reconstituted after 6-month storage at 25° C. as the lyophilized formulation is 2-fold or less than a D 90 value of particle sizes of an emulsion immediately after preparation.
26 . The formulation according to claim 1 , wherein a D 90 value of particle sizes of an emulsion during preparation is 1000 nm or less.
27 . The formulation according to claim 1 , wherein a D 90 value of particle sizes of an emulsion at the start time of lyophilization storage is 1000 nm or less.
28 . The formulation according to claim 1 , wherein a D 90 value of particle sizes of an emulsion reconstituted after 6-month storage at 25° C. as the lyophilized formulation is 1000 nm or less.