Methods of preparing a liquid formulation of G-CSF
Provided are pharmaceutical liquid formulations of G-CSF, which are stable over a long time period and substantially free of excipients, as well as ready-to-use syringes containing such formulations and corresponding kits.
1. A method of preparing a medicament comprising an aqueous liquid formulation of G-CSF, the aqueous liquid formulation comprising G-CSF, a sugar alcohol, a surfactant at a concentration of 0.05 to 0.06 mg/ml, acetate at a concentration of 5 to 20 mM as a buffer substance at a pH value of 4.15 to 4.3, and optionally amino acids and/or glycerol and/or carbohydrates and/or preservatives, wherein the method comprises mixing water, G-CSF, the sugar alcohol, the surfactant, and acetate in the form of its free acid to form a solution and adjusting the pH value of the solution by the addition of a base.
2. The method of claim 1 , wherein the acetate is provided in the form of acetic acid and the pH value of the solution is adjusted with sodium hydroxide solution.
3. The method according to claim 1 , wherein the G-CSF is non-glycosylated.
4. The method according to claim 1 , wherein the G-CSF is present at a concentration of 0.6 mg/ml.
5. The method according to claim 1 , wherein the surfactant is polysorbate.
6. The method according to claim 1 , wherein the sugar alcohol is sorbitol.
7. The method according to claim 1 , wherein the sugar alcohol is present at a concentration of 5% (w/v).
8. The method according to claim 1 , wherein the acetate is present at a concentration of about 10 mM.
9. The method according to claim 1 , wherein said medicament is substantially free of amino acids and/or additional protein.
10. The method according to claim 1 , wherein said medicament is a solution that is suitable for injection or infusion.
11. The method according to claim 10 , wherein said medicament comprises an aqueous liquid formulation of G-CSF, consisting of human, non-glycosylated methionyl G-CSF at a concentration of 0.6 mg/ml and of sorbitol at a concentration of 5% (w/v), polysorbate 80 at a concentration of 0.06 mg/ml, and acetate at a concentration of about 10 mM as a buffer substance at a pH value of 4.15 to 4.3.
12. The method of claim 1 , wherein said medicament further comprises an amino acid, glycerol, a carbohydrate, and/or a preservative and the method further comprises mixing the amino acid, glycerol, the carbohydrate, and/or the preservative with water, G-CSF, the sugar alcohol, the surfactant, and acetate.
13. The method of claim 11 , wherein said medicament contains less than 1% oxidized species after storage for 4 weeks at 40° C. or 5° C.