Formulations of PEG-functionalised serine proteases with high concentrations of an aromatic preservative
An aromatic preservative and functionalization technology, applied in medical preparations containing active ingredients, medical preparations with inactive ingredients, peptide/protein ingredients, etc., can solve the problem of reducing the solubility of factor VII polypeptides
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment approach
[0113] The inventors have now identified particularly advantageous embodiments, namely a liquid, aqueous pharmaceutical composition as defined herein, comprising:
[0114] (i) 1-90 mg / mL of a Factor VII polypeptide functionalized with one or more polyethylene glycol (PEG) moieties having a molecular weight of 500-60,000 Da;
[0115] (ii) a buffer suitable for maintaining a pH in the range of about 5.0 to about 9.0;
[0116] (iii) at least one aromatic preservative in a concentration of at least 0.1-20 mg / mL; and
[0117] (iv) at least one antioxidant at a concentration of at least 0.1-5.0 mg / mL.
[0118] In another preferred embodiment, said liquid, aqueous pharmaceutical composition comprises:
[0119] (i) 40K-PEG-rFVIIa,
[0120] (ii) a buffer to maintain the pH in the range of about 5 to about 6, and
[0121] (iii) Phenol at a concentration of 1.0-10.0 mg / mL or m-cresol at a concentration of 1.0-5.0 mg / mL.
[0122] In a third preferred embodiment, said liquid, aqueous ...
Embodiment 1
[0191] A solution of rFVIIa and 10K-PEG-rFVIIa was buffer exchanged on a NAP10 column to obtain a solution containing 6.25 mM CaCl at pH 6.0 2 and 6.25 mM histidine buffer. Then, a sample containing 20 μl of rFVIIa or 10K-PEG-rFVIIa solution and 5 μl of 5 mg / mL m-cresol, 10 mg / mL m-cresol, 15 mg / mL m-cresol or 30 mg / mL phenol was prepared. The final concentration is approximately 2.6 mg / mL rFVIIa or 2.6 mg / mL 10K-PEG-rFVIIa, 5 mM CaCl 2 , 5 mM histidine and 1, 2 or 3 mg / mL m-cresol or 6 mg / mL phenol. Samples were briefly shaken (vortexed) and transferred to a 15 μl cuvette with a 1.5 mm pathlength. Then, turbidity at 400 nm was measured as absorbance. High turbidity is indicative of a precipitate of large aggregates of FVII polypeptide. From this test, it can be seen that in the presence of preservatives, solutions of rFVIIa showed significant turbidity, indicating that m-cresol and phenol induced precipitation of the sample (see figure 1 ). On the other hand, in the pre...
Embodiment 2
[0194] With 10mM His, pH 5.5, 20mM CaCl 2 , 6% sucrose and 0.5 mg / mL methionine to prepare 22 mg / mL of 40K-PEG-rFVIIa. Samples of 200 μl were lyophilized and reconstituted in 3 mg / mL m-cresol or 15 mg / mL benzyl alcohol. Immediately after reconstitution, a 20 μl sample was withdrawn and incubated in 10 mM His, pH 5.5, 20 mM CaCl 2 , Diluted to 1mg / mL in 6% sucrose. These reference samples with low preservative content were then stored at 4°C. A sample containing 3 mg / mL m-cresol and 15 mg / mL benzyl alcohol was also stored at 4°C. After 5 weeks, another 20 μl was withdrawn and diluted to 1 mg / mL. These samples and reference samples were then assayed by size exclusion chromatography, clot activity and FX activation. Table 1 shows the analytical results, coagulation activity and FX activation are given relative to the values obtained for the reference samples. These results show that the presence of high concentrations of preservatives caused little sample degradation.
...
PUM
| Property | Measurement | Unit |
|---|---|---|
| molecular weight | aaaaa | aaaaa |
| molecular weight | aaaaa | aaaaa |
| concentration | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More