Liquid composition of modified factor vii polypeptides
A composition and factor technology, which is applied in the direction of non-active ingredients of polymer compounds, drug combinations, blood coagulation/fibrinolysis factors, etc., can solve the problems of pain and discomfort of patients
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
preparation example Construction
[0105] Preparation and Purification of Modified Factor VII Polypeptides:
[0106] Modified Factor VII molecules suitable for formulation according to the invention and their production have been described in WO 92 / 15686, WO 94 / 27631 , WO 96 / 12800 and WO 97 / 47651.
[0107] In general, purified human Factor VIIa is preferably prepared by recombinant DNA techniques, as described by Hagen et al., Proc. ) mentioned.
[0108] Factor VII can also be produced using the methods described by Broze and Majerus, J. Biol. Chem. 255(4):1242-1247, 1980, and Hedner and Kisiel, J. Clin.invest. Factor VII produced by these methods does not contain detectable amounts of other coagulation factors. A further purified Factor VII preparation can be obtained with an additional gel filtration step as a final purification step. Factor VII is then converted to activated Factor Vila using known methods, for example using several different plasma proteins, such as Factor Xlla, IXa or Xa. In addition, ...
Embodiment 1
[0117] A. Determination method
[0118] Aggregate content was determined by non-denaturing size exclusion HPLC. The content of the oxidized form was determined by RP-HPLC. The content of enzymatically degraded forms was determined by RP-HPLC.
[0119] Native size exclusion chromatography was performed on a 7.5×300 mm Waters Protein Pak 300SW column using 0.2M ammonium sulfate, 5% 2-propanol pH 7.0 as the mobile phase. Flow rate: 0.5ml / min. Detection: 215nm. Loading: 25 μg FVIIa.
[0120] Reverse phase HPLC was performed on a proprietary 4.5 x 250 mm butyl bonded silica column with a particle size of 5 μm and a pore size of 300 Å. Column temperature: 70°C. A-buffer: 0.1% v / v trifluoroacetic acid. B-buffer: 0.09% v / v trifluoroacetic acid, 80% v / v acetonitrile. The column was eluted with a linear gradient from X to (X+13)% B in 30 minutes. X was adjusted so that FVIIa elutes at a retention time of approximately 26 minutes. Flow rate: 1.0ml / min. Detection: 214nm. Loadi...
Embodiment 2
[0122] Chemical stability of aqueous formulations of Phe-Phe-Arg chloromethyl ketone-inactivated factor VII (FFR-rFVIIa) containing methionine as an antioxidant
[0123] Two different formulations were prepared. The composition of the preparation is as follows:
[0124] FFR-rFVIIa 2mg / ml
[0125] NaCl 2.8-2.9mg / ml
[0126] CaCl2 , 2H 2 O 1.4-1.5mg / ml
[0127] Glycylglycine 1.3mg / ml
[0128] Methionine 0 or 1mg / ml
[0129] pH 7.0
[0130] Methionine (mg / ml)
[0131] The results showed that the addition of methionine slowed down the oxidation rate of the preparation.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More