Formulation Comprising Bioactive Agents And Method Of Using Same
a technology of bioactive agents and forms, which is applied in the field of compositions containing bioactive agents, can solve the problems of affecting the development of pharmaceutical compositions containing this protein, and affecting the development of pharmaceutical compositions
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example 1
Preparation of BMP-2 Microparticles
[0073]A BMP-2 microparticle is prepared by mixing a first solution containing BMP-2 protein with a second, precipitating solution.
[0074]The first solution includes rhBMP-2 at a concentration of 0.1 to 2 mg / ml and is at pH 4.5. The second solution is formulated so that upon being added at one-tenth volume (relative to the volume of the first solution), the BMP-2 protein will precipitate and the final pH of the combined solution is pH 7.4. The second solution includes 1120 mM NaCl, 300 mM NaHCO3, 11 mM NaH2PO4, 5.5 mM Na2SO4, 40 mM NaOH and purified water.
[0075]The precipitation is performed by adding 10% (volume / volume) of a precipitating solution which contains both physiological salt and pH adjustment. The precipitation is performed in sterile tubes (15 or 50 ml), depending on the required volume. The filling of the 50 ml tube with BMP stock solution is limited to 30 ml to have enough space for the creation of a turbulent solution by using the vor...
example 2
Assessment of BMP-2 Microparticle Size
[0078]BMP-2 microparticle diameter was assessed using a MALVERN MASTERSIZER X® instrument (Malvern Instruments Ltd., Malvern, UK) with a sample cell designed for volumes of 4 to 5 ml. Mixing of a rhBMP-2 solution with a stock solution was used to generate physiologic salt concentrations.
[0079]The range in particle diameter in populations of microparticles subjected to equilibration for various lengths of time is shown in FIGS. 1-3. FIG. 1 shows the size distribution obtained in two batches microparticles equilibrated for less than ten minutes. Most particles are smaller then 10 μm in diameter, with a peak detected for microparticles of 5 μm to 6 μm in diameter.
[0080]FIG. 2 shows the size distribution obtained for three batches of microparticles equilibrated for between 10 and 110 minutes. While 6 μm to 9 μm are the most common size ranges, a higher number of microparticles with diameters greater than 10 μm are detected as compared to the distrib...
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