Combination composition comprising FGF-18 compound
A technology of FGF-18, 1.FGF-18, applied in the direction of drug combination, chemical instruments and methods, medical preparations containing active ingredients, etc.
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Embodiment 1-F
[0090] Example 1 - Combination of FGF-18 and IL-6 Inhibitors
[0091] method:
[0092] BaF3 / FGFR3c bioassay: 1x10 7 cells at 75cm 2 Inoculate 20 mL of assay medium in flasks for 24 h at 37 °C, 5% CO 2 , for the IL-3 starvation step. On the day of the experiment, 20,000 cells / well were seeded in 50 μL of assay medium in a 96-well plate containing 0.1, 1, 10, 100, 1000, and 10,000 ng / mL of CNTO328 or 0.001, 0.01, 0.1, 1 , 10 and 100 ng / mL of PMP6B6 with or without 100 ng / mL spfermin. As a control, cells were also cultured with 100 ng / mL of spfermin alone (positive control, CTR+ on the graph), BMM2 1 / 2200 or without any compound (both are negative controls, CTR- on the graph). All conditions were achieved with N=6. Cells at 37°C, 5% CO 2 After culturing for 2 days, the metabolic activity was measured with WST-1 reagent (Roche).
[0093] Primary human chondrocyte culture: After cell isolation, human chondrocytes were seeded at 14-18 million cells in 75cm 2 flasks and ...
Embodiment 2-F
[0105] Example 2 - Combination of FGF-18 and IL-6 receptor inhibitors
[0106] method:
[0107] BaF3 / FGFR3c bioassay: The same method and conditions as described in Example 1 were used. On the day of the test, 20,000 cells / well were inoculated in 50 μL of test medium in a 96-well plate containing 0.001, 0.01, 0.1, 1, 10 or 100 μg / mL of tocilizumab (Roche), And contain or not contain 100ng / mL spfermin. Cells cultured with (CTR+) or without (CTR-) spfermin and in excipients of the tocilizumab formulation (15 mM sodium phosphate, 0.5 mg / mL polysorbate 80, 50 mg / mL sucrose, pH 6, 5. Controls were achieved in the presence of a 1 / 200 dilution in culture medium to correspond to the highest tocilizumab concentration). All conditions were achieved with N=6. Cells at 37°C, 5% CO 2 After culturing for 2 days, the metabolic activity was measured with WST-1 reagent (Roche).
[0108] Primary human chondrocyte culture: The same method and conditions as described in Example 1 were u...
Embodiment 3-F
[0116] Example 3 - Combination of FGF-18 and NGF Inhibitors
[0117] method:
[0118] BaF3 / FGFR3c bioassay: The same method and conditions as described in Example 1 were used. On the day of the assay, 20,000 cells / well were seeded in 96-well plates in 50 μL of assay medium containing 0.01, 0.1, 1, 10, 100, or 1000 nM of tanezumab with or without 100ng / mL spfermin. Positive control (CTR+) was achieved by cells incubated with 100 ng / mL sprifermin in the absence of tanizumab. All conditions were achieved with N=6. Cells at 37°C, 5% CO 2 After culturing for 2 days, the metabolic activity was measured with WST-1 reagent (Roche).
[0119] result( Figure 7 ):
[0120] Tanizumab had no effect on cell proliferation and did not interfere with spruferin. Increases in tanizumab concentration did not affect cell proliferation and the O.D. remained at 0 when spfermin was not administered. In the presence of spfermin, BaF3 / FGFR3 cell proliferation was increased, resulting in ...
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