Combination therapy for treatment of mastocytosis
A technology of mast cells and cells, which can be applied in drug combinations, biochemical equipment and methods, microbial measurement/testing, etc., and can solve problems such as side effects
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Embodiment 1
[0160]Example 1. Treatment of mutant KIT mast cell lines with Compound A or Compound B inhibits cell proliferation and KIT phosphorylation in mastocytosis cell lines.
[0161] The studies performed showed that treatment with Compound A or Compound B inhibited cell proliferation of mutant KIT HMC1.1KIT V560G and HMC1.2 KIT V560G / D816V mast cell lines. Assays were performed in 96-well plates seeded with 10,000 cells per well. Cells were treated with vehicle control, Compound A or Compound B at various concentrations, allowed to grow for 72 hours, and then assessed for cell proliferation.
[0162] Figure 1A is a graph showing the relative percentage of cell proliferation determined for various concentrations of Compound A. Treatment with Compound A inhibited cell proliferation and growth of HMC1.1 V560G and HMC1.2 V560G / D816V mast cell lines with IC50 values of 2.6nM and 97nM, respectively.
[0163] Figure 1B is a graph showing the relative percentage of cell proliferation...
Embodiment 2
[0164] Example 2. Treatment with the combination of Compound A and trametinib induces apoptosis in mastocytosis cell lines.
[0165] The studies performed showed that combined treatment with compound A and the MEK inhibitor trametinib induced apoptosis in the HMC1.2 KITV560G / D816V mastocytosis cell line. Assays were performed in 96-well plates seeded with 10,000 cells per well. Cells were treated with vehicle control, Compound A, Trametinib or combinations thereof at various concentrations and grown for 24 hours. Apoptosis was assessed by measuring caspase 3 / 7 activity.
[0166] Figure 2A is a graph showing the relative percentage of apoptosis determined for various treatments. Figure 2B is a matrix based on the combination index (CI) method described by Chou and Talalay (1984) and the synergy diagram of computer software by Chou and Martin (2005). CI1 indicates antagonism. Combination treatment with compound A and the MEK inhibitor trametinib unexpectedly showed a stro...
Embodiment 3
[0167] Example 3. Treatment with the combination of Compound B and trametinib induces apoptosis in mastocytosis cell lines.
[0168] Studies were also performed showing that combined treatment with compound B and trametinib induced apoptosis in the HMC1.2KIT V560G / D816V mastocytosis cell line. Assays were performed as described in Example 2. Apoptosis was assessed by measuring caspase 3 / 7 activity.
[0169] Figure 3A is a graph showing the relative percentage of apoptosis determined for various treatments. Figure 3B is a matrix based on the combination index (CI) method described by Chou and Talalay (1984) and the synergy diagram of computer software by Chou and Martin (2005). CI1 indicates antagonism. Combination treatment with compound B and the MEK inhibitor trametinib unexpectedly showed a strong synergy to induce apoptosis in HMC1.2KIT V560G / D816V cells. Figure 3C is the combination index graph of CI, indicating that the combination of compound B and trametinib ha...
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