Combination therapy comprising a polyunsaturated ketone and a secosteroid
A compound and composition technology, applied in the field of pharmaceutical compositions, can solve the problems that patients do not want to be seen, psoriasis is easily damaged, etc.
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Embodiment 1
[0154] The following compounds were used in the experiments:
[0155]
[0156] Co-Therapy Compound A and Calcipotriol:
[0157] method:
[0158] Cell culture:
[0159] At 37°C, 5% CO 2 The spontaneously immortalized non-tumorigenic dermal keratinocyte cell line HaCaT was maintained in a humidified atmosphere of 5% (v / v) FBS, 0.3mg / ml glutamine and 0.1mg / ml gentamicin in DMEM. Subculture with trypsin-EDTA in a split ratio of 1:3-1:4 every 3-4 days to ensure actively proliferating cells.
[0160] Resazurin assay:
[0161] Cells were seeded at a density of 2500 cells per well in 96-well plates in complete supplemented medium. After 72 hours of culture, cells were serum starved overnight in 0.25% FBS / DMEM to stop proliferation, synchronize and increase cell sensitivity to treatment. On day 4, cells were treated with the cPLA2α inhibitor compound A and the vitamin D analog calcipotriol hydrate (SigmaAldrich #C4369) and incubated at 37°C, 5% CO 2 Incubate for 2 hours in a...
Embodiment 2
[0168] Co-therapy with betamethasone and calcipotriol
[0169] method:
[0170] Cell culture:
[0171] At 37°C, 5% CO 2 The spontaneously immortalized non-tumorigenic dermal keratinocyte cell line HaCaT was maintained in a humidified atmosphere of 5% (v / v) FBS, 0.3mg / ml glutamine and 0.1mg / ml gentamicin in DMEM. Subculture with trypsin-EDTA in a split ratio of 1:3-1:4 every 3-4 days to ensure actively proliferating cells.
[0172] Resazurin assay:
[0173] Cells were seeded at a density of 2500 cells per well in 96-well plates in complete supplemented medium. After 72 hours of culture, cells were serum starved overnight in 0.25% FBS / DMEM to stop proliferation, synchronize cells and increase cell sensitivity to treatment. On day 4, cells were treated for 24 hours with the corticosteroid betamethasone 17,21-dipropionate (Sigma Aldrich #B1152) and the vitamin D analog calcipotriol hydrate (Sigma Aldrich #C4369). On day 5, resazurin was added according to the manufacturer's...
Embodiment 3
[0178] Compound A and calcipotriol showed a dose response on the viability of the immortalized keratinocyte cell line HaCat cells.
[0179] Cell culture:
[0180] At 37°C, 5% CO 2 The spontaneously immortalized non-tumorigenic dermal keratinocyte cell line HaCaT was maintained in a humidified atmosphere of 5% (v / v) FBS, 0.3mg / ml glutamine and 0.1mg / ml gentamicin in DMEM. Subculture with trypsin-EDTA in a split ratio of 1:3-1:4 every 3-4 days to ensure actively proliferating cells.
[0181] Resazurin assay:
[0182] Cells were seeded at a density of 3000 cells per well in 96-well plates in complete supplemented medium. After 48-72 hours in culture, cells were serum starved overnight in 0.25% FBS / DMEM to stop proliferation, synchronize cells and increase cell sensitivity to treatment. The next day, cells were treated with compound A or calcipotriol for 24 hours. The next day, resazurin was added according to the manufacturer's instructions (RnD Systems, UK) and incubated a...
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