Combination therapy comprising a polyunsaturated ketone and a folic acid partner
A composition and compound technology, which can be applied in the directions of medical preparations, drug combinations, and pharmaceutical formulations containing active ingredients, and can solve the problems that patients do not want to be seen by others, and that psoriasis is easily damaged.
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Embodiment 1
[0139] The following compounds were used in the experiments:
[0140]
[0141] Co-Therapy Compound A1 and Methotrexate:
[0142] method:
[0143] Cell culture:
[0144] 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.
[0146] 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 A1 and the folic acid analog methotrexate hydrate (MTX) (SigmaAldrich #A6770) and incubated at 37 °C, 5% CO 2 After incubatio...
Embodiment 2
[0151] PBMC isolation and experiments :
[0152] At St. Olavs Hospital HF, the Bloodbank recruits blood from healthy donors. Using SepMate as recommended by STEMCELLTechnology TM Separation tubes are separated from peripheral blood mononuclear cells (PBMC) with LymphoPrep density gradient medium. For experiments, 1x10 6 Cells / well / 1 mL RPMI medium supplemented with 5% FBS, 0.3 mg / ml glutamine and 0.1 mg / ml gentamicin, with or without inhibitor compound A1, methotrexate hydrate (MTX) ( Sigma-Aldrich, #A6770) or a combination of two inhibitors, before the addition of lipopolysaccharide as a potent inducer of inflammation (γ-irradiated LPS from E. coli 026:B6, Sigma-Aldrich, #L2654). Treatment (72 hours, 37°C, 5% CO 2 ), the cell suspension was centrifuged to separate the supernatant from the cell fraction. Samples were stored at -80°C until analysis.
[0153] ELISA detection of PGE2, TNF and IL1-β
[0154] Cell supernatant samples were analyzed by enzyme-linked immuno...
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