Combination therapy for the treatment of immunoinflammatory disorders
A technology of immune factors and compositions, applied in allergic diseases, skin diseases, bone diseases, etc., can solve problems such as adverse side effects
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Embodiment 1
[0207] Example 1: Determination of pro-inflammatory cytokine-inhibitory activity
[0208] Compound dilution models were assayed for inhibition of IL-2 or TNFα as described below.
[0209] IL-2
[0210]Stimulation in the IL-2 was secreted from 100 μL of the diluted human leukocyte suspension contained in each well of a polystyrene 384-well plate (NalgeNunc). Various concentrations of each test compound were added at the time of stimulation. After 16-18 hours of incubation at 37°C in a humidified incubator, the plate was centrifuged and the supernatant was transferred to white opaque polystyrene 384 wells coated with anti-IL-2 antibody (PharMingen, #555051) plate (NalgeNunc, Maxisorb). After two hours of incubation, the plate (Tecan Power Washer 384) was washed with PBS containing 0.1% Tween 20 and another anti-IL-2 antibody labeled with biotin (Endogen, M600B) and streptavidin (PharMingen, #13047E)-coupled HRP was incubated for 1 hour. After the plate was washed with 0.1%...
Embodiment 2
[0217] Embodiment 2: the preparation of compound
[0218] Stock solutions containing NsIDI and group A enhancers were prepared at final concentrations between 0 and 40 μΜ in dimethyl sulfoxide (DMSO). Prepare plates containing dilutions of the above compound stock solutions. The template was sealed and stored at -20°C until use.
[0219] NsIDI and Group A enhancer stock solution
[0220] A stock solution containing cyclosporine A at a concentration of 1.2 mg / ml was prepared in DMSO. A stock solution of tacrolimus was prepared at a concentration of 0.04 mg / ml in DMSO.
[0221] A stock solution containing acyclovir was prepared at a concentration of 10 mg / ml in DMSO. A clotrimazole-containing stock solution was prepared at a concentration of 10 mg / ml in DMSO. Zinc-containing stock solutions were prepared at a concentration of 10 mg / ml in DMSO. A stock solution containing urea was prepared at a concentration of 10 mg / ml in DMSO. Oxybenzone-containing stock solutions were p...
Embodiment 3
[0224] Example 3: Combination of Tacrolimus and Acyclovir Reduces IL-2 Secretion in Vitro
[0225] After stimulation with phorbol-12-myristate-13-acetate and ionomycin, IL-2 secretion was determined by ELISA as described above. The effects of different concentrations of tacrolimus, acyclovir, and combinations of tacrolimus and acyclovir were compared to control wells stimulated without tacrolimus or acyclovir. The results of this experiment are shown in Table 3 below. The effects of individual drugs and combined drugs were expressed as percent inhibition of IL-2 secretion. The data below represent data from one experiment for individual drugs and combinations. Wells without serial numbers indicate artificial data that have been omitted.
[0226] table 3
[0227]
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