Use of a bisamide derivative of malonic acid for treating allergic diseases and other diseases in humans and animals
A technology for allergic rhinitis, use, applied in the field of organic chemistry, which can solve the problems such as the absence of antagonists of type 3 and type 4 histamine receptors
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Embodiment 1
[0153] Example 1. Study on the Effect of Compound I on Type 3 Histamine Receptor Activity
[0154] Compound I was dissolved in DMSO to a concentration of 100 mM; the stock solution was then serially diluted with DMSO. The maximum starting concentration of substances is 100 μM. Effects were determined at 8 concentrations of test compound; each concentration was tested in duplicate. The cells used in the experiments were CHO cells expressing the human type 3 histamine receptor, which were incubated with compound I after pre-incubation with histamine. The activity of the receptor was determined by intracellular cAMP concentration using fluorescence spectroscopy (Lim, H.D. et al. (2005) J. Pharmacol. Exp. Ther., 314:1310-1321).
[0155] In this study, compound I was found to be an antagonist of the type 3 histamine receptor and the IC 50 = 20 μM.
Embodiment 2
[0156] Example 2. Study of the Effect of Compound I on the Binding of Radiolabeled Ligands to Histamine Receptor Type 3
[0157] Compound I was dissolved in DMSO to a concentration of 300 mM; the stock solution was then serially diluted with DMSO. The maximum starting concentration of substances is 300 μM. Effects were determined at 10 concentrations of test compound; each concentration was tested in duplicate. Cells used in the experiments were CHO cells expressing the human type 3 histamine receptor, which were incubated with compound I after pre-incubation with [3H]Nα-methylhistamine (1 nM). The activity of Compound I was determined by the concentration of displaced radiolabeled ligand (Lovenberg, T.W. et al. (1999), Mol. Pharmacol., 55:1101-1107).
[0158] In this study, compound I was found to bind to the type 3 histamine receptor with a Ki value of 2.7 μΜ.
Embodiment 3
[0159] Example 3. Study of the Effect of Compound I on the Binding of Radiolabeled Ligands to Histamine Receptor Type 4
[0160] Compound I was dissolved in DMSO to a concentration of 300 mM; the stock solution was then serially diluted with DMSO. The maximum starting concentration of substances is 300 μM. Effects were determined at 10 different concentrations of test compound; each concentration was tested in duplicate. The cells used in the experiment were HEK-293 cells expressing the human histamine receptor type 4, which were incubated with compound I after pre-incubation with [3H]histamine (10 nM). The activity of compound I was determined by the concentration of displaced radiolabeled ligand (Liu, C. et al. (2001), J. Pharmacol. Exp. Ther., 299:121-130).
[0161] In this study, compound I was found to bind to the type 4 histamine receptor with a Ki value of 16 μΜ.
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