Complex of n,n-diethyldithiocarbamate sodium with 2-ethyl-6-methyl-3-hydroxypyridine eliciting anti-allergic activity

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to a new complex of N,N-diethyldithiocarbamate with 2-ethyl-6-methyl-3-hydroxypyridine of the formula: . This complex elicits an anti-allergic activity that allows it using for carrying out the pharmacological control of allergic response based on inhibition of function of allergy target-cells.

EFFECT: valuable medicinal properties of complex.

1 cl, 1 tbl, 3 ex

The invention relates to new chemical compounds with biological activity, and can be used to create pharmacological agents with anti-allergic action.

Current views on the mechanism of allergic reaction suggest that one of the most important factors in the formation of the allergic process is the involvement of mast cells of the mucous membranes and connective tissue, and basophilic leukocytes in peripheral blood (Gushchin I.S. "Allergic inflammation and its pharmacological control), Farmers print, 1998, p.52-58) the Connection of the allergen with the antibody on the surface of these cells triggers the secretion of mediators that cause pathophysiological changes (smooth muscle contraction, increased vascular permeability, mucus hypersecretion of mucous glands)that form the basis of clinical manifestations of Allergy.

It is established that mast cells (TC) and basophilic leukocytes, which are the main target cells of allergies are involved in allergic reactions by activating their function. Therefore, it becomes all the more urgent directed search for new compounds that could have an impact on the functional state of the target cells allergies.

Known PR is parathas kromoglicieva acid and nedocromil sodium (Gushchin I.S. "Allergic inflammation and its pharmacological control), Farmers print, 1998, str-222), anti-allergic activity of which is associated with a stabilizing action on target cells allergies. However, this effect is rather weak and is manifested in the experimental conditions with high concentrations of these compounds that have no therapeutic value.

Thus, the technical result from the implementation of the present invention is to provide new chemical compounds having anti-allergic action due to inhibition of the function of target cells allergies.

This technical result is achieved by a complex of N,N-diethyldithiocarbamate sodium 2-ethyl-6-methyl-3-hydroxypyridine General formula:

having antiallergic activity.

Compound (adduct) N,N-diethyldithiocarbamate sodium 2-ethyl-6-methyl-3-hydroxypyridine is a new compound and can be obtained by complexation of N,N-diethyldithiocarbamate sodium 2-ethyl-6-methyl-3-hydroxypyridine in equimolar proportions, carried out in anhydrous medium in the presence of an aprotic solvent.

The formation of complex compounds is due to the donor-acceptor bond, due to the Oh electroreceptors properties of substituted 3-hydroxypyridine and high electronegativity of sulfur atoms in N,N-diethyldithiocarbamate sodium.

In accordance with the invention found that the complex compound (adduct) N,N-diethyldithiocarbamate sodium 2-ethyl-b-methyl-3-hydroxypyridine (CDC) can affect allergic process by suppressing the activation functions of mast cells and basophilic leukocytes and are thus anti-allergic activity.

The complex compound in accordance with the present invention was prepared as follows.

In a three-neck flask equipped with stirrer, thermometer and a fridge, loads of 3.14 g (0,02 Mol) of N,N-diethyldithiocarbamate sodium, poured 60 ml of absolute isopropyl alcohol and 2 ml of acetone, and then added 2.76 g (0,02 Mol) of 3-hydroxy-6-methyl-2-ethylpyridine with constant stirring until complete dissolution of the components. Next, the resulting solution was stirred at 40° C for 3 hours, then filtered and the solvent is distilled off in vacuum. The precipitate is recrystallized from absolute isopropyl alcohol. Get 4,25 g of white crystalline product

N,N-diethyldithiocarbamate 3-hydroxy-6-methyl-2-ethylpyridine sodium.

T_PL=130-133° C. Soluble in water, in alcohol.

Found, %: C 43,11; N To 7.61; N 7,56; S 17,61.

Calculated, %: 43,07; N 7,51; N 7,73; S 17,69.

IR spectrum^-1: 1385 (S=C<); 1610 (C=C, arene.); 3380 (HE). Study of the biological activity of the compounds according to the invention is carried out using a method of determining the release of histamine from mast cells of rats, the method of release of histamine in vitro from basophils in the peripheral blood of man and the passive cutaneous anaphylaxis in vivo. Preliminary studies suggest the possible exception of the studied compounds to interact with the reaction determination of histamine and cause within the tested concentrations, the release of histamine from mast cells and basophils.

The physiological activity of the described compounds shown in examples 1-3.

Example 1. Determination of the release of histamine from mast cells of rats.

In tests using outbred white rats-males weighing 250-300 g

Fat cells (TC) rats separated from the abdominal and thoracic cavities by centrifugation on density gradient Pichola. Contents TC in a dedicated suspension of cells was 85-95%. In a series of test tubes containing 2 ml of saline incubate solution, pre-make a test compound in various concentrations. In the control test tube test connection does not contribute. The tube is maintained at a temperature of 37° for

3 minutes, after which they are 5 µl suspension of TC and incubated for 20 minutes After incubation the cells add standard altoconsky visualeditor (substance 48/80), which causes rapid activation of the LC, to final concentrations: 1 μg/ml, 0.5 μg/ml and 0.2 µg/ml is eakly activation of the LC stopped by transferring the tubes on ice, after that the cells are precipitated by centrifugation at 400 g and 4° C for 10 min. the Supernatant poured into a new tube, the cells are lysed by the addition to precipitation, 2 ml of distilled water and shaken on a Vortex mixer. In the supernatant and sediment samples to determine the content of histamine spectrofluorometrically method.

The research results presented in figure 1 show that CDC markedly inhibits the secretion of histamine from TC, caused by substance 48/80, and at concentrations of 10 μm is observed 100% inhibitory effect.

Example 2. Determination of the release of histamine from basophilic leukocytes in human peripheral blood.

For research use blood of patients with pollinosis with a heightened sensitivity to the pollen of grasses and trees installed along with pre-made skin diagnosticheskii samples with the allergen. Otbiraut material from patients aged 15 to 34 years with moderate and severe symptoms of pollinosis in the form of rhinitis, conjunctivitis and bronchial asthma.

From the blood of patients by well-known methods (Gushchin I.S., Leskov VP, Kitaeva VG, Polsacie Century, W. Immunology, 1982, No. 4, p.67-71) receive a suspension of mononuclear cells containing 2-4% basophils, and prepare samples with the test compounds at concentrations of 10 μm, 100 μm and 1 mm. Allergens (grass pollen, is ereview or house dust) used in the concentration range from 0.1 to 10.0 PNU/m1.

As a control using samples without addition of test compounds.

Prepared samples incubated for 40 minutes at a temperature of 37° With, then make allergen and continue incubation of the cells in the same conditions for another 40 minutes the Release of histamine determine microspectrophotometric method in sedimentary portions without prior extraction and expressed in percentage of the total content of histamine in the serving cell.

Discovered that CDC suppresses the activation of target cells human basophils.

The data in table 1 suggests that the analyzed connection depending on concentration has a clear depressing effect.

Example 4. The study of passive cutaneous anaphylaxis (PKA).

Studies carried out on outbred rats male body weight 210-240,

The reaction PKA reproduce previously described method [Levin CENTURIES, Vaz N. M. Int. Arch. Allergy. 1970, r.39-196].

The analyzed connection prior to throw in the quantity of 100 mg in 10 ml saline and administered to the animals intraperitoneally in a dose of 100 mg per 1 kg of body weight of the test animal 30 minutes before intravenous injection of a resolving dose of antigen (ovalbumin). As a control use animals that are injected with 0.9% NaCl solution.

The results are judged by the threshold cutaneous reactions, what idivi on the inner side of the skin. For threshold reactions take the greatest dilution (titer) antialbumin serum, which occurs the staining of the skin.

The graphs presented in figure 2, illustrate the reduction of the titer of the reaction PKA when playing on the rats that received the described connection.

Thus, a new connection has antiallergic activity and can be used in the development of new drugs for the pharmacological control of allergic reactions and, therefore, is a potential anti-allergic agent.

Complex of N,N-diethyldithiocarbamate sodium 2-ethyl-6-methyl-3-hydroxypyridine General formula:

having antiallergic activity.