CS276923B6 - Immunomodulatory means - Google Patents

Abstract

The solution relates to immunomodulatory agents which have as an active ingredient in their molecule ammonium bromides of the above formula and contain in the molecule one or two long alkyl chains with a number of carbon atoms of 11 to 14 and which, depending on the concentration, exhibit either an immunosuppressive or immunostimulatory effect. Due to these effects, they can be used, among other things, in human and veterinary medicine and pharmacy.

Description

t

I 1 EN 276923 B6

The present invention relates to compositions having immunomodulatory effects, the active ingredient of which is an amphiphilic compound of the group of organic ammonium salts of the formula R @ + + N - R @ 1 Br @ - wherein R is a carbon chain linear or branched alkyl chain of from 11 to 14, R1 is CH3, Cioh21 C14H29 and combinations.

Organic ammonium salts containing at least one long alkyl chain in their molecule are amphiphilic compounds which have been used for a long time in various fields of human activity mainly as disinfectants in human and veterinary practice. they also have significant biological activities. Recently (Ashman R. B., Blanden R.V., Ninham B.W., Evans D.F., Immunol. Today 7, 278 (1986)), some simple ammonium salts containing molecular or two long alkyl chains interact with cells of the immune system. Their observed activity was higher than cyclosporin A, the most potent immunospuppressive drug currently used. Dioctadecyldimethylammonium bromide, in turn, has immuno-malignant properties (Hilgers, L.T., Snippe, H., Jansze, M., Wilson, J., N., Cell. Immunol. 90, 14 (1985)).

Immunomodulatory agents can increase (immunostimulators) but destroy (immunospures) the activity of the immune system or some of its functions. Many immunomodulators usually have both of these properties. Which of them will depend depends on the concentration of the immunomodulator, the conditions and method of its application, the experimental model and the test function of the immune system (Dean, H., Luster, I., Munson, AE, Amos, HA eds. Immunotoxicity and Immunopharmacology, New York, Raven Press 1983, p 511).

One of the most important mechanisms of natural immunity is phagocytosis. Its role is to capture, inactivate and decompose foreign particles, including pathogenic microorganisms, foreign self-antigenic and functionally altered cells (Ferencich., Folia Fac. Med. Univ Comenianae Bratisl., 24, 9 (1986). professionally phagocytes (neutrophils, eosinophils, monocytes, and tissue macrophages), therefore, these cells are useful for testing the effects of various immunomodulators, and for the active compounds of the invention, the previously known immunosuppressive and immunostimulatory effects of the metabolic activity of neutrophils isolated from Test substances at concentrations of > 4mol / dm 3 were found to have an inhibitory effect on the INT-reductase activity of human peripheral neutrophils, while lower concentrations had no significant effect, and concentrations of > 10-7 mol / dm 3 of the test substances significantly stimulated superoxide formation. -v conc concentrations> 10 4 mol / dm 3 had a lysosome membrane-labile effect, resulting in increased extracellular secretion

Claims (1)

3 CS 276923 B6 53 (1979). Test compounds at concentrations of 10 3 to 10 7 mol / dm 3 significantly increased the amount of granulocyte-released enzyme activities into the incubation medium. Compound c [mol / dm3] Increase of BDG enzyme [%] MPO activity L (1-metododecyl) tri-control 16 13 13 Methylammonium bromide Ix10 3 45 24 60 Didodecyldimethyl Ix10 6 6 o 25 11 31 Mium bromide X 10; 60 28 33 1X10 “'18 20 28 PATENT REQUIREMENTS Immunomodulatory agents, characterized in that they contain as active compounds amphiphilic compounds of the general formula CH, N" Rx Br CH- wherein R is a linear or branched alkyl chain with a carbon atom of 11 to 14, R 1 is CH3, C10H21 to C14H29 and their combinations The endpoint of CS 276923 2 by lysosomal enzymes from neutrophils suggests that the compounds of interest affect the metabolic activation of phagocytes, dynamic equilibrium in superoxide production and high concentrations as well as The examples illustrate but do not limit the scope of the types of immunomodulators according to the invention: Example 1 Preference blood was drawn from healthy volunteers into heparin (20 µ / cm 3) and granulocytes sadimentacious Dextran were isolated therefrom T-500 (Boyum A., Scand. J. Immunol. 5, Suppl. 9 (1976)) Only a granulocyte suspension containing at least 90% neutrophils was used. Pure granulocytes were resuspended in phosphate buffered saline (PBS) containing 5 µl of heparin per cm 3 and 0.1% glucose (PBSHG). Their INT-reductase activity as well as the production of superoxide are determined by micromethod in microtiter plates (Ferencik M., Kotulova, Masler, L., Sandula, J., Pruzinec, P., Bratislava, 89,424 (1988)). These activities were found in both unstimulated granulocytes and in granulocytes stimulated by opsonized zymosane (ZO) and phorbol myristate acetate (PMA) in the absence (control) and in the presence of test substances. All of the investigated substances in high concentrations (> 10-4 mol / dm 3) reduced the ability of both non-stimulated and stimulated granulocytes to reduce INT (suppressive effect) by an average of 30-90%. For example, (1-methyldodecyl) trimethylammonium bromide inhibited the granulocyte immunological response at c = 1Q-3 mol / dm 3 to 45%; didodecyldimethylammonium bromide at c = 10 "3 mol / dm 3 to 35%. Example 2 The procedure was the same as in Example 1, except that superoxide production was monitored. Unstimulated | granulocytes were observed at 10 4 4 to 10 7 7 mol / dm3 stimulating effect: (1-methyldodecyl) trimethylammonium bromide at c = 10 ”7 mol / dm3 stabilization by 240%, at c = 10” 4 mol / dm3 stimulation by 135%, · didode-cyldimethylammonium bromide at c = 10 “5 mol / dm3 stimulation by 237%, at c = 10” 4 mol / dm3 stimulation by 60%. The concentration of c = 10 3 mol / dm 3 for all substances inhibited superoxide formation. Example 3 Granulocytes were isolated from clinically healthy volunteers as described in Example 1. They were adjusted to a concentration of 077 / cm 3 in PBSHG and incubated for 60 min at 37 ° C in pure PBSHG or with stimulants (ZO, PMA) in presence (control) and in the presence of test substances. After incubation with sacrifice, the supernatants separated from the granulocyte sediment and found the percentage of extracellularly released lysosomal enzymes - β-D-glucuronidase (BDG), myeloperoxidase (MPO) and lysozyme (L) (Ferenčík M., Beláň J., Bratisl. Lek Letters 71,