CS276929B6 - Immunomodulatory means - Google Patents

Abstract

The solution relates to immunomodulatory agents which have as active ingredient amine oxides of the general formula mentioned and contain in the molecule one long alkyl chain with a number of carbon atoms of 11 to 14 or a long chain and a heterocycloalkane of the formula mentioned and which, depending on the concentration, exhibit either an immunosuppressive or immunostimulating effect. Due to these effects, they can be used, among others, in human and veterinary medicine and pharmacy.

Description

The invention relates to compositions with immunomodulatory effects, the active ingredient of which are amphiphilic compounds from the group of amine oxides of the general formula

CHo »-4 - oI ch ₃

wherein R represents a straight or branched alkyl chain having 11 to 14 carbon atoms, and X = (CH ₂ ) _n where n = 4, 5, 6 or (ch ₂ ch ₂ ) 0.

Amine oxides containing at least one long alkyl chain in their molecule are amphiphilic compounds which have been used for a long time in various branches of human activity. They are known, in addition to their detergent effect, they also have significant biological activities. It has recently been found (Ashman R. B., Blanden R. V., Ninham B. W., Evan D. F., Immunol. Today 7, 278 (1986)) that certain structural analogs of amine oxides, such as e.g. simple ammonium salts containing one or two long alkyl chains in their molecule interact with cells of the immune system. Their observed activity was higher than cyclosporin A, the most effective immunosuppressive drug currently in use. Dioctadecyldimethylammonium bromide, in turn, has immunostimulatory properties (Hilgers L. A. T., Snippe H., Jansze M., Willers J. N., Cell. Immunol. 90, 14 (1985)). However, for amphiphilic compounds containing an amine oxide group in their molecule, no knowledge is currently known as to whether and how they affect the immune system.

Immunomodulatory agents can increase (immunostimulators) or decrease (immunospursives) the activity of the immune system or some of its functions. Many immunomodulators usually have both of these properties. Which of them 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 JH, Luster I.. I., Munson AE, Amos HA eds .: Immunotoxicology and Immunopharmacology. New York , Raven Press 1983, p. 511).

One of the most important mechanisms of innate immunity is phagocytosis. Its role is to capture, inactivate and decompose foreign particles, including pathogenic microorganisms, foreign and self-antigenically and functionally altered cells (Ferenčík M., Folia Fac. Med. Univ. Comenianae Bratislava 24, 9 (1986)). neutrophils, eosinophils, monocytes and tissue macrophages) These cells are therefore preferably used to test the effect of vigorous immunomodulators.

The active compounds of the invention have been found to have unknown immunosuppressive and immunostimulatory effects on the metabolic activities of neutrophils isolated from human peripheral blood. Test substances at concentrations> 10 ^-4 mol / dm ³ were found to have an inhibitory effect on the INT-reductase activity of human peripheral neutrophils, while lower concentrations had no demonstrable effect. Concentrations> I0 “ ⁷ mol / dm ^{3 of} test substances significantly stimulated superoxide production. The monitored substances at concentrations> 10 ” ⁴ mol / dm ³ had a labilizing effect on lysosomal membranes, which was manifested by increased extracellular secretion of lysosomal enzymes from neutrophils.

It follows from the above that the monitored substances by their effect interfere with the metabolic activation of phagocytes, the dynamic balance in the production of superoxide and in high concentrations also in the function of lysosomes in neutrophils. They therefore have a demonstrable immunomodulatory effect on phagocytic function.

The examples illustrate but do not limit the scope of the types of immunomodulators of the invention.

Example 1

Peripheral blood was collected from heparin (20 J / cm ³ ) from healthy volunteers, and granulocytes were isolated from it by sedimentation with Dextran T-500 (Boyum A., Scand. J. Immunol. 5, Suppl. 5, 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 heparin per cm 3} and 0.1% glucose (PBSHG). Their INT-reductase activity as well as superoxide production was determined by micromethoding on microtiter plates (Ferenčík M., Kotulová D., Masler L., Šandula J., Pružinec P., Bratisl. Lek. Listy 89, 424 (1988)). These activities were detected in unstimulated granulocytes as well as in granulocytes stimulated with opsonized zymosan (ZO) and phorbol myristate acetate (PMA) in the absence (control) and in the presence of test substances. All test substances at high concentrations (> 10 ^-4 mol / dm ³ ) reduced the ability of unstimulated and stimulated granulocytes to reduce INT (suppressive effect) by an average of 30 to 90%. for example, it inhibited the immune response of ³ to 47% at c = 10 ^-3 mol / dm ³ of

%.

Example 2

The procedure was the same as in Example 1, except that superoxide production was monitored. In the case of unstimulated granulocytes, a ^{stimulatory effect was observed at concentrations of 10 -4} to 10 mol / dm ³ : for example (1-methyldodecyl) dimethylamine oxide at ^{231 = 7} mol / dm ³ stimulation by 231%, at c = 10 ^-4 mol / dm ³ stimulation by 50%; The concentration of c = 10 ” ³ mol / dm ³ for all substances inhibited the formation of superoxide.

Example 3

Granulocytes were isolated from clinically healthy volunteers as described in Example 1. They were adjusted to a concentration of 10 ⁷ / cm ³ in PBSHG and incubated for 10 min at 37 ° in pure PBSHG or together with stimulants (ZO, PMA) and in the absence (control) and in the presence of test substances. After incubation, the supernatants were separated from the granulocyte sediment by centrifugation and the percentage of extracellularly released lysosomal enzymes - β-D-glucuronidase (BDG), myeloperoxidase (MPO) of granulocytes at c = 10 ^-4 mol / dm /

/ and lysozyme (L) (Ferenčík Μ., Beláň J., Bratisl. lek. listy 71, (1979)). Test compounds in concentrations of 10 ³ to

10 ⁷ mol / dm ³ significantly increased the amount of enzyme activities released from granulocytes into the incubation medium.

Compound ^C o [mol / dm ^J ] Increase in enzyme activities BDG [%] MPO L control 4 7 5 (1-methyldodecyl) dimethylamine oxide ^a IxlO ³ 28 44 50 IxlO “ ⁷ 25 11 31 4-tridecylmorpholine N-oxide ixio; 88 60 86 2xl0 ” ⁷ 6 12 . 12 1-dodecylpyrrolidine N-oxide 5xio; 49 60 70 2xl0 ⁷ 6 13 9 1-tridecylpiperidine N-oxide 1X10 ° 90 57 61 2xl0 ⁷ 14 19 14 1-tridecyl perhydroa- zepine N-oxide 2x10 ' 31 36 35 4x10 ° 5 13 8

^and incubation for 60 min, control 16% (BDG), 13% (MPO), 13% (L)

Claims (1)

PATENT CLAIMS Immunomodulatory compositions, characterized in that they contain amphiphilic substances of the general formula as active compounds Γ ³ R --- N ⁺ --- OR --- ⁺ NX ch ₃ “O where R represents a linear or branched alkyl chain with the number of carbon atoms 11 to 14, and X = (CH ₂ ) _n where n = 4 to 6 and (CH ₂ CH ₂ ) ₂ O. End of document