Cand. Sci. (bio) Petrusha Yu.Yu.

Zaporizhzhya National University, Ukraine

RESEARCH OF TOXIC ACTION OF S-PYRIDINE-MERCAPTOACIDS DERIVATIVES

INTRODUCTION. Pyridine chemistry for the last decades grew into one of the widest division of chemistry of heterocyclic substances. The heterocyclic system of pyridine is basis of many medicinal facilities that have a wide spectrum of pharmacological action. Substances, that show an antioxidant, antimicrobial, fungicide and other types of activity, were found among them. The possibility of using pyridinethyoles to receive the preparations with a cardioactive, analgetic, neurotropic action is especially valuable. Well-known facilities are on the basis of nicotinic and isonicotinic acids.

Clearly the practical value of substances of the indicated row does not finish on that examples and is not exposed yet. That’s why the pyridine system nowadays pretends on an intent attention of researchers. So the purposeful organic synthesis is the most perspective way to develop the pyridine chemistry. The research of biologically active substances in the rows of insufficiently known
S-pyridinederivates is important and has the theoretical and practical value [1, 2].

The introduction of biologically active substances, including regulators (stimulators) of height and development of plants (RPG) is perspective in a nowadays conditions. Using these preparations helps us to realize better the genetic possibilities, to rise the plant firmness against the stress factors of biotic and abiotic nature and at last to increase the harvest and to improve its quality [3]. Synthetic RPG from the derivative nitrous heterocyclic group, such as pyridine, are perspective to use in agriculture [4].

Results of researches of the last years [5, 6] show that combination of nitrogen-containing heterocycle and mercaptocarbonic acids influence on strengthening of biological action or appearance of new effects. Therefore a search of new bioactive substances containing in the molecule a heterocycle and a deputy with high antioxidant properties such as succinic acid or aminoacid L-cysteine is perspective.

That’s why on the basis of those studies [7] the di-Na-salt of 2-(pyridine-4-iltio)succinic acid and dihydrochloride S-(pyridine-4-il)-L-cysteine were selected for the deep research.

RESEARCH MATERIALS AND METHODS. Structure of the synthesized substances is confirmed by data of PMR-, ІR-spectroscopy and element analysis, and the cleanness – by the method of thin-layer chromatography (TLC).

The sharp toxicity was studied on white intact adults bisexual mouse weighing 20±3,0 g by means of Prozorovsky V. B. tabular express-method to determinate middle effective measures of influence on biological objects [8.

Researches of antibacterial activity of substance were conducted in vitro on methodology of the serial double breeding in a liquid nourishing environment (broth of Hоttingеr) [9]. Experiments executed on the 4 strain of bacteria, from that 2 cultures were gram-positive (Bacillus subtilis, Staphylococcus aureus), and other 2 – gram-negative (Escherichia coli, Pseudomonas aeruginosa). The estimation of results was carried out depending on intensity by oppressions of height of that or other culture of bacteria by a substance with a maximal concentration 500 mcg/ml.

The investigation of synthesized substance influence on a fission and cell growth (cytotoxicity) was done on a root test on the sprouts of Cucumis sp. (during experience used the cucumbers of sort "Competitor") [10]. Cytotoxicity of substance was estimated after reduction of the marked parameters in an experiment comparatively with control. Control (water) indexes are taken for a zero.

RESULTS AND DISCUSSION. The studies of sharp toxicity proved that di-Na-salt of 2-(pyridine-4-iltio)succinic acid is more or less safe according to the classification of Sidorov at intra-abdominal introduction to the wide range of doses. Animals were put the dose 4000 mg/kg and they were alive and active through 12, 24 hours and on a 14 day. Reflex activity, breathing depth and frequency of mice was not damaged. The reception of water, meal, egestion were not changed. Weight was not changed in comparison with the control group of animals. LD₅₀ of di-Na-salt of 2-(pyridine-4-iltio)succinic acid presents 4960±66 mg/kg.

The studies of sharp toxicity proved that dihydrochloride S-(pyridine-4-il)-L-cysteine is low-toxic according to the classification of Sidorov at intra-abdominal introduction. LD₅₀ of dihydrochloride S-(pyridine-4-il)-L-cysteine presents 832±81 mg/kg.

Research of antibacterial and cytotoxic action of di-Na-salt of 2-(pyridine-4-iltio)succinic acid confirmed the absence of toxic effects. It is found out that investigated substance does not have antibacterial activity at concentrations 500 mcg/ml. Cytotoxic action of di-Na-salt of 2-(pyridine-4-iltio)succinic acid was not marked, but even for concentrations 500 mcg/ml this substance stimulates fission and cell growth of sprouts of Cucumis sativus (figure 1).

%

hypocotyl length

length of the main root

length of a zone of lateral roots

quantity of lateral roots

Concentration, mkg/ml

Figure 1. Influence of di-Na-salt of 2-(pyridine-4-iltio)succinic acid

on a fission and cell growth of sprouts of Cucumis sp.

Research of antibacterial action of dihydrochloride S-(pyridine-4-il)-L-cysteine confirmed the absence of toxic effects. It is found out that investigated substance does not have antibacterial activity at concentrations 500 mcg/ml.

The research of the efficiency of dihydrochloride S-(pyridine-4-il)-L-cysteine (table 1) shows that the offered substance has a considerable grow-stimulating activity. It has ability to increase the length of main root and the amount of lateral roots of cucumber, that promotes the quick growing of leaves and productivity and viability of plants.

Table 1

Indexes of height stimulation of cucumbers under the influence of
dihydrochloride S-(pyridine-4-il)-L-cysteine

Parameters	Concentration, mcg/ml					Control (distilled water)
	1	5	20	100	500
Length of main root, mm	53,4±1,6*	47,6±2,1	48,6±2,2	53,4±2,0*	31,8±1,0	41,9±2,0
Length of height zone of lateral roots, mm	21,5±1,2*	17,8±0,9	17,1±1,1	20,7±1,0*	15,0±0,6	14,9±0,8
Amount of lateral roots	10,3±0,4	10,5±0,6	9,6±0,7	10,9±0,5	7,5±0,5	9,0±0,3

Primark: Р<0,05; * − concerning control. Control (the distilled water) it is accepted for zero.

CONCLUSIONS
1. It was found out that di-Na-salt of 2-(pyridine-4-iltio)succinic acid is relatively safe at intra-abdominal introduction to the wide range of doses (her LD₅₀ = 4960 mg/kg). Research of its antibacterial and cytotoxic action confirmed absence of toxic effects.

2. Di-Na-salt of 2-(pyridine-4-iltio)succinic acid is untoxic and can come forward as a substance for further researches to create new effective pharmaceutical preparations on its basis.

3. Dihydrochloride S-(pyridine-4-il)-L-cysteine is capable to increase the length of the main root and quantity of lateral roots of sprouts of gourd family that promotes more faster development of gypocotile and leaves and increases the productivity and viability of the plants.

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