RU2365596C1 - Copolymers of diallylaminophosphonium salts with sulphur dioxide demonstrating anti-microbial activity - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: claimed invention relates to copolymers of diallylaminophosphonium salts with sulphur dioxide demonstrating antimicrobial activity with respect to a number of bacteria, as well as to yeast-like fungi and spores, and can be applied as antiseptic and disinfecting means. Claimed copolymers of diallylaminophosphonium salts with sulphur dioxide aree characterised by general formula

where A=Cl^- or BF₄ ^-. They are soluble in methanol, DMSO, DMFA or if A=Cl^- are soluble in water. They are obtained by copolymerisation of equimolar amounts of sulphur dioxide and diallylaminophosphonium salt, selected from tris(diethylamino)diallylaminophosphonium chloride or tris(diethylamino)diallylaminophosphonium tetrafluoroborate.

EFFECT: obtaining novel efficient and low-toxic compounds which do not cause corrosion of processed metals.

2 cl, 3 tbl, 4 ex

Description

The invention relates to copolymers of diallylaminophosphonium salts with sulfur dioxide, exhibiting antimicrobial activity, and may find application as antiseptic and disinfectant.

Descriptions of the claimed compounds, their properties and their use as antiseptic and disinfectants in the information sources were not found.

Analogs of the claimed compounds in biological activity are various antiseptics and disinfectants.

So, traditional chlorine-based products, such as chloramine B (sodium salt of N-chloramide of benzenesulfonic acid, are widely used for disinfection

C ₆ H ₅ SO ₂ NNaCl · 3H ₂ O) [Efimov K.M., Gembitsky P.A., Snezhko A.G. Polyguanides - a class of low-toxic long-acting disinfectants // Disinfection business, 2000, No. 4]. With moderate antimicrobial activity, this drug is highly toxic, irritates the skin and mucous membranes. In addition, it has a pronounced pungent odor and is corrosive.

Quaternary ammonium compounds are known as disinfectants, for example, the drug Alaminol (alkyldimethylbenzylammonium chloride) phosphate [Ibid.]. This drug also shows toxicity and has an irritating effect on the skin and mucous membranes, although more moderate in comparison with the above means. The disadvantage is its low activity against spore cultures of microorganisms.

The closest to the achieved effect analogue of the proposed solution are polyguanidines, for example phosphopag-polyhexamethylene guanidine phosphate [Ibid].

Phosphopagum has a high biocidal effect in relation to a number of microorganisms and belongs to the 4th class of toxicity when it is ingested by the skin and to the 3rd class when it is delivered through the stomach, which indicates the presence of low toxicity. However, there is no data on the activity of phosphopag in relation to Micrococcus luteus, Staphylococcus saprophyticus, a number of spores.

Throughout the world, the spread of microbes resistant to all known antimicrobials is in great danger. Therefore, it is extremely important to search for new chemical compounds with biocidal properties against a wide range of opportunistic and pathogenic microorganisms.

The objective of the invention is to obtain new compounds with a wide spectrum of antimicrobial activity with low toxicity to expand the raw material base and the range of antiseptic and disinfectants.

To solve this problem, copolymers of diallylaminophosphonium salts with sulfur dioxide of the general formula

where A = Cl ^- or BF ₄ ^- , soluble in methanol, dimethyl sulfoxide, dimethylformamide or when A = Cl ^- soluble in water, obtained by copolymerization of equimolar amounts of sulfur dioxide and diallylamino-phosphonium salt selected from tris (diethylamino) diallylamino-phosphonium chloride or diallylaminophosphonium tetrafluoroborate.

The copolymers of the general formula (1) exhibit antimicrobial activity.

Obtaining diallylaminophosphonium salts

Tris (diethylamino) diallylaminophosphonium chloride (EAAF-Cl) is prepared according to the following procedure.

Freshly distilled allyl chloride is added to tris (diethylamino) phosphazohydride with vigorous stirring and stirring of the reaction mixture is continued until the temperature rises spontaneously. Further, without interrupting stirring, sodium hydroxide in the form of a 50% aqueous solution is dosed into the reaction mixture, while the temperature is maintained no higher than 38 ° C. At the end of the dosage, the reaction mixture was stirred for another 30-45 minutes and then kept under stirring for 10-12 hours under gentle boiling. After cooling the reaction mixture, the middle layer was separated and extracted with EAAF-Cl with dichloromethane. The extract is evaporated on a rotary evaporator at a temperature of 100 ° C in steps: first in a vacuum of a water-jet pump, then in a vacuum of an oil pump (5-7 mm Hg).

To obtain tris (diethylamino) diallylaminophosphonium tetrafluoroborate (EAAF-BF ₄ ) use the following method.

EAAF-Cl is dissolved in distilled water and, with vigorous stirring, a 20% aqueous solution of sodium tetrafluoroborate is added dropwise. The reaction mixture is stirred for 15-20 minutes and incubated for 24 hours at room temperature. Next, the precipitate is filtered off, washed with distilled water and dried in vacuum at a temperature of 80 ° C to constant weight.

The purity of the obtained compounds was controlled by elemental analysis and ¹³ C NMR. The value of chemical shifts (δ, ppm) and the multiplicity of the NMR spectrum

¹³ C EAAF-Cl and EAAF-BF ₄ are shown in table 1.

The copolymerization of diallylaminophosphonium salts with sulfur dioxide

The copolymerization of diallylaminophosphonium salts with SO ₂ was carried out according to the known method [Vorobyeva A.I., Vasilieva E.V., Gaysina H.A., Puzin Yu.I., Leplyanin G.V. The copolymerization of N, N-dialkyl-N, N-diallylammonium halides with sulfur dioxide // High molecular weight compounds. - 1996, t. 38, No. 10. - S.1663-1667].

Sulfur dioxide was dried by passing through concentrated H ₂ SO ₄ and freshly calcined CaCl ₂ . As an initiator, dinitrile azoisobutyric acid (DAC) was used. The copolymerization was carried out at a temperature of 80 ° C for 7 hours. The polymers were purified by double reprecipitation from a solution in an appropriate solvent (methanol) to a precipitant (tetrahydrofuran) and dried in vacuum to constant weight at 50 ° C.

The composition of the copolymers of the general formula (1) was calculated according to the results of elemental analysis.

¹³ C NMR spectra of EAAF-Cl and EAAF-BF ₄ copolymers with SO ₂ are shown in Table 2.

NMR spectra were recorded on a Bruker-AM-300 spectrometer with an operating frequency of 75.5 MHz for carbon nuclei. The spectra were recorded in proton broadband isolation modes and in JMOD mode. As the solvent used DMSO-d ₆ and D ₂ O; the internal standard is tetramethylsilane (TMS) and 2,2-dimethyl-2-silapentane-5-sulfonic acid, respectively.

The analysis of the spectra indicates that EAAF-Cl and EAAF-BF ₄ copolymerize with SO ₂ with the participation of both double bonds via intramolecular cyclization to form pyrrolidine structures (table 2).

The possibility of obtaining diallylaminophosphonium salts and their copolymers with sulfur dioxide is confirmed by the following examples.

Example 1. EAAF-Cl was obtained according to the described method.

The starting materials were taken in the following amounts:

Tris (diethylamino) phosphazohydride - 392 g (1.5 mol)

allyl chloride - 600 g (7.5 mol)

50% aqueous sodium hydroxide solution - 255 g (6.4 mol)

Received 490 g of EAAF-Cl. The yield is 86.5% of the theoretical.

Found,%: C 56.82; H 11.06; N, 14.55. C ₁₈ H ₄₀ ClN ₄ P.

Calculated,%: C 57.07; H 10.57; N, 14.79.

Example 2. EAAF-BF _{4 was} obtained by the described method.

The starting materials were taken in the following amounts:

EAAF-Cl - 490 g (1.3 mol)

distilled water - 2.0 l

20% aqueous solution of sodium tetrafluoroborate - 960 g

Received 390 g of EAAF-BF ₄ . The yield is 70.1% of theoretical.

Found,%: C 49.72; H 10.03; N, 12.83. C ₁₈ H ₄₀ F ₄ N ₄ PB.

Calculated,%: C 50.23; H, 9.30; N 13.02.

Example 3. EAAF-Cl with SO _{2 was} obtained according to the specified method.

The starting materials were taken in the following amounts:

EAAF-Cl - 1.8925 g (50 mol%)

SO ₂ - 0.32 g (50 mol%)

DAK - 0.066375 g (3% wt.)

Yield 1.45 g (58.7%)

Example 4. EAAF-BF ₄ with SO _{2 was} obtained according to the specified method.

The starting materials were taken in the following amounts:

EAAF-BF ₄ - 2.149 g (50 mol%)

SO ₂ - 0.32 g (50 mol%)

DAK - 0.07407 g (3% wt.)

Yield 1.45 g (58.7%)

Study of toxicity and antimicrobial activity

The acute toxicity parameters of copolymers of diallylaminophosphonium salts with sulfur dioxide were determined on white outbred mice of both sexes weighing 18-22 g according to the known method [Prozorovsky VB, Prozorovskaya MP, Demchenko VM The express method of determining the effective dose and its errors // Pharmacology and Toxicology, - 1978, No. 4, p.497-502].

Toxicological tests showed that the claimed copolymers belong to the 4th class of toxicity [Hodge HE, Sterner LH Cit. by Spector. Spector WS (Ed.) Handbook of Toxicology. V.1, 1956] when administered through the stomach (LD ₅₀ >> 1000 mg / kg) and can be used for medical purposes.

The determination of antimicrobial activity was carried out by the method of double serial dilutions [Methods of experimental chemotherapy / Ed. G.N. Pershina. - M.: 1971. p.109] on the museum test cultures of Escherichia coli pcs. 25922; Staphylococcus aureus, pcs. 906; Staphylococcus saprophyticus, ATCC 15305; Micrococcus luteus, ATCC 4698; Bacillus antracoides, 1312; Bacillus subtilis, ATCC 6633; Bacillus proteus, pcs. "Flowers"; Candida albicans, 264/624; Salmonella spp. (State Research Institute of Standardization and Control of Medical Biological Preparations named after L.A. Tarasevich). An 18-hour agar culture (2.5 × 10 ⁵ microbial bodies in 1 ml of medium) was used in the experiments. For the study, solutions of the compounds in water or dimethyl sulfoxide were taken. The maximum of the tested concentrations is 1000.0 μg / ml. The tubes were incubated at 37 ° C, followed by seeding after 24 hours in tubes with beveled meat-peptone agar. The results were taken into account by the presence and nature of the growth of crops on a nutrient medium.

It was found that the inventive copolymers inhibit the growth of staphylococcus and micrococcus in the concentration range of 7.8-31.2 μg / ml The death of the yeast-like fungus Candida albicans was noted when the concentration of solimers was 31.2 μg / ml. The copolymers also cause the death of Escherichia coli at a concentration of 62.5 μg / ml; Salmonella spp. - at a concentration of 125.0 μg / ml; Bacillus subtilis - in the concentration range of 15.6-31.2 μg / ml; Bacillus proteus and spores of Bacillus antracoides - in the concentration range of 250.0-500.0 μg / ml.

The results of studies of antimicrobial activity are presented in table 3.

The proposed copolymers of diallylaminophosphonium salts with sulfur dioxide have the following advantages.

1. They have a high biocidal effect against a number of bacteria, as well as yeast-like fungi and spore-bearing crops.

2. They belong to the 4th class of toxicity when introduced into the stomach, which allows their use for medical purposes.

3. Do not cause corrosion of processed materials.

Table 1 Chemical shifts and multiplicity of the ¹³ NMR signals of EAAF-Cl and EAAF-BF ₄ Structure C ₁ , C _{1 '} C ₂ , C _{2 '} C ₃ , C _{3 '} C ₄ C ₅

49.27 132.54 119.58 40.58 13.09 t d t t to

49.65 134.08 119.65 41.02 13.73 t d t t to

table 2 ¹³ C NMR Spectra of EAAF-Cl and EAAF-BF ₄ copolymers with SO ₂ (DMSO-d ₆ , TMS) No. Compound Chem. shifts and multiplicity of signals, ppm * C ₁ C ₂ C ₃ C ₄ C ₅ one

c 52.60 36.71 51.98 41.27 13.94 t 53.34 38.77 54.82 40.37 13.94 t d t t to 2

c 50.57 34.90 50,42 39.40 12.93 t 51.04 36.94 52.14 39.35 12,43 t d t t to Note: t - triplet, d - doublet, k - quartet; * - stereoisomers.

Table 3 Antimicrobial activity of copolymers of EAAF-Cl with SO ₂ and EAAF-BF ₄ with SO ₂ No. p / p The name of the type (strain) of the microorganism Antimicrobial activity (MIC), μg / ml EAAF-Cl with SO ₂ EAAF-BF ₄ with SO ₂ one Escherichia coli. 25922 62.5 62.5 2 Staphylococcus aureus, pcs. 906 7.8 <31.2> 15.6 3 Micrococcus luteus, ATCC 4698 > 7.8 <15.6 <15.6> 7.8 four Staphylococcus saprophyticus, ATCC 15305 <31.2> 15.6 15.6 5 Candida albicans, 264/624 31,2 <62.5> 31.2 6 Bacillus antracoides, 131 <500.0> 250.0 500,0 7 Bacillus proteus, pcs. "Flowers" <1000.0> 500.0 > 250.0 <500.0 8 Bacillus subtilis, ATCC 6633 <15.6> 7.8 <62.5> 31.2 9 Salmonella spp. <125.0 125.0

Claims (2)

1. Copolymers of diallylaminophosphonium salts with sulfur dioxide of the General formula

where A = Cl ^- or BF ₄ ^- , soluble in methanol, dimethyl sulfoxide, dimethylformamide or when A = Cl ^- soluble in water, obtained by copolymerization of equimolar amounts of sulfur dioxide and diallylamino-phosphonium salt selected from tris (diethylamino) diallylamino-phosphonium chloride or diallylaminophosphonium tetrafluoroborate. 2. The copolymers according to claim 1, exhibiting antimicrobial activity.