UA32599U - Polyoxypropyleneglycol guanidine as antibacterial agent - Google Patents

Abstract

Polyoxypropyleneglycol guanidine of formula:,wherein: R - CHCH(CH),n=4-34, m=8-12,as antibacterial agent.

Description

Description of the invention

A useful model refers to derivatives of guanidine, its salts, complexes or addition products, 2 specifically to polyoxypropylene glycol anidine of the formula:

Drt- dn MN nn o zhh p

МNNСЯ in de K-СНХСН(СН») p-4-34, t-8-12. as a bactericidal substance and intended for use in the microbiological industry, as well as for water treatment, as a disinfectant and antiseptic. 75 The literature lists bactericidal substances that contain secondary and tertiary amino groups (1-43.

Alkoxymethylmethylammonium metasulfate, commercial name Alcamon DS, is a well-known bactericidal substance

IB) formulas:

ISPnaln OSNoM (SNU SNoSN")" (SNzZO, p-10-18

It is widely used as a disinfectant for the treatment of livestock farms, veterinary, industrial and medical equipment.

But it is characterized by insignificant bactericidal activity against conditionally pathogenic gram-positive and gram-negative bacteria. In addition, it is not produced in Ukraine.

The task of a useful model is the creation of polyoxypropylene glycolguanidine, which provides bactericidal activity against gram-positive and gram-negative bacteria. in)

The task is accomplished by obtaining polyoxypropylene glycol guanidine of the formula:

Ministry of Emergency Situations Ge») where K-СНСН(СН») Со) p-4-34, t-8-12.

The essence of a useful model is explained by the following examples: see

Linear aliphatic polyoxypropylene glycol guanidines were synthesized by the reaction of oligooxypropylene glycols with "U" terminal amino groups and guanidinium chloride. The reaction takes place in one stage. The reaction scheme and structural formula of the obtained polymers can be presented as follows:

МНН СИ « to shk r, also Kovsh and n- s i oi - МН 8 ---| nn'

We have the INN "with SHI" where В - СН-СН(СНа) p-4-34, t-8-12.

Example 1. The starting components for the synthesis were di-oligopropylene glycol with terminal amino groups D-230 and guanidinium chloride. Before synthesis, di-oligopropylene glycol and guanidinium chloride were dried in a vacuum at 80°C for 2 hours to remove moisture. The percentage content of amino groups was determined by the titrimetric method, it was 69.895. "se 50 The synthesis of this polymer was carried out in bulk at a molar ratio of components of 1:1 at a temperature of 1502 and under vacuum conditions, in which ammonia is removed from the reaction mixture. c Molecular weight was determined by gel permeation chromatography: Myp-2860, Mmzh-5291, Mp/Mt-1.85, t:12.

Control over the completion of the reaction was carried out by the method of 14-spectroscopy (disappearance of the band at 3450 cm 7 - valence vibrations of primary amino groups). The final product was reprecipitated from alcohol to hexane. The reaction product dissolves in water, alcohol, DMF, and is a viscous substance. The surface-active properties of the synthesized product were studied and the following values of the characteristics of aqueous solutions were obtained: ugdipy742.46mN/m; KKM-5,12. 10-2mol/l, K-2.2.10Nm2/Kmol. in Fig. The IR spectrum of polyoxypropylene glycolguanidine is shown, the absorption bands of groups are shown on the abscissa, and the intensity of the absorption bands on the ordinate.

Absorption bands at 3300 and 3100 cm are observed in the IR spectrum, which correspond to valence vibrations

MH groups of guanidine and oligooxypropylene glycol diamine, 2800-297Osm"! absorption bands correspond to valence vibrations of CH", CH, CH", 1110cm7- corresponds to C-O vibrations, 166Osm" to deformation vibrations of MH. 65 Example 2

The initial components for the synthesis were di-oligopropylene glycol with terminal amino groups D-400 and guanidinium chloride. Before synthesis, di-oligopropylene glycol and guanidinium chloride were dried in a vacuum at 80°C for 2 hours to remove moisture. The percentage content of amino groups was determined by the titrimetric method, it was 40.095.

The synthesis of this polymer was carried out in bulk at a 1:1 molar ratio of components at a temperature of 1502 and under vacuum conditions, in which ammonia released during the reaction is removed from the reaction mixture.

The molecular weight was determined by gel permeation chromatography: Myp-4100, Mmzh-7831, Mp/Mt-1.91, volume 10.

Control over the completion of the reaction was carried out by the method of IR spectroscopy (disappearance of the band at 3450 cm 7 - 70 valence vibrations of primary amino groups). The final product was reprecipitated from alcohol to hexane.

The reaction product dissolves in water, alcohol, DMF, and is a viscous substance. The surface-active properties of the synthesized product were studied and the following values of the characteristics of aqueous solutions were obtained: udip741.46mN/m; KKM-4, 12.102mol/l, K-42.10N.m2/Kmol

Example C

The starting components for the synthesis were di-oligopropylene glycol with terminal amino groups D-2000 and guanidinium chloride. Before synthesis, di-oligopropylene glycol and guanidinium chloride were dried in a vacuum at 80°C for 2 hours to remove moisture. The percentage content of amino groups was determined by the titrimetric method, it was 8.095.

The synthesis of this polymer was carried out in bulk at a molar ratio of components of 1:1 at a temperature of 1502C and under vacuum conditions, in which ammonia released during the reaction is removed from the reaction mixture.

The molecular weight was determined by gel permeation chromatography: Mp-16800, Mzh/-36120, Mp/Mt-2.15, Tv.

Control over the completion of the reaction was carried out by the method of IR spectroscopy (disappearance of the band at 3450 cm'"! -valence vibrations of primary amino groups). The final product was reprecipitated from alcohol into hexane.

The reaction product dissolves in water, alcohol, DMF, and is a viscous substance. The surface-active properties of the synthesized product were studied and the following values of the characteristics of aqueous solutions were obtained: uplip38.46mN/m; KKM-4, 12.10 Zmol/l, K-6, 2.103N.m"/Kmol

To study the bactericidal activity of the synthesized compounds against gram-positive and gram-negative bzo bacteria, strains of bacteria from the Czech (SCM), American (ATSS) and Ukrainian (UKM) collections of microorganisms and some isolates that are currently supported in the scientific and methodological ( 2) laboratories of the microbiological profile of the Department of Biology of the NaUKMA, in particular: EzsPegispia soii VE, Rzepdotopaz and Aegidipova SSM 1961; Zegtaya tagsezsepa SSM 1257; Epiegorasieg sioaseave; Kierzivya rpeshtopiae; Misgosossyv

Isheiv SSM 1697yp; vgarpuosossiz atsgeiz SSM 209; Sogupebasiegist attopiadepez ATSS 68717Й7-UKM As-732;. EM Soguperasiegisht maparbie ATSS 15753И0-UKM As-717; VPodosossiv eguygoroiv UKM As-741ip and Vasiyiv so z,ibiyv SSM 104 (Type - strain, typical for the species). All bacteria were cultured on meat-peptone agar (MPA,

Oiso, USA), endo diagnostic and differential medium (Rica, USA) was used for cultivation of E. coli.

Bactericidal activity and minimum inhibitory concentration (MIC) of guanidine-containing compounds against gram-positive and gram-negative bacteria were determined by the method of dilutions in meat-peptone agar in -o c modification of replicas (7). The suspension of daily test cultures was prepared in sterile Masi's physiological solution. и Standardized according to the optical standard of turbidity, a suspension of bacterial cells (concentration 1.109 cells/ml) in a volume of 0.2 ml was introduced into the wells of the replicator and sown in parallel in three Petri dishes on the surface of the MPA nutrient medium containing different concentrations (ppt) of polyoxypropylene glycol guanidines (examples 1, 2 and 3). The results were calculated 24 and 48 hours after the cultivation of bacteria at 3290. As a control, the medium without the addition of polyoxypropylene glycol guanidines was used. The minimum inhibitory concentration was considered to be the smallest concentration of the compound in the medium at which the growth of microorganisms was absent.

Ge) The results of determining the bactericidal effect of the synthesized compounds prove that the synthesized "c 20 polyoxypropylene glycol guanidines (examples 1, 2 and 3) show an inhibitory effect on gram-positive and gram-negative bacteria. The compound given as example 1 (the) (table 1) is characterized by the highest bactericidal activity. and regarding various representatives of gram-positive and gram-negative bacteria c)

ATSSbVT1 TYPE (-UKM As-732)

Sogupebrasiegiit magiabiie with -4 -4 -4 -4 -4 -4 -4 for kesvoeyuetya 707 b5 KPodosossiv eguipgoroiiv tb --4 A A A A A A A zani 5

UKMAS-741 Type

Захуснетнсиннссмой | ee) not 20085 nt

Vaznezinssmusue 00000032 o00oveaeensoyut 000 ння нння нет нн fнн| yan ie tyeeetv sm ee nnenne nn ne ee 000000 nennie vnie Ge ee en ye ve ten yayu 70 1961 century: NI Z la v v EN number of repetitions. The intensity of culture growth was determined according to a 4-point system, respectively, marked as: "-" - no growth of microorganisms, "-" - growth of single colonies, "k-" - weak continuous growth, "nc" - continuous growth, "zhk" " - continuous abundant growth, 75 similar to the growth in the control.

Determination of the minimum inhibitory concentration for various test cultures made it possible to find out that with an increase in the molecular weight of polyoxypropylene glycol guanidines, their bactericidal activity decreases slightly (table 2). The results of a comparative assessment of the sensitivity of test cultures - representatives of different genera of gram-positive and gram-negative bacteria are presented in Table 2. for different representatives of gram-negative and gram-positive bacteria and C

F zo

F

Vvrpuyusossiv am SSMOYUU 11111110 yu cm zv s « 8 : microorganisms, the table shows summarized data, after sowing cultures on 3 Petri dishes. with

As a result of the study of the bactericidal activity of polyoxypropylene glycol guanidines against different groups of bacteria, it was established that they exhibit the maximum bactericidal effect against gram-positive bacteria. se At a concentration of 25O0ppm, none of the investigated test cultures - representatives of various genera of gram-positive bacteria - grew. The strains Misgosossiz ISheiv SCM 169 Type and Soguperasiegicht attopiadepez ATS 68717y7-UKM As-732 were the most sensitive to the mentioned compounds. The MIC for these test cultures for all tested compounds (examples 1, 2 and 3) did not exceed 25ppm (table 2). Somewhat higher IPC was for Ziarpuiosossiv asheyv SCM 209; Soguperasiegicht magyarie ATSS 15753 and 1"-UKM As-717; Vpodosossiv eguygoroiiz UKM As-741 Type o and Vasiiyz zibiyv SSM 104, which, respectively, was 250 ppt. (Te) Representatives of gram-negative bacteria turned out to be more resistant to polyoxypropylene glycolguanidines. It should be noted that polyoxypropylene glycolguanidines are practically the same affected all examined gram-negative test cultures. In contrast to gram-positive bacteria, the MIC for gram-negative cultures was higher than 250ppm. Thus, as a result of the research, it was established that a concentration of 1000ppm of polyoxypropylene glycol guanidines in the medium guarantees 10095 death of all tested test cultures. c According to the literature, the well-known bactericidal substance Alkamon DS is widely used for disinfection of equipment in medical institutions and the food industry, disinfection of hands, remediation of the oropharynx, etc.

This compound is characterized by a selective antimicrobial effect and has a more active effect on gram-positive bacteria, in particular, Ziarpuiosossiv atsgeiz |5), than on gram-negative bacteria, namely - Rzepdotopaz aegodiposa 6). A comparative analysis of the BET of polyoxypropylene glycolguanidines with the prototype (alkamon DS) revealed that they are characterized by bactericidal activity higher than that of the prototype, both against gram-positive and gram-negative bacteria (table 3). Thus, the MIC of alkamone DS was 21 and 4 times higher in gram-positive bacteria, respectively, for examples 1 and 2, and 1.6 times - for gram-negative bacteria. b5

Table C

Minimum inhibitory concentrations of polyoxypropylene glycol guanidines against gram-negative and gram-positive bacteria

AND

(Ziarpuyusossiv howling SSM0Ю 011101 5О lev

The advantage of the synthesized polyoxypropylene glycol guanidines is a higher bactericidal activity against gram-negative and gram-positive bacteria, compared to such a well-known bactericidal substance as alkamon dos

Sources: 1. GDR patent Mo215708, Method of obtaining epoxy amine adducts, January 21, 1981. 2. Application of the Federal Republic of Germany Mo3736995, Synthetic resin containing nitrogen groups and its purpose, 11.05.89.

Z. M. Oeziaiv, Yu. Adevz, 9U. Zatse. Roiut. Vii., 2000, 44, 401. 4. Masgotoies. Maieg. Epd., 2001, 286, Mog, pp. 63-87. 5. Radchenko O.S., Stepura L.G., Furtat I.M., Mikhalskyi L.O., Kramarev SO. Prospects of using quaternary ammonium compounds for sanitation of mucous membranes and disinfection of wounds /L/infectious diseases. - 2002. - Mo1. - P.59-63 - prototype. 6. Stepura L.H., Radchenko O.S., Furtat I.M., Mikhalskyi L.O. Bactericidal properties of cationic and other surface-active substances // Agroecological Journal. - 2002. - Mo4. - P.60-63. 7. Vasilevskaya I.A., Sergeychuk M.G., Zgonnyk V.V., Furtat I.M., Mikhalsky L.A. Improvement of the method of bacteriological control in the production of lysine // Mikrobiol.zhurn. - 1996. - 58, Mob. - P.66-76.

Claims (1)

The formula of the invention Z Polyoxypropyleneglycolguanidine of the formula: FE МН МН МН re thoi (22) МН МНС и юю де: К - SNASN(СНУ»), с Зо п-4-34, т-8-12, (ze) as a bactericidal substance . - with ;" with co 1 (Se) (32) p. 60 b5