RU2645137C1 - Bioicide sorbent based on chlormethylated styrene copolymers - Google Patents

Abstract

FIELD: chemistry; processing and recycling of wastes.

SUBSTANCE: invention relates to sorbtion materials for purification and disinfection of aqueous and gaseous media. Biologically active sorbent comprises a porous sorption material substrate from a chloromethylated styrene copolymer and a divinyl-containing linking agent. As the biologically active component, heterocyclic skeletal compound 1,3,6,8-tetraazatricyclo[4.4.1.1^3,8]dodecane (teotropine) is applied to the substrate. Teotropin is applied to the substrate by the amination reaction of chloromethyl groups of the copolymer with teotropin to form covalent bonds in the presence of organic solvents selected from the classes of higher alcohols and glycols.

EFFECT: invention provides sorbent with bactericidal properties of a wide profile and ability to increase the plasma clotting time.

3 cl, 1 tbl, 4 ex

Description

The invention relates to the field of development of sorption materials for cleaning and disinfecting water and gas environments in order to ensure safe life, as well as for possible use in medicine for decontamination of biological body fluids: blood, plasma, cerebrospinal fluid, lymph and pharmacology for the disinfection of technological and injection solutions.

The need for such developments is associated with the need to combat pathogens of dangerous infectious diseases and the possibility of their use for terrorist purposes. First of all, they include causative agents of anthrax, tularemia, plague, botulism, smallpox, hemorrhagic fevers, mutating strains of influenza, glanders, typhus, as well as a group of diseases of agricultural and experimental animals. At the same time, it is very important to prepare water and food liquids for humans and agricultural and experimental animals, as well as the sorption disinfection of water, which is necessary for sanitary and hygienic purposes in solving problems of the Belarusian Railways and emergency situations.

One of the oldest and most reliable methods suitable for the preparation of water for humans and animals is its passage and filtration through a column with a sorbent having bactericidal, sporocidal, virucidal or fungicidal properties. At the same time, microorganisms, viruses, spores in water come into contact with a bactericide that is contained on the surface of the stationary phase of the sorbent.

For these purposes, bactericidal sorbents are known, which are hard inorganic bactericides and compositions based on them, for example, iodine, sulfur and silver-containing components sorbed on ion-exchange resins or carbon sorbents.

Bactericidal sorbent used in filters for disinfecting water from a water supply system and freshwater sources is known from the prior art. Carbon-containing material is used as a sorbent, and iodine-containing and silver-containing components are used as a bactericidal additive [patent RU 2221641 C2, publ. January 20, 2004].

Also known from the prior art is the use of iodine-containing anion-exchange resin, granular activated carbon, cation-exchange resin and anion-exchange resin with a silver-containing sorbent for disinfecting water in a layer of granules [patent RU 2172720 C1, publ. 08/27/2001].

The prior art also describes a composition intended primarily for the disinfection of natural and waste water, drinking water from open fresh water bodies (rivers, lakes), underground sources (artesian water intake) and drinking water supply. The composition contains as a biocidal additive a silver-containing preparation, including a solution of iodine or hydrogen peroxide, or potassium permanganate, and as a sorbent - glauconite or activated carbon [patent RU 2191163 C1, publ. 10/20/2002].

Known iodine and silver-containing ion exchangers have a number of disadvantages, the main of which are significant excretion of strong oxidizing agents of iodine, triiodide ion and highly toxic silver ions into plasma or blood. In addition, silver and iodine-containing substances are clearly limited in the range of their application only to bactericidal properties and, to a much lesser extent, to virucidal properties, and their effect does not extend to biologically more dangerous fungal and spore forms.

Most inorganic bactericides are distinguished by relatively high toxicity and the inability to exclude interaction with blood or plasma proteins, which limits the use of these sorbents for the purification of biological body fluids on sorption columns by autoplasmapheresis.

Synthetic bactericides are generally milder and have a relatively low species toxicity to mammals.

As the closest analogue (prototype) of the present invention can be considered the document [patent RU 2312705 C1, publ. December 20, 2007], which discloses a highly effective biocidal sorbent obtained by fixing the currently widely spread theotropin bactericide with ionic bonds on cation exchange resins of various nature (sulfo-, phosphoric, carboxylic) with the formation of cationite-theotropin ionic bonds.

The use of theotropin made it possible to expand the spectrum of action of the polymer bactericide, primarily to the causative agents of anthrax, tularemia, plague, botulism, smallpox, hemorrhagic fevers, glanders, typhoid fever and Venezuelan encephalitis. The prototype shows that the use of theotropin allows you to achieve the concentrations necessary to suppress these infections. In this case, theotropin itself belongs to class 4 and shows a toxic concentration of about 500 mg / kg of live weight. Theotropin is isolated from the solid phase upon contact with aqueous media by hydrolysis with the cleavage of theotropin sorbate, and sufficient amounts of the bactericide appear in the aqueous solution, which exerts its effect.

The main problem limiting the use of polymeric bactericides of a known type for the purification of native biological body fluids and food fluids is the intensive leaching of toxic chemically active bactericides into the treated medium. So silver, according to the degree of harmful effects on higher organisms, belongs to the 2nd hazard class, highly hazardous substances - the average lethal dose when administered to the stomach 15-150 mg / kg. According to various sanitary standards, the maximum concentration of silver in drinking water is from 0.05 to 0.001 mg / l, which is comparable to cumulative supertoxicants such as mercury and lead (maximum concentration of 0.001-0.03 mg / l).

The transition in the prototype to synthetic organic bactericides with lower toxicity allowed the creation of more acceptable systems for disinfecting liquid media. So theotropin belongs to hazard class 4 and shows significantly less toxicity compared to silver - the maximum permissible dose of about 500 mg / kg live weight.

For such systems with ionic bonds "bactericide - cation exchanger" is characterized by low retention capacity and rapid hydrolysis of weak ionic bonds. As a result, upon contact with aqueous media, a rapid transition of the bactericide to the disinfected medium is observed. The presence of metal ions in biological fluids of the body, for example, calcium and magnesium, or high molecular weight oligoproteins, which effectively displace low molecular weight theotropin from sulfocathionite, leads to a high excessive concentration of theotropin in a disinfected (decontaminated) environment, such as blood plasma, and increases the risk of toxic effects on the body.

In the process of disinfecting the medium by passing it through a layer in the prepared mineralized water and body fluids, an excessive concentration of theotropin is created in excess of the necessary for effective bactericidal activity and complete suppression of microorganisms. As a result of fixation of theotropin by weak ionic bonds and rapid displacement of theotropin, the concentration of toxic bactericide in the medium being cleaned increases and the time for maintaining bactericidal activity or the specific volume of the disinfected medium per layer volume decreases.

The technical result of the present invention is to create a sorbent with bactericidal properties of a wide profile, including pathogenic and superpathogenic infections, and characterized by contact with plasma with the ability to increase plasma clotting time. Moreover, such sorbents can provide a bactericidal effect both upon contact with the sorbent solution, and during dynamic disinfection in the duct with various forms of filter elements, i.e. its use in the form of granules, foam (sorbent with developed porosity), composites, napkins, fibers or tow, as well as film and non-woven material.

In addition, the process of plasma disinfection on the proposed sorbents is accompanied by the ability to increase the coagulation time of the plasma and its approximation in terms of MHO (international normalized ratio) to a special plasma for the treatment of thromboembolism.

The indicated technical result is achieved by a biologically active sorbent with biocidal properties, including a substrate of porous sorption material, onto which a heterocyclic frame-type compound of 1,3,6,8-tetraazatricyclo [4.4.1.1 ^3,8 ] dodecane is applied ( theotropin), and characterized in that the substrate of the porous sorption material is a chloromethylated copolymer of styrene and a divinyl-containing crosslinking agent, and the application of theotropin on the substrate carried out by amination of the chloromethyl groups of the copolymer with theotropin to form covalent bonds, the amination reaction is carried out in organic solvents selected from the classes of higher alcohols and glycols, in the temperature range 40-70 ° C for 2-8 hours.

In addition, the biologically active sorbent proposed in the present invention is characterized by the ability to increase the coagulation time of plasma.

In a preferred embodiment, the divinyl-containing crosslinking agent is divinylbenzene, and its amount is 2-30 mass. %, and the content of active chlorine in chloromethyl groups is in the range of 6-18%.

In another preferred embodiment of the invention, the substrate of porous sorption material is used in the form of granules, porous plastic (sorbent with a developed surface), composites, fibers, tow or non-woven material.

Since the type of substance applied determines the spectrum of suppressed microorganisms, the choice of theotropin can significantly expand the spectrum of bactericidal action, including the suppression of pathogens of a group of human infectious diseases, as well as agricultural and experimental animals: anthrax, tularemia, plague, botulism, smallpox, hemorrhagic fevers, mutating strains flu, glanders, typhus and others, to which the high effectiveness of theotropin-based drugs and compositions is known.

The proposed chloromethylated copolymer - theotropin system provides higher bactericidal activity in disinfected media, which can be seen from the values of the average thickness of the zone of complete suppression (ZPP) for standard strains according to the results of 21 and 42 hours of incubation.

The present invention provides sorbents containing active fragments of theotropin obtained on the widespread chloromethylated copolymers, which are the basis for anion exchange resins, by bonding theotropin on a polymer substrate with stronger covalent bonds. The binding of theotropin to chloromethyl groups leads to the formation of very weak quaternary ammonium bases, which undergo cleavage or hydrolysis, giving theotropin hydrochloride and methylol groups instead of chloromethyl.

The claimed invention proposes to fix theotropin in the polymer structure due to weak hydrolyzable forms of quaternary ammonium bases. There is another possibility of disinfection, which is associated with the formation of clathrate structures with water, which have a pronounced bactericidal effect. Some of these components were known as substances that damage the cell wall of highly dangerous microorganisms.

The main advantage of the proposed method is the absence of large concentrations of the bactericide in the passing water. That is what allows you to use this method for purification of body fluids.

Another advantage is the ability to create based on the proposed sorbents and their additional modifications. Styrene copolymers containing a chemically active alkyl substituted benzene ring make it possible to easily carry out chloromethylation reactions under fairly mild conditions and to obtain high conversions with the introduction of chloromethyl groups. At the same time, it becomes possible to use them not only in the form of a bulk filter, but also in the form of a poroplast (sorbent with a developed surface), various composites containing alkyl substituted aromatic rings, nonwoven materials, as well as fibers or tows that can be used for packing narrow columns and creating a stable sorption layer. This allows the development of highly efficient sorption filters for the disinfection of liquid and gas media. For the latter, non-woven material is most suitable, which has up to 95% voids and creates small resistance to the movement of gas flows. It is possible to use it in any devices supporting the vital activity of humans or farm animals, as well as in various vivariums for keeping experimental animals.

The grafting of polystyrene easily takes place on various surfaces, and the styrene copolymer undergoes a further effective chloromethylation reaction to form active chloromethylated derivatives. This facilitates the subsequent reactions of addition or fixation of theotropin.

Examples 1-4 demonstrate the implementation of the present invention. The examples are illustrative and in no way limit the scope of the claims.

Example 1

In a 500 ml three-necked reactor equipped with a low-speed stirrer (speed 2 rpm), a contact thermometer and a reflux condenser, 80 ml (≈35 g) of air-dried chloromethylated macroporous copolymer of styrene and divinylbenzene (DVB) (30 wt% DVB) are placed , the blowing agent is isooctane 115 wt.%, chlorine content - 18.8 wt.%), make 320 ml of theotropin solution with a concentration of 117.9 g / l (0.702 mol / l) in a mixture of solvents (ethanol and propylene glycol in a ratio of 25: 75 vol.%) And incubated with continuous stirring for 4 hours at tamper round 40 ° C.

After that, the contents of the reactor are separated into solid and liquid phases on a funnel, the solid phase is washed successively in a Buchner funnel with distilled water (until theotropin is absent in the outlet) and 48 ml of acetone (60% of the volume of the initial chloromethylated copolymer) and dried in air. The content of theotropin in the solid phase is determined by the gravimetric method and in parallel with potentiometric titration to reduce theotropin concentrations in the liquid phase. The experimental gain was 0.51 g / g of dry chloromethylated copolymer, the amount of fixed theotropin was 3.03 mmol / g of dry chloromethylated copolymer. The liquid phase after theotropin addition is possible in recycling to obtain the next portion of the polymer bactericide.

Example 2

Carried out in accordance with example 1. In a three-necked reactor load 80 ml (≈40 g) of air-dry chloromethylated gel copolymer of styrene and divinylbenzene (8 wt.% DVB, chlorine content - 12.2 wt.%), Then add 320 ml of solution theotropin with a concentration of 82.5 g / l (0.491 mol / l) in a mixture of solvents (ethanol and propylene glycol in a ratio of 25:75 vol.%) and incubated with continuous stirring for 4 hours at a temperature of 50 ° C.

After that, the contents of the flask are separated into solid and liquid phases on a funnel, the solid phase is washed successively in a Buchner funnel with distilled water (until theotropin is absent at the outlet) and 48 ml of acetone (60% of the initial cation exchange resin) and dried in air. The content of theotropin in the solid phase is determined by the gravimetric method and in parallel with potentiometric titration to reduce theotropin concentrations in the liquid phase. The experimental gain was 0.40 g / g of dry chloromethylated copolymer, the amount of fixed theotropin was 2.1 mmol / g of dry copolymer.

Example 3

Carried out in accordance with example 2. In a three-necked reactor load 20 g of air-dry fiber coated with fiber and then with chloromethylated styrene polymer (chlorine content - 6.3 wt.%), Then make 320 ml of theotropin solution with an initial concentration of 49.5 g / l (0.295 mol / l) in a mixture of solvents (ethanol and propylene glycol in a ratio of 25:75 vol.%) and incubated with continuous stirring for 4 hours at a temperature of 20 ° C.

After that, the contents of the flask are separated into solid and liquid phases on a funnel, the solid phase is washed successively in a Buchner funnel with distilled water (until theotropin is absent in the outlet) and 48 ml of acetone (60% of the volume of the initial fiber) and dried in air. The content of theotropin in the solid phase is determined by the gravimetric method and in parallel by reducing the concentration by potentiometric titration. The experimental gain was 0.22 g / g dry cation exchanger, the amount of fixed theotropin 1.23 mmol / g dry chloromethylated fiber.

Example 4

As a control sample for this example, the starting macroporous styrene copolymer with 20 wt. % divinylbenzene with a blowing agent - isooctane 115 wt. % and chlorine content - 18.8 mass. % without applying theotropin.

The achieved level and technical result of the present invention is confirmed by studies on standard strains, as well as analyzes of human plasma, performed in certified specialized laboratories.

The study of bactericidal activity was carried out according to the guest method for the option of a hole on a lawn (meat-peptone agar) in standard Petri dishes with a diameter of 90 mm, a hole diameter of 8 mm with fixation of partial (ZPCh) and complete (ZPP) suppression zones with averaging over eight sectors with a step of 45 °. The experimental density on a lawn 4 mm thick: E. coli K-12, S. aureus 290-P - 10 ⁸ CFU / ml, B. anthracis - 10 ⁶ CFU / ml 55-VNIIVViM as the turbidity standard, the commercial McFarland standard was used.

Table 1 shows the comparative values of the average radius of the zones of complete suppression (ZPP) of standard strains for two contact times of 21 and 42 hours in E coli (Escherichia coli) E.c. 21, E.c. 42 and Staphylococcus aureus (lat. Staphylococcus aureus) S.a. 21, S.a. 42. Each series was performed according to the guest procedure for four weighed portions of the tested polymer bactericides (5, 10, 20, 40 mg) on one Petri dish.

As can be seen from the table, the developed materials by the values of the zones of complete suppression have a fairly good bactericidal effect, 1.5-2 times higher than those for the prototype.

Moreover, the values of the RFP for 42 hours of temperature control are equal to or greater than for 21 hours, which indicates the duration and stability of the achieved effect. Values of RFP for S.a. 1.5-2 times higher than for E.s., i.e. the effect on saprophytes is less than for pathogens.

In order to use sorbents as hemo- or plasmosorbents, hemo- and plasmocompatibility requirements must be additionally presented to them accordingly. For the samples in examples 1-4, a number of indicators were determined that determine the coagulation ability of plasma before and after contact according to generally accepted clinical parameters: clotting time, content (in%) of prothrombin and antithrombin III, as well as the integral indicators of MHO (international normalized ratio).

All proposed compositions of polymer bactericides for plasma analysis after 3-hour contact did not show negative changes leading to accelerated coagulation or an increase in prothrombin or the prothrombin / antithrombin III ratio as indicators of coagulation, which indicates their good compatibility with plasma. The resulting sorbents, as well as the initial chloromethylated copolymers, mainly absorb prothrombin and complexes of prothrombin with antithrombin III from the plasma. In this case, the coagulation time is significantly increased, up to the absence of coagulation within 40-180 s. Thus, the proposed sorbents in contact with plasma can achieve the effect of hypocoagulation, that is, increase the coagulation time until it disappears and do not give a negative effect of hypercoagulation.

Claims (4)

1. A biologically active sorbent with biocidal properties, including a substrate of porous sorption material, onto which a heterocyclic frame-type compound 1,3,6,8-tetraazatricyclo [4.4.1.1 ^3,8 ] dodecane (theotropin) is applied as a biologically active component characterized in that the substrate of porous sorption material is a chloromethylated copolymer of styrene and a divinyl-containing crosslinking agent, while theotropin is applied to the substrate by the chlorination amination reaction alkyl groups of the copolymer with theotropin with the formation of covalent bonds, and the amination reaction was carried out in organic solvents selected from the classes of higher alcohols and glycols, in the temperature range 40-70 ° C for 2-8 hours. 2. The biologically active sorbent according to claim 1, characterized in that it is characterized by the ability to increase plasma clotting time. 3. The biologically active sorbent according to claim 1 or 2, characterized in that the divinyl-containing crosslinking agent is divinylbenzene, and its amount is 2-30 wt.%. 4. The biologically active sorbent according to claim 1 or 2, characterized in that the substrate of porous sorption material is made in the form of granules, foam, composites, fibers, tow or non-woven material.