RU2319488C1 - Entherosorbent - Google Patents

Abstract

FIELD: natural sorbents.

SUBSTANCE: disclosed is entherosorbent of wide action spectrum based on opal-crystoballite rock. Sorbent is useful in combined radiation and thermal lesion in human and animals, alcoholic and drug intoxication, as antibacterial agent.

EFFECT: improved entherosorbent.

2 cl, 12 ex

Description

The invention relates to the field of development and use of natural sorbents, in particular, the claimed sorbent can be used in medicine and veterinary medicine to relieve bacterial and tissue toxemia after nuclear and thermal damage to humans and animals, for the treatment of complications arising from combined radiation therapy of cancer patients, in the treatment burn disease and infectious diseases with a predominant lesion of the gastrointestinal tract, i.e. with infections accompanied by massive destruction of tissues. In addition, the claimed sorbent can be used to relieve or reduce alcohol or drug intoxication.

Currently, antibiotics of a wide spectrum of action are used to treat infections arising from massive radiation-thermal damage (RTP) in people. A combination of antibiotics and the elimination of developing tissue toxemia using enterosorbents is known (RU 2161486, 10.01.2001).

However, the use of several antibiotics after radiation and RTP does not eliminate the phenomena of tissue and metabolic toxemia caused by damage and decay of cells of radiosensitive tissues: lymphoid and hematopoietic system. Under these conditions, the body remains unprotected from the action of bacterial toxins and decay products of its own cells from radioactively damaged organs. Irradiation also causes a violation of the proliferation of cells of the hematopoietic system and intestinal epithelium; therefore, elimination of toxemia and restoration of cell proliferation after RTP is of primary importance for increasing animal survival and reducing or completely preventing toxic reactions in patients during radiation therapy. Currently known sorbents do not sufficiently eliminate bacterial and tissue toxemia and related metabolic disorders.

The use of tripoli is known for the Pervomaiskoye field of the Alatyr district of the Chuvash Republic and the Shadrinsk field of the Kurgan region. for enterosorption in the treatment of acute poisoning. Tripoli with a particle size of not more than 0.8 mm is introduced into the stomach through a probe as a sorbent. The amount of 20-50 g per 150-200 ml of water. The procedure is carried out 1-2 times with an interval of 2-5 hours (RU 2188645, 09/10/2002). The method improves the sorption of toxins and allows to shorten the patient's recovery from a coma.

However, there is no data on the effect of tripoli of these deposits on bacterial and tissue toxemia.

Known enterosorbent, which is a suspension of glauconite in an aqueous solution of agar, used to treat calves with dyspepsia (RU 2188652, 09/10/2002).

Known sorbent for the treatment of infected wounds made of zeolite powder in silver form (RU 2245151, 01.27.2005).

Enterosorbents based on zeolite-containing tuffs are known for the treatment of renal failure (RU 2097046, 11.27.1997), for the treatment of peptic ulcer (RU 2143909, 01.10.2000), as an anti-allergic agent (RU 2082403, 06.27.1997), for the treatment of alcohol intoxication (RU 2145216, 02.10.2000).

Closest to the proposed is enterosorbent-shivyrtuin, made from silica-containing raw materials, containing a total of 60-80% clinoptilolite and montmorillonite, as well as impurities of quartz and biotite. Enterosorbent exhibits sorption activity against microorganisms, selectivity for certain radionuclides, weakens the diffusion of toxins, stimulates the migration of leukocytes into the wound (RU 2122868, 10.12.1998).

However, the effect of this sorbent on metabolic and tissue toxemia is unknown.

The problem solved by the present invention is, on the one hand, to expand the range of enterosorbents, and on the other hand, to obtain enterosorbent with a wide spectrum of action on both metabolic and bacterial and tissue toxemia.

The problem is solved by the proposed enterosorbent based on silica-containing mineral raw materials containing sedimentary opal-cristobalite rock containing not more than 15 wt.% Clay minerals of the montmorillonite group and not more than 10 wt.% Minerals of the mica group.

Preferably, the enterosorbent contains opal-cristobalite rock of the Zikeevsky deposit of the Kaluga region.

In a particular case, enterosorbent is used for combined radiation-thermal damage.

In a particular case, enterosorbent is used for alcohol intoxication.

In a particular case, enterosorbent is used for drug intoxication.

The technical result of this invention is to reduce all types of intoxication in the body and to expand the range of enterosorbents.

Opal-cristobalite rocks are the main component of finely porous sedimentary rocks (flasks, tripoli) with the remains of globularia. Opal is a solid hydrogel of silicon dioxide, and cristobalite is the product of crystallization of opal. Such rocks usually also contain clay group minerals, mica, hydromica (up to 30% in total), impurities of quartz, calcite, and possibly feldspars. The adsorption properties of opal-cristobalite rocks are known, for example, from RU 2189650, September 20, 2002, RU 2219138, December 20, 2003. Opal-cristobalite rock is also used as a normalizing additive in the feed composition for farm animals and poultry (RU 2084178, 07.20.1997, RU 2077320, 04.20.1997, RU 2134042, 08.10.1999) and as a source of assimilable silicon for living organisms ( RU 2199883, 03/10/2003).

However, published sources lack information on the effect of opal cristobalites on metabolic and tissue toxemia, as well as information on the possibility of using rocks containing clay and micaceous minerals for enterosorption.

The applicant found that the content of clays of the montmorillonite group in the enterosorbent should not exceed 15 wt.%, Since the minerals of this group have a high swelling in liquids, and when their content is above 15%, difficulties arise in enterosorption through a probe. The mineral content of the mica group should be limited to 10 wt.%, Since with a standard degree of grinding of the raw material (0.1-1 mm), the edges of the mica plates are sharp enough, which can cause various problems in the body.

The applicant has established that enterosorbent based on opal-cristobalites of the Zikeevskoye deposit of the Kaluga region showed the greatest efficiency.

The content of the minerals that make up the rock was monitored using a DRON X-ray diffractometer with a copper emitter and a nickel filter. The quantitative content of clay and micaceous minerals was determined by the method of “decomposition of the multiplet”.

Below are the results of experiments with the declared enterosorbent (see legend - HCM), in which the content of montmorillonite was 8-10% and biotite mica 6-8%.

Studies were conducted in experiments in vitro binding of bacteria E. Coli; You. Subtilis, proteins and peptides - medium-weight molecules (MSM) in serum, vitamin B ₁₂ with the proposed enterosorbent-GKM and the comparison drug "Smecta" (France). In in vivo experiments in animals with RTP, the content of E. Coli in the small and large intestines, which is the organisms most seeded with microbes, was studied, as well as the content of peptides (MSM) and toxic oligopeptides (TOP) in rat blood serum 48 hours after RTP. The proposed enterosorbent (GCM) and the Smecta comparison drug in doses of 200 mg / kg in a volume of 0.5 ml of water are administered to rats 2 times during the first two days after exposure (immediately and 24 hours after RTP).

Example No. 1

In in vitro experiments, the sorbing properties of HCM and Smecta were evaluated by the sedimentation rate of gram-negative (Escherichia coli) and gram-positive (Bacillus subtilis) bacteria in liquid buffer in the presence or absence of these sorbents.

The choice of the type of bacteria is due to the fact that they are classical representatives of gram-positive and gram-negative microorganisms, representing two groups of prokaryotes and characterized by fundamental differences in the construction of the cell wall.

Preliminarily, GCM or Smecta, 500 mg each, was placed in glass tubes with stoppers and sterilized for 30 min at 150 ° C. After that, 5 ml of sterile buffer with microorganisms was poured into each tube.

Microorganisms were incubated for 18 h at 37 ° C in a nutrient broth (1% peptone, 1% yeast extract, and 0.9% NaCl). The cells at a concentration of 10 ⁸ cells / ml, centrifuged on an OPN-8UHL 4.2 centrifuge 4.2 No. 5117, for 10 minutes at 5000 rpm were resuspended in isotonic 0.01 mol / L phosphate buffer pH 7.0 containing a suspension of clay particles or Smecta . After this, the suspensions were mixed by shaking on a mixer-mixer.

The intensity of the sorption process was studied at 37 ° C, plating the cells, after various time intervals after incubation of the suspension of cells and these materials, on solid nutrient agar. Sowing of cell suspensions and estimation of the number of viable microorganisms was carried out using the drip method of microorganisms.

Example No. 2

In vitro experiments established that the concentration of E. Coli B / r cells in suspension without sorbents for 1 hour in the entire volume remained virtually unchanged, equal to 1.09 · 10 ⁸ cells / ml at the beginning of the study and 1 at the end 15 · 10 ⁸ cells / ml. The addition of HCM to the suspension led to the fact that in 1 hour the concentration of gram-negative bacterial cells decreased from 1.2 · 10 ⁸ to 3.6 · 10 ⁶ cells / ml. The sorbent Smecta studied as a comparison drug lowered the cell concentration from 1.2 · 10 ⁸ to 2.2 · 10 ⁶ cells / ml.

Example No. 3

The experiments with Bacillus subtilis revealed a similar pattern: in the buffer, the cell concentration did not change within one hour, the addition of Smecta reduced the concentration of bacteria by 39 times, and GKM - by 10.4 times. The initial concentration of cells in the buffer was 2.7 × 10 ^{8 -} 2.9 × 10 ⁸ cells / ml, an hour after the addition of Smecta, 6.4 × 10 ⁶ cells / ml remained in the supernatant, the introduction of HCM reduced the cell concentration to 2.8 × 10 ⁷ cells / ml.

Example No. 4

To determine the binding of rat blood serum proteins, sorbents were incubated with 1 ml of serum and 100 mg of Smecta sorbent or HCM in a water bath for 1 hour at 37 ° C. Then, using Bradford's reagent (Scopes R. // Methods of protein purification. - World. - 1985. S.342) determined the concentration of proteins in serum. In the control (without sorbents), the protein concentration was 65 g / L. After incubation with Smecta - 61 g / l, after incubation with HCM - 62.5 g / l.

Example No. 5

To determine the binding of peptides (MSM), 1 ml of serum was incubated for 1 hour with 100 mg of sorbent: Smecta or HCM at 37 ° C in a water bath. Then, according to the method of N.I. Gabrielyan (Laboratory work. - 1984. - No. 3. - P.138-140), the number of MSM was obtained and determined. In the control serum (without sorbents), the number of MSM was 0.37 ± 0.01 units. (M ± σ), after incubation with Smecta - 0.57 ± 0.04 units; after GCM - 0.335 ± 0.005 units.

Example No. 6

The study of sorption from a solution of vitamin B 12 showed that 1 hour after incubation of 100 mg of sorbents with 1 ml of the drug, the concentration of vitamin in the solution with the Smecta sorbent was 184 ± 15 μg / ml, after HCM - 192 ± 8 μg / ml. The initial concentration of the drug in solution is 200 μg / ml.

The antitoxic properties of the drugs were evaluated in experiments on rats with radiation-thermal damage (RTP), causing a strong development of toxemia in the affected body. Experiments on the study of toxemia in RTP were carried out on Wistar rats weighing 200-220 g on a model of radiation-thermal damage, characterized by a pronounced aggravation of outcomes compared with the consequences of isolated radiation or burns. Animals were irradiated in a dose of 7.5 Gy, 2-5 pcs. in a plastic perforated container. Gamma irradiation of rats was carried out on Luch facilities with a source of ⁶⁰ Co. The dose rate was 1.0-1.5 Gy / min. A thermal burn of 15% of the body surface of the III-B degree was caused by the action of light radiation of 550 kJ / m ² quartz halogen incandescent lamps on a pre-trimmed back skin surface. The exposure time was 4.5 seconds.

The substance at a dose of 200 mg / kg in a volume of 0.5 ml of water was administered to rats 2 times during the first 2 days after exposure. To determine the binding ability of GKM and Smecta with respect to microbes located in the small and large intestines of rats, 1 cm of the cut intestine was placed in 5 ml of sterile saline solution, shaken for 30 seconds, transferred to a sterile tube and diluted by plating on culture medium in plates Petrie.

Example No. 7

The results of studies of changes in the bacterial content of E. Coli B / r in the small intestine of rats with RTP showed that 5 ± 0 · 10 ³ bacteria were in the biocontrol in 1 cm of the small intestine of rats, after RTP their content increased to 23 ± 8 · 10 ³ bacteria , the introduction of Smecta and GCM reduced the bacterial content to 11 ± 1 · 10 ³ and 5 ± 4 · 10 ^3, respectively.

Example No. 8

The introduction of enterosorbents also led to a decrease in the bacterial content of E. Coli B / r in the colon of rats after CRTP. 48 hours after CRT, the amount of E. Coli B / r increased on average from 50 · 10 ³ in 1 cm of intestine to 90 · 10 ³ , i.e. 1.8 times. The use of Smecta enterosorbent (France) reduced the content of E. Coli B / r in 1 cm of the intestine of affected rats to 27 · 10 ³ - 1.85 times less than normal and 3.3 times less than after CRT without treatment. The introduction of HCM caused an even greater decrease in the bacterial content of E. Coli B / r in 1 cm of the colon of rats: on average, up to 0.7 · 10 ³ or 71 times less than normal and 128 times less compared to CRT without the use of enterosorbents.

Example No. 9

A study of the development of histiogenic toxemia in affected animals revealed its increase 48 hours after RTP. The number of medium-weight molecules (MSM) increased from 1.35 ± 0.15 g / L (M ± σ) in rats of the control group to 1.78 ± 0.20 g / L after exposure (P <0.05).

The introduction of an aqueous suspension of the declared enterosorbent in the affected animals at a dose of 200 mg / kg led to a decrease in the concentration of MSM to 1.27 ± 0.07 g / l, i.e. almost to the level of intact rats.

Example No. 10

After RTP, the amount of toxic oligopeptides (TOP) in rat serum increased almost twofold: from 0.261 ± 0.049 g / l to 0.569 ± 0.073 g / l. The introduction of the declared enterosorbent led to the normalization of the level of oligopeptides - 0.331 ± 0.061 g / l - differences with the biocontrol group are unreliable (M ± σ).

Example No. 11. Reduction of alcohol intoxication

Outbred white mice weighing 20-26 g were injected intraperitoneally with 40 ° ethanol at the rate of 0.1 ml per 10 g of body weight; the mice then slept in the “on the side” position 60 ± 2 minutes (M ± σ). Administration of the inventive enterosorbent in a dose of 200 mg / kg by mouth 2-5 minutes after intraperitoneal administration of alcohol led to a decrease in sleep duration to 44 ± 5 minutes, i.e. GCM significantly reduced alcohol intoxication, determined by the decrease in the duration of sleep of mice in the "on the side" position.

Example No. 12. Reduction of the hypnotic effect of Nembutal

Outbred white mice weighing 21-26 g were injected intraperitoneally with a solution of nembutal (ethaminal sodium) at a dose of 30 mg / kg. Mice after injection of Nembutal slept in the "on the side" position 35 ± 3 minutes (M ± σ). Another group of mice 3-5 minutes after intraperitoneal administration of Nembutal was introduced through the mouth a solution of the inventive enterosorbent at a dose of 200 mg / kg The sleep duration of mice of this group was 27 ± 2 minutes (M ± σ), which indicates that GKM reduces the hypnotic effect of Nembutal, interrupting the metabolic processes of the drug in the body.

The above examples show that the problem is solved. The claimed enterosorbent effectively binds bacteria and reduces their content in the intestine, reduces the concentration of MSM and TOP in the blood serum, and reduces alcohol and drug intoxication.

Claims (2)

1. Enterosorbent based on silica-containing mineral raw materials, characterized in that it contains sedimentary opal-cristobalite rock of the Zikeevsky deposit of the Kaluga region, containing not more than 15 wt.% Clay minerals of the montmorillonite group and not more than 10 wt.% Minerals of the mica group . 2. Enterosorbent according to claim 1, characterized in that it has a degree of grinding of 0.1-1.0 mm