Magnetically controlled sorbent for endo- and exotoxin elimination in humans

FIELD: medicine.

SUBSTANCE: magnetite surface is modified with a compound forming a stable bond to a carrier particle by using surface-active groups giving the selectivity properties, and presented in the form of a coating of normal hydrocarbon chains C12H25 attached to a nucleus by a sulphide bond Fe-S; the above compound providing the bond of iron to the hydrocarbon chain is dodecyl mercaptan. Magnetite is prepared of ferric chloride (II) and ferric chloride (III) by chemical deposition with the concentration of 1 wt % in the solution and a mole ratio of iron (II) and (III) = 2.5, or of ferric sulphate FeSO4 (II) with adding a mixture of aqueous solution of potassium nitrate KNO3 and potassium hydroxide KOH at 3 moles of KOH and 0.1 mole of KNO3 per 1 mole of FeSO4.

EFFECT: invention provides the sorbent that is used to achieve the effective control of hazardous ingredients of blood plasma.

2 cl, 2 tbl

 

The invention relates to medicine and can be used in the treatment of endotoxemia, which accompany most urgent surgical diseases. According to most researchers, under the endotoxicosis (endogenous intoxication) refers to the manifestation of various pathological conditions, which is the basis of effects on the body of toxic products formed therein. The development of endotoxemia begins with receipt of toxins from the primary site of toxicity in the blood, lymph, interstitial space and spreading the blood around the body. If the primary source of endogenous intoxication cannot be removed surgically, and the protective system of the body is not able to neutralize the increasing flow of toxins, then there is a further progression of endotoxemia. In this case, only the additional, faster by removing toxins from the body able to resolve the critical situation. One such method is extracorporeal detoxification using filtration and sorption technologies. In this invention it is proposed to use magnetic sorbents in the form of suspensions of nano - and micro-size particles particles coated with a special molecular shells DL the biocompatibility and implementation of target sorption of toxins on the surface of the particles.

The known method of hemosorption on carbon sorbents for the treatment of diseases of the hepatobiliary system in which carbon hemosorbent has on its surface the volume of mesopores 0,17-0,35 cm3/g, micropore volume of 0.03 to 0.05 cm3/g and macropores 0.06 to 0.10 cm3/g when the total volume of all pores 0,26-0,50 cm3/g, and contains oxygen functional groups in the amount of 0.04-0.05 mEq./g [1].

The disadvantage of this technical solution is the use of sorbent, non-magnetic properties, which does not allow to achieve high rates of sorption efficiency in relation to remove toxic ingredients from the plasma of the human body.

Known, in addition, the magnetic sorbent prepared on the basis of micro - and nanoparticles of magnetite coated with polymerized silicic acid containing specific ligands [2].

However, this sorbent is intended solely for the adsorption of nucleic acids and can not provide efficient removal of endo - and exotoxins from the human body, such as total cholesterol, urea, creatinine and other toxins.

The applicant has set itself the practical task of developing the magnetic sorbent for removal of endo - and exotoxins from the human body, which is achieved effective control over the OC is ω harmful ingredients plasma, requiring mandatory output from the human body. A positive result was achieved due to a new essential features of the claimed magnetic sorbent for removal of endo - and exotoxins from the human body, is recorded in the following claims: "the magnetic sorbent for removal of endo - and exotoxins from the human body, made from nanoparticles of magnetite Fe3O4obtained on the basis of the chlorides of bivalent iron (II) and ferric chloride (III) or derived from ferrous sulfate FeSO4(II) adding the mixture of aqueous solutions of potassium nitrate KNO3and potassium hydroxide KOH, the surface of magnetite modified bifunctional compounds, forming a strong bond with the particle carrier through the surface-active groups, giving properties of selectivity and executed as a shell of normal hydrocarbon chains C12H25attached to the core by sulfide Fe-S, and the binding of iron with carbon chain carried out by a mercapto-group SH-R; the size of nanoparticles of magnetite Fe3O480÷150 nm; the magnetite nanoparticles of Fe3O4on the basis of ferric sulfate FeSO4(II) is added a mixture of aq is x solutions of potassium hydroxide KOH and aqueous solutions of potassium nitrate KNO 3taken at the rate of 3 mol of KOH and 0.1 mol KNO31 mol FeSO4(II); the bearer of the mercapto-group SH-R is dodecylmercaptan, providing the binding of iron with carbon chain; 1 g of a suspension of nanoparticles of magnetite in 50 ml of diphenyl ether is added 0.2 g of dodecylmercaptan".

Implementation of the invention is as follows.

Synthesis of magnetic carriers is carried out in two ways. Method No. 1 was based on the method of chemical deposition of salts of bivalent and trivalent iron. Why in the reactor is placed aqueous solutions of chlorides of iron (II) and (III) their concentration in solution of 1% of the mass. and the molar ratio of iron (II) and (III) = 2,5, which is freed from dissolved oxygen by passing nitrogen for 20 minutes.

Synthesis of magnetite is carried out by adding 8% of the mass. hydrate of ammonia dropwise to the reaction solution at room temperature and vigorous stirring. For complete formation of nanoparticles of magnetite Fe3O4the slurry is mixed for another 20÷40 minutes. The obtained magnetic dispersion is washed by centrifugation or sedimentation on the magnet with repeated washing with distilled water to lower the pH of the wash water is ~7÷8. The size of nanoparticles of magnetite Fe3O480÷150 nm.

Method 2 is based on Sul the ATA iron FeSO 4(II). When this aqueous solution of ferric sulfate FeSO4(II) is heated to 90°C in argon atmosphere. Then add drops with vigorous stirring a mixture of aqueous solutions of potassium nitrate KNO3and potassium hydroxide KOH at the rate of 3 mol of KOH and 0.1 mol KNO31 mol of ferrous sulfate FeSO4(II). Suspension with a black precipitate leave much to be mixed at the temperature of the reaction mixture to 90°C in argon atmosphere for two hours to complete crystallization of the particles of ferric sulfate FeSO4(II). The obtained magnetite was washed with water, then isopropyl alcohol and dried in vacuum at room temperature.

To make the surface of the particle extraction properties when using the magnetite nanoparticles obtained by methods 1 and 2, form the shell of a normal hydrocarbon chains With12H25attached to the core by sulfide Fe-S, and the binding of iron with carbon chain is realized by means of mercapto-group SH-R. then prepare a suspension of 1 g of the magnetite nanoparticles in 50 ml of diphenyl ether, treating the mixture with ultrasound for 30 minutes. In the resulting mass is added 0.2 g of dodecylmercaptan and boil at a temperature of 260°C for one hour with stirring. At the end of the process the product is separated by centrifugation and washed with ethanol and or what propanolol (number of experiments, respectively, D-1 and D-2).

The study of sorption properties of nanosorbent were performed on blood plasma and lymph for 12 people (donors). Bioliquids were given the 3rd Central military clinical hospital named A.A. Vishnevsky (3 CVKG them. A.A. Vishnevsky). As controlled criteria values of sorption capacity used indicators of content in blood plasma and lymph of their basic regular ingredients, the concentration of which you can judge the state of the cell, organ and overall homeostasis in various diseases.

To study the sorption was preparing a suspension of the modified magnetite in physiological solution with a concentration of 12.0±3.0 mg/ml, which were treated by ultrasound for 40 minutes in an ultrasonic bath Elmasonic-S-100 H at a frequency of 37 kHz for disaggregation of the particles and their distribution within the volume. Then mixed with 0.5 ml each of the resulting suspension with 3 ml of blood plasma. The resulting mixture was kept under constant shaking for 3 minutes and then separated particles by centrifugation at 2500÷3000 rpm for 15 minutes.

Controlled performance obtained with this technology the magnetic sorbent was following parameters.

ALT (alanine aminotransferase) and AST (aspartate aminotransferase) - specialized proteins (enzymes), which contains NR the three cells of the body and are involved in the metabolism of amino acids (substances, of which are proteins). Chol (total cholesterol) - source education in mammals bile acids, corticosteroids, sex hormones, vitamin D3. UREA (urea) - the major end product of nitrogen metabolism, quantitative determination of blood and urine is important for the diagnosis of pathological conditions of the liver and kidneys. Crea (creatinine) - the end product of protein metabolism, which is formed in the muscles and then is released into the blood. BiliT (total bilirubin) is the result of the breakdown of hemoglobin, myoglobin and cytochromes in the cells of the reticuloendothelial system, spleen and liver. GluC (glucose) - the main indicator of carbohydrate metabolism. TP (total protein) is the most important component of protein metabolism in the body, characterized by the total concentration of albumin and globulin in the blood serum. AlbG (albumin) is the most representative part of blood plasma proteins, plays a significant role in maintaining colloid osmotic pressure in the blood and is important for the body reserve of amino acids.

Analysis nasadochnykh bioliquids were conducted in the biochemical laboratory 3 CVKG them. A.A. Vishnevsky. Content controlled ingredients in the blood before and after adsorption was determined at the laboratory of biochemical analyzers Olympus (Germany) Advia 1200 (USA - Germany) and the system wedge is ical capillary electrophoresis Paragon (USA).

Table 1
The sorption efficiency of the modified magnetite in relation to the ingredients of the blood plasma (averaged relative to the initial concentration)
Ingredients plasmaExperience D-1Experience D-2
AST5612
ALT14□14
UREA5019
Crea4123
BiliT5033
GluC4923
TR5025
AlbG5320

Table 2
The sorption efficiency of the modified magnetite in relation to the ingredients of the blood plasma (averaged relative to the initial concentration)
Ingredients plasmaExperience D-1Experience D-2
AST3311
ALT□16□14
UREA1915
Crea234
BiliT3515
GluC235
TR318
AlbG207

Sources of information

1. Description of the invention to the patent of the Russian Federation "Method of hemosorption on carbon sorbents for the treatment of diseases of the hepatobiliary system No. 2343926, CL A61K 33/44, A61P 1/16, declared 10.09.2007, published 20.01.2009.

2. U.S. patent No. 8206990, class G01N 30/56, published 2012

p> 3. Description of the invention to the patent of the Russian Federation "Method of treatment of mesoporous carbon hemosorbent" No. 2362733, CL C01B 31/08, B01J 20/20, declared 19.05.2008, published 27.07.2009.

4. U.S. patent No. 2007/0105094 A1, class C72Q 1/70 (435/5), published 10.05.2007,

5. Description of the invention to the patent of the Russian Federation "Magnetic sorbent and method thereof" No. 2255800, CL A61M 1/36, declared 19.05.2008, published 27.07.2009.

6. Description of the invention to the patent of the Russian Federation "Magnetic sorbent, the manufacturing method and the method of its application" No. 2356620, CL B01J 20/06 declared 23.04.2008 published 27.05.2009.

7. Description of the invention to the patent of the Russian Federation "Porous magnetic sorbent No. 2226126, CL B01J 20/16 declared 30.12.2002, published 27.03.2004.

8. Description of the invention to the patent of the Russian Federation "Porous magnetic sorbent No. 2241537, CL B01J 20/26, declared 09.04.2003 published 10.12.2004.

1. The magnetic sorbent for removal of endo - and exotoxins from the human body, made from nanoparticles of magnetite Fe3O4received iron chloride (II) and iron chloride (III) by chemical vapor deposition with their concentration in solution of 1% of the mass. and the molar ratio of iron (II) and (III) = 2.5 or iron sulfate FeSO4(II) adding the mixture of aqueous solutions of potassium nitrate KNO3and potassium hydroxide KOH at the rate of 3 mol KOH and 0.1 mol KNO31 mol FeSO4while the surface of the magnetite modifica avana connection forming a strong bond with the particle carrier through the surface-active groups, giving properties of selectivity and executed as a shell of normal hydrocarbon chains C12H25attached to the core by sulfide Fe-S, and as mentioned compounds, providing the binding of iron with carbon chain selected dodecylmercaptan.

2. The sorbent according to claim 1, in which the size of the nanoparticles of magnetite Fe3O4is 80-150 nm.



 

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