RU2516961C1 - Magnetically controlled sorbent for endo- and exotoxin elimination in humans - Google Patents

Abstract

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

Description

The invention relates to medicine and can be used in the treatment of endotoxemia, which accompany most urgent surgical diseases. According to most researchers, endotoxemia (endogenous intoxication) refers to the manifestation of various pathological conditions, the basis of which is the effect on the body of toxic products formed in itself. The development of endotoxemia begins with the release of toxins from the primary focus of intoxication into the blood, lymph, interstitial space and their further spread by blood flow throughout the body. If the primary source of endogenous intoxication cannot be eliminated surgically, and the body's defense systems are not able to neutralize the increasing flow of toxins, then further progression of endotoxemia occurs. In this case, only an additional, faster-paced removal of toxins from the body is able to resolve the critical situation. One of these methods is extracorporeal detoxification using filtration and sorption technologies. The present invention proposes the use of magnetically controlled sorbents in the form of suspensions of nano- and micro-sized particles coated with special molecular shells for biocompatibility and the implementation of targeted sorption of toxins on the surface of the particles.

A known method of hemosorption on carbon sorbents for the treatment of pathologies of the hepatobiliary system, in which the carbon hemosorbent has on its surface a mesopore volume of 0.17-0.35 cm ³ / g, a micropore volume of 0.03-0.05 cm ³ / g and 0 , 06-0.10 cm ³ / g with a total volume of all pores of 0.26-0.50 cm ³ / g, and also contains oxygen functional groups in an amount of 0.04-0.05 meq / g [1].

The disadvantage of this technical solution is the use of a sorbent that does not have magnetic properties, which does not allow to achieve high indicators of its sorption efficiency in relation to removed toxic ingredients from the plasma of the human body.

Also known is a magnetically controlled sorbent prepared on the basis of micro- and nanoparticles of magnetite coated with polymerized silicic acid containing certain ligands [2].

However, this sorbent is intended solely for sorption of nucleic acids and cannot ensure the effective removal of endo- and exotoxins from the human body, for example, total cholesterol, urea, creatinine and other toxins.

The applicant set himself the practical task of developing a magnetically controlled sorbent for removing endo- and exotoxins from the human body, with the help of which effective control over a number of harmful blood plasma ingredients, requiring their mandatory removal from the human body, is achieved. The obtained positive result was achieved due to a new set of essential features of the inventive magnetically controlled sorbent for removing endo- and exotoxins from the human body, recorded in the following claims: “magnetically controlled sorbent for removing endo- and exotoxins from the human body, prepared from magnetite Fe ₃ O nanoparticles ₄ obtained based on the ferrous chloride (II) and ferric chloride (III) or obtained based on iron sulphate FeSO ₄ (II) with the addition of mixture of aqueous solutions of potassium nitrate KNO ₃ and potassium hydroxide KOH, and the surface of magnetite modified bifunctional compounds which form a strong bond with the carrier particles due to the surfactant groups conferring properties of selectivity and formed as a shell of the normal hydrocarbon chains of C ₁₂ H ₂₅ attached to the core via a sulfide bond Fe-S, and the binding of iron to the carbon chain is carried out through the mercapto group SH-R; the size of magnetite Fe ₃ O ₄ nanoparticles is 80 ÷ 150 nm; upon receipt of Fe ₃ O ₄ magnetite nanoparticles based on iron sulfate FeSO ₄ (II), the added mixture of aqueous solutions of potassium hydroxide KOH and aqueous solutions of potassium nitrate KNO _{3 was} taken from the calculation of 3 mol KOH and 0.1 mol KNO ₃ per 1 mol FeSO ₄ (II) ; the carrier of the mercapto group SH-R is dodecyl mercaptan, which provides binding of iron to the carbon chain; in 1 g suspension of magnetite nanoparticles in 50 ml of diphenyl ether add 0.2 g of dodecyl mercaptan. "

The implementation of the invention is as follows.

The synthesis of magnetic media is carried out in two ways. Method No. 1 was based on the method of chemical precipitation of salts of ferric and ferric. For this purpose, aqueous solutions of iron (II) and (III) chlorides are placed in the reactor with their concentration in the solution of 1% of the mass. and a molar ratio of iron (II) and (III) = 2.5, which are freed from dissolved oxygen by passing nitrogen for 20 minutes.

The synthesis of magnetite is carried out by adding 8% of the mass. ammonia hydrate dropwise to the reaction solution at room temperature with vigorous stirring. For the complete formation of magnetite Fe ₃ O ₄ nanoparticles, the suspension is left to mix for another 20–40 minutes. The obtained magnetic dispersion is washed by centrifugation or by deposition on a magnet with repeated washing with distilled water until the pH of the washing water decreases, equal to ~ 7 ÷ 8. The size of the magnetite nanoparticles Fe ₃ O ₄ is 80 ÷ 150 nm.

Method No. 2 is implemented on the basis of iron sulfate FeSO ₄ (II). In this case, an aqueous solution of iron sulfate FeSO ₄ (II) is heated to 90 ° C in an argon atmosphere. Then, a mixture of aqueous solutions of potassium nitrate KNO ₃ and potassium hydroxide KOH (3 mol KOH and 0.1 mol KNO ₃ per 1 mol of iron sulfate FeSO ₄ (II)) is added dropwise with vigorous stirring. The suspension with the black precipitate was allowed to mix at a temperature of the reaction mixture of 90 ° C in an argon atmosphere for two hours until the particles of iron sulfate FeSO ₄ (II) completely crystallized. The magnetite obtained is washed with water, then with isopropyl alcohol and dried in vacuo at room temperature.

To impart extraction properties to the surface of the particles using magnetite nanoparticles obtained by methods No. 1 and No. 2, a shell is formed of normal C ₁₂ H ₂₅ hydrocarbon chains attached to the core via a Fe-S sulfide bond, and iron is bonded to the carbon chain through mercapto -groups SH-R. Then a suspension is prepared in 1 g of magnetite nanoparticles in 50 ml of diphenyl ether, treating the mixture with ultrasound for 30 minutes. To the resulting mass add 0.2 g of dodecyl mercaptan and boil at a temperature of 260 ° C for one hour with stirring. At the final stage of the process, the product is separated by centrifugation and washed with ethanol or isopropanol (test numbers, respectively, D-1 and D-2).

A study of the sorption properties of nanosorbents was carried out on blood and lymph plasma for 12 people (donors). Biofluids were provided by the 3rd Central Military Clinical Hospital named after A.A. Vishnevsky (3 Central Exhibition Hall named after A.A. Vishnevsky). As controlled criteria, the values of sorption capacity used indicators of plasma and lymph levels of their main constant ingredients, the level of concentration of which can be used to judge the state of cellular, organ and general homeostasis in various diseases.

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

The following indicators were controlled indicators of the magnetically controlled sorbent obtained by this technology.

ALT (alanine aminotransferase) and AST (aspartate aminotransferase) are special proteins (enzymes) that are contained within the cells of the body and are involved in the metabolism of amino acids (the substances that make up proteins). Chol (total cholesterol) is the source of the formation in the body of mammals of bile acids, corticosteroids, sex hormones, vitamin D ₃ . UREA (urea) is the most important end product of nitrogen metabolism, the quantitative determination of which in the blood and urine is important for the diagnosis of the pathological condition of the liver and kidneys. Crea (creatinine) is the final product of protein metabolism, which is formed in the muscles and then secreted into the blood. BiliT (common bilirubin) - is formed as a result of the breakdown of hemoglobin, myoglobin and cytochromes in the cells of the reticuloendothelial system, spleen and liver. GluC (glucose) is 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 globulins in the blood serum. AlbG (albumin) - the most representative part of blood plasma proteins, plays a significant role in maintaining colloid osmotic pressure in the blood and serves as an important reserve of amino acids for the body.

Analysis of supernatant bioliquids was carried out in the biochemical laboratory of 3 TsVKG im. A.A. Vishnevsky. The content of controlled ingredients in the blood before and after sorption was determined on laboratory biochemical analyzers Olympus (Germany) Advia 1200 (USA - Germany) and Clinical Capillary Electrophoresis System Paragon (USA).

Table 1 Sorption effectiveness of modified magnetite with respect to blood plasma ingredients (in averaged percent relative to the initial concentration) Plasma ingredients Experience D-1 Experience D-2 AST 56 12 ALT fourteen □ 14 UREA fifty 19 Crea 41 23 Bilit fifty 33 Gluc 49 23 TR fifty 25 Albg 53 twenty

table 2 Sorption effectiveness of modified magnetite with respect to blood plasma ingredients (in averaged percent relative to the initial concentration) Plasma ingredients Experience D-1 Experience D-2 AST 33 eleven ALT □ 16 □ 14 UREA 19 fifteen Crea 23 four Bilit 35 fifteen Gluc 23 5 TR 31 8 Albg twenty 7

Information sources

1. Description of the invention to the patent of the Russian Federation “Method of hemosorption on carbon sorbents for the treatment of pathologies of the hepatobiliary system” No. 2343926, cl. A61K 33/44, A61P 1/16, claimed September 10, 2007, published January 20, 2009.

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

3. Description of the invention to the patent of the Russian Federation “Method for processing carbon mesoporous hemosorbent” No. 2362733, class. C01B 31/08, B01J 20/20, claimed 05/19/2008, published 07/27/2009.

4. US patent No. 2007/0105094 A1, class C72Q 1/70 (435/5), published 05/10/2007.

5. Description of the invention to the patent of the Russian Federation “Magnetically controlled sorbent and method for its preparation” No. 2255800, class. A61M 1/36, claimed 05.19.2008, published 07.27.2009.

6. Description of the invention to the patent of the Russian Federation “Magnetically controlled sorbent, method of its manufacture and method of its application” No. 2356620, class. B01J 20/06, claimed 23.04.2008, published 05.27.2009.

7. Description of the invention to the patent of the Russian Federation "Porous magnetic sorbent" No. 2226126, class. B01J 20/16, claimed December 30, 2002, published March 27, 2004.

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

Claims (2)

1. Magnetically controlled sorbent for removal of endo- and exotoxins from the human body, prepared from nanoparticles of magnetite Fe ₃ O ₄ , obtained iron (II) chloride and iron (III) chloride by chemical precipitation with a concentration of 1% in solution. and a molar ratio of iron (II) and (III) = 2.5 or iron sulfate FeSO ₄ (II) with the addition of a mixture of aqueous solutions of potassium nitrate KNO ₃ and potassium hydroxide KOH at the rate of 3 mol KOH and 0.1 mol KNO ₃ per 1 mol of FeSO ₄ , while the surface of magnetite is modified by a compound that forms a strong bond with the carrier particle due to surface-active groups imparting selectivity properties and made in the form of a shell of normal C ₁₂ H ₂₅ hydrocarbon chains attached to the core via a Fe– sulfide bond S, moreover, as said compound, providing which enhances the binding of iron to the carbon chain, dodecyl mercaptan is selected. 2. The sorbent according to claim 1, in which the size of the magnetite Fe ₃ O ₄ nanoparticles is 80-150 nm.