The method of obtaining nanomolecular selenoprotein with immunotropic and metabolic action
The invention relates to biopharmacology and comes to obtain the aimed at restoring metabolic processes and correction functions of the immune system. The method of obtaining Nanomolecular selenoprotein includes the extraction of protein from belkovosoderzhaschie plant materials with an organic solvent followed by the addition of selenium, processing the mixture in a sinusoidal electromagnetic high-frequency field, then the trypsin by adding to the formed nanoparticles molecules selenoprotein selenium ions in the ratio 1:8 and re-conditioning in high-frequency electromagnetic field 20-30 GHz. The means obtained by the proposed method has a strong immunotropic and metabolic action, non-toxic, soluble in water and biological fluids - blood, lymph and cerebrospinal fluid. 1 N. and 3 C.p. f-crystals, 3 ill.
The invention relates to biopharmacology, immunopharmacology, biotechnology, clinical medicine and can be used to obtain a variety selenoproteins drugs aimed at restoring the immune system and correction of metabolic the La obtain a variety of selenium-containing proteins and selenium amino acids.
Currently, the problem with the exchange of selenium and its role in the formation of immune and metabolic processes, has received priority (C. A. tutelian and other Selenium in the human body”. M.: Izd. Russian Academy of medical Sciences, 2002, S. 11-45). In particular, it was found that the specific pathology caused by selenium deficiency develops in humans at the intake of selenium less than 19 mcg per day for men and 14 MGK women. Physiological levels of consumption and assimilation of selenium controlled by the amount of the proceeds of this trace element in the body and the activity of the enzyme glutathione peroxidase, which cause the formation of so-called selenium homeostasis (C. A. tutelian and other Selenium in the human body”. M.: Izd. Russian Academy of medical Sciences, 2002, S. 146-156). In this regard, in clinical medicine originated the whole area, based on the use of selenium as a trace element and amino acids connected with selenium. Specific and nonspecific mechanism of incorporation of selenium in macromolecules is regulated by a specific gene system as in bacteria and mammals. The incorporation of selenocysteine in special molecules splenopathy directly carried genes and special enzymes (Bing Lei Do What Is Selenium”, Boston, London, 2000; August By Side. “Selenium metabolism in bacteria”, Munich University, Germany, in the book “Selenium”. Boston, London, 2000). For example, the incorporation of selenocysteine in transfer RNA occurs only under the supervision of gene structures involving special enzymes.
The study of the complex mechanism of inclusion selenium amino acids in selenesetestcase proteins suggests that the methods of genetic engineering in this case may not be widely used for the production of a large number of selenium-containing protein drugs, because they require simultaneous participation in the process of matrix synthesis and transport of RNA. At the same time the need to develop medicines containing selenium, coupled with protein is extremely high. Recently created a number of technologies to get selenoproteins organic matter from amino acid chains connected by a selenium ions (RF Patent for the invention №2177323 - Selenoproteins organic substance having immunokorrigiruyuschy effect). This patent describes a technology for obtaining in vitro molecule selenoprotein.
However, this invention does not complete the technology created class of drugs selenoproteins substances. This technology does not provide for the possibility of additional saturation of protein selenium ions, which does not allow selenoprotein as the ligand to be of high energy activity. In addition, not solved the problem of intramolecular selenaria protein matrix. At the same time to get active ligand molecules selenoprotein - you must use Nanomolecular technology with simultaneous high-frequency effects in order to selenaria the inner surface of the molecule.
The creation of Nanomolecular selenoproteins substance in some degree must be identical to the synthesis of selenium-containing proteins in micro - and macroorganism. Given the role of selenium in the regulation of diverse processes immunotropic, and metabolic anticancer, it is necessary in the process to provide active saturation selenium ions most selenoproteins matrix.
To solve this problem is proposed a technology of obtaining Nanomolecular selenoproteins substance that has a physiological effect of selenium-containing proteins. Nanomolecular selenoproteins matrix includes discomolo, and selenium ions produced by the electrolysis of Selenomethionine. Along with this, for the implementation of the intramolecular selenization necessary protein enzyme (Nozdrachev A. D., editor of “the Beginning of physiology.”, Ed. SPb 2001, S. 892-893) splitting the molecules on the nanoparticles with the subsequent addition of selenium ions and the use of ultra-high gigahertz frequencies.
The method of obtaining Nanomolecular selenoprotein is as follows (Fig.1-3).
The first stage of obtaining Nanomolecular selenoproteins substances (Fig.1).
Synthesis selenoproteins matrix
As the source material used green mass of protein-enriched plants, such as cereals, legumes, leaves of corn and other pre-dried and crushed. By extraction with ether, ethyl alcohol, or Dimexidum from the received raw vegetable protein extract protein. The protein extract is evaporated and produce lyophilic drying; custom powder dissolved in 60-90% ethyl alcohol, then enter the selenium obtained by thermolysis of Selenomethionine, at the rate of 10-12 μg per 1 ml of solution and put the mixture in the electromagnetic (power flux-density 5·10-3W/cm2) sinusoid is peptideand-chain connected by a disulfide bridge at cysteine residues;the chain consists of 20 amino acids residues,- from 29 amino acids. Selenium is obtained by thermolysis of Selenomethionine, included in the side alkyl radicals of amino acids, connecting some linksand/or-chains of one or two neighboring protein molecules, thereby further stitching them (Fig.1). The molar ratio of protein and selenium 1:4.
The second stage of obtaining Nanomolecular selenoproteins substances (Fig.2).
Obtaining nanoparticles selenoproteins matrix
The obtained selenoprotein affect the enzyme trypsin in a dilution of 1:1000 (1/20 ml per 100 ml of the substance), which hydrolyzes links the carboxyl group of basic amino acids - in this case, arginine, included in theand-chain protein. This process is accompanied by the separation of molecules on selenoprotein nanostructures between the two amino acids arginine-chain and splitThe synthesis of nanoparticles selenoproteins matrix into a single structure with additional enrichment of the protein ions selenium
To the obtained nanoparticles molecules selenoprotein add an additional 10-12 mcg of selenium obtained by thermolysis of Selenomethionine, and this mixture is placed in a high-frequency electromagnetic field - 20-30 GHz duration of 5 minutes when the power flux-5·10-3W/cm2. In nanomolecule contains 20-24 µg of selenium, and optionally two diselenide communication between the two amino acids at the top and amino acids isoleucine and glutamine at the bottom. The molar ratio of protein and selenium in Nanomolecular selenoproteins matrix 1:8.
The obtained nano-molecules of selenoprotein consists of two polypeptideand-chain protein, connected by a disulfide bridge at cysteine residues, selenuim bonds between the amino acids valine and leucine, selenium bond between glutamic acid and two diselenide relationship betweenand-chain. When all physiological pH values nano-molecules of selenoprotein ionized. Charged operasie carboxyl or amino group (Fig.3).
According to C. A. Tutelyan and others (“Selenium in the human body”. M.: Izd. FRAMES, 2002, S. 11-45), all selenesetestcase proteins, including received Nanomolecular selenoprotein have active immunotropic and metabolic action. Research conducted recently a number of scientists (Century A. Kuprin, A. C. Rock and others, in the book “Immunopharmacology trace elements”, Ed. KMK; M, 2000, S. 14-61) show that selenium and selenoproteins compounds in humans and animals regulate the action biotransforming enzymes activate the metabolism of pituitary and thyroid hormones, as well as reduce the possibility of occurrence of various immunodeficiency processes in tumors, a variety of autoimmune and allergic diseases, block transcription and replication of the AIDS virus.
1. The method of obtaining Nanomolecular selenoprotein with corrective action functions of the immune system and metabolic processes, characterized in that the protein is obtained by extraction of protein-enriched plants organic solvents, freeze-dried and then dissolved in 60-90% ethyl alcohol and dobavlyaeti power 5·10-3W/cm2) sinusoidal high-frequency field 20-30 MHz for 5 min, the resulting substance is treated with trypsin, the resulting nanoparticles molecules selenoprotein add selenium ions, is kept in a high-frequency electromagnetic field 20-30 GHz for 5 minutes when power flow 5·10-3W/cm2when this molar ratio of protein and selenium is 1:8, and the obtained nano-molecules of selenoprotein is aand-chain protein, connected by a disulfide bridge at cysteine residues, selenuim bonds between the amino acids valine and leucine, as well as between glutamic acid and two diselenide relationship betweenand-circuit, while nano-molecules of selenoprotein ionized when all physiological pH values, are charged With - and N-terminal group and associated withcarbon atoms of the side chain containing a carboxyl or amino group.
2. The method according to p. 1, where as belkovosoderzhaschie vegetable raw materials use of cereals and leguminous plants, the leaves of the corn.
3. The method according to p. 1, the p. 1, where the substance of selenoprotein treated with trypsin based 1/20 ml of enzyme per 100 ml of the substance.
Currently, in the treatment of infectious-inflammatory, autoimmune and oncological diseases in clinical medicine is widely used medicines, which have immunomodulatory properties (I. B. Mikhailov
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