RU2233661C1 - Selenium-terpene-protein organic substance eliciting biologically active effect - Google Patents

Abstract

FIELD: clinical medicine, biochemical pharmacology.

SUBSTANCE: invention relates to a substance that can be used as an anti-inflammatory, wound-healing, antihemorrhagic, anticancer, anthelmintic and cardiotonic agent. Selenium-terpene-protein is a substance that elicits the broad spectrum of biological effect and comprises selenoprotein, selenium and sesquiterpene lactone. The substance is prepared by effect of electromagnetic high-frequency field with power 5 x 10 Wt/cm and frequency 20-30 MHz. Selenium-terpene-protein comprises additionally ketone thujone, i. e. selenothujonoterpenoprotein or bromide anion, i. e. selenium-terpene-protein bromide, or terpine hydrate - selenium-terpine-terpene-protein or alpha-pinene - selenium-pinene-terpene-protein, or azulene - selenium-azulene-terpene-protein. Proposed substances provide their using for both stimulation of immune system and for regulation of different metabolic processes.

EFFECT: valuable medicinal properties of substance.

6 cl, 13 dwg, 6 ex

Description

The invention relates to biochemical pharmacology and pharmacodynamics, biochemical medicine, cellular and molecular biology, immunopharmacology and applied medicine and can be used in clinical medicine as anti-inflammatory, wound healing, hemostatic, anti-cancer, anthelmintic and cardiotonic agent. This substance can be used both to stimulate the immune system and to regulate a variety of metabolic processes.

Currently, in clinical medicine, organic substances of natural origin are used to correct immune and metabolic processes, as well as to regulate cardiovascular mechanisms. These are, as a rule, preparations from extracts of goiter, lymph nodes, connective tissue and spleen. Such drugs include substances well known in clinical practice - thymogen, T-activin, myelopid, thymoptin, etc. (Mashkovsky MD Drugs that correct the processes of immunity. In the book. Medicines, Kharkov, “Torsing”, v.2, 1997, 199–207).

Recently, the attention of numerous researchers has attracted selenium and its compounds, the so-called selenium-containing proteins, because it is the biochemical functions of selenium that are determined not by the trace element itself, but by selenoprotein organic substances. The lack of this trace element can lead to a violation of cellular integrity, a change in the metabolism of thyroid hormones, the activity of biotransforming enzymes, an increase in the toxic effect of heavy metals, and an increase in the concentration of glutathione in plasma. A characteristic feature of selenium-containing mammalian proteins is that they are associated with redox processes that occur inside and outside the cell. To date, twelve selenium-containing proteins have been characterized, including enzymes such as glutathione peroxidase, iodothyronine deiodinase, thioredoxin reductase, selenophosphate synthetase, and others. The data of isotope analysis and the results of theoretical studies suggest that in the mammalian body there can be from 20 to 100 selenium-containing proteins (V.A. Tutelian et al. In the book Selenium in the human body, M., 2002, p. 18- 34).

To date, the biological functions of selenium have been fairly well studied, including the mechanism of action of glutathione peroxidase, the process of peroxynitrite recovery under the influence of selenium-containing proteins, the mechanism of interaction of selenium with antioxidants, especially vitamins C, E and A. The role of selenium in the occurrence and development of cardiac diseases has also been studied. , the effect of selenium on prostaglandin biosynthesis, the effect of selenium on reproduction, and the role of selenium in the metabolism of xenobiotics (V.A. Tutelian et al. in the book Selenium in the body Man. M., 2002, p. 88-117).

The presented data show that selenium and selenium-containing proteins are necessary biological components and regulators in the implementation of complex metabolic processes, including redox, vascular, neurohumoral and others. In this regard, the issue of obtaining such biologically active compounds of selenium, which could be directly involved in the normalization and restoration of a number of homeostatic processes, is particularly relevant. However, the selenium drugs obtained so far are either the microelement itself, or its compounds with vitamins, including amino acids, for example, cysteine or methionine. In this regard, the new class of medicinal substances being created must, to some extent in their structure and mechanism of action, correspond to selenium-containing proteins that directly carry out various physiological processes - metabolic, immunobiological, reproductive, gene-protective, and others.

To solve this problem, medicinal substances with the action of selenium-containing proteins are proposed. They include a low molecular weight component - a protein obtained by extraction from protein-containing plant materials with organic solvents, selenium, sesquiterpene lactone, thujone, bromine anions, terpinghydrate, pinene and verbenol, azulene.

The resulting funds are called selenoterpenoprotein organic substances. The invention is illustrated in figures 1-13.

Obtaining selenoterpenoprotein organic matter - STP

As the starting products use the green mass of protein-containing plants, such as cereals, legumes, corn leaves. By extraction from the plant mass, protein is recovered. The extraction is carried out with ether, ethyl alcohol, dimexide. The extract obtained is evaporated and lyophilized. The powder is dissolved in a 60-90% ethanol solution. Selenium obtained by thermolysis of selenium urea is introduced into the solution (from the calculation of 6-12 μg per 1 ml of solution). Then, a certain amount of sesquiterpene lactone is added to the mixture and placed in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz).

1. Substance selenoterpenoprotein - STP

The substance includes a molecule of selenoprotein (Fig. 1), to which sesquiterpene lactone - STL is attached (Fig. 2), to the carboxyl groups of the side chains - the residues of glutamic and / or aspartic acid and / or to the C-terminal carboxyl groups through complex ether linkage (FIG. 3). The selenoterpenoprotein molecule is two polypeptoid alpha and beta chains connected by a disulfide bridge at cysteine residues (the α chain consists of 20 amino acid residues). Selenium obtained by thermolysis of selenourea, enters the lateral alkyl radicals of amino acids, connecting some units of the alpha and / or beta chains of one or two neighboring protein molecules, thereby additionally “linking” them.

At all physiological pH values, the molecule of selenoterpenoprotein is ionized. The C- and N-terminal groups and the side chains bonded to α-carbon atoms containing a carboxyl or amino group (-COO ^- , -NH are charged $\genfrac{}{}{0ex}{}{\text{+}}{\text{3}}$ )

2. The substance selenotujonoterpenoprotein (Fig.6)

The substance includes an ionized molecule of selenoterpenoprotein (Fig. 3), to which a thujone-T ketone (Fig. 4) is attached via amino groups of the side chains of arginine residues and / or amino terminal groups (Fig. 5).

3. The substance bromide-selenoterpenoprotein (Fig.7)

The substance includes an ionized molecule of selenoterpenoprotein, to which bromine (Br ^- ) anions are attached at the amino groups of the side chains of arginine and / or histidine residues and / or amino end groups.

4. The substance selenoterpinoterpenoprotein (Fig.9)

The substance includes an ionized molecule of selenoterpenoprotein, to which terpin hydrate - TH (Fig. 8) is attached to the carboxyl groups of the side chains of glutamic and / or aspartic acid residues and / or to the C-terminal carboxyl groups via an ester linkage.

5. The substance selenopinenoterpenoprotein (11)

The substance includes an ionized molecule of selenoterpenoprotein (Fig. 3), to which alpha-pinene (P), pre-oxidized to verbenol (Fig. 10), is attached to the carboxyl groups of the side chains of glutamic and / or aspartic acid residues and / or to the C-terminal carboxyl groups through an ester linkage.

6. The substance selenoazulenoterpenoprotein (Fig.13)

The substance includes an ionized molecule of selenoterpenoprotein (Fig. 3), to which azulene (Az) is attached, substantially polarized (Fig. 12) at the terminal carboxyl (-) and amino - (+) groups and / or along the side chains of arginine (+ ), and / or histidine (+), and / or glutamic acid (-), and / or aspartic acid (-) (Fig. 13).

Examples of obtaining selenoterpenoprotein organic substances

Example 1

The method of obtaining selenoterpenoprotein

The required amount of sesquiterpene lactone is added to the ionized molecule of selenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of STL: selenoprotein 4: 1.

Example 2

The method of obtaining selenotujonoterpenoprotein

The required amount of thujone ketone is added to the ionized molecule of selenoterpenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of T: selenoterpenoprotein 4: 1.

Example 3

The method of obtaining bromide-selenoterpenoprotein

The required amount of bromine anions obtained by electrolysis of NaBr is added to the ionized molecule of selenoterpenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of Br: selenoterpenoprotein is 4: 1.

Example 4

The method of obtaining selenoterpinoterpenoprotein

The required amount of terpinghydrate is added to the ionized molecule of selenoterpenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of TH: selenoterpenoprotein 4: 1.

Example 5

The method of obtaining selenopinenoterpenoprotein

The required amount of pinene, previously oxidized to verbenol, is added to the ionized molecule of selenoterpenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of P: selenoterpenoprotein 4: 1.

Example 6

The method of obtaining selenoazulenoterpenoprotein

The required amount of azulene is added to the ionized molecule of selenoterpenoprotein. The mixture is kept in an electromagnetic high-frequency field (power 5 · 10 ^-3 W / cm, frequency 20-30 MHz). The molar ratio of Az: selenoterpenoprotein is 4: 1.

The acute toxicity of the drug was studied with intraperitoneal administration of Se-TP in a dose of 250 mg / kg to mice (age 6 weeks, weight 28-30 g). The LD value was 700 mg / kg.

In a study of total toxicity, Se-TP was administered to mice at a dose of 100 mg / kg per day intraperitoneally continuously for 10 days. No weight loss or any other abnormalities were noted, and subsequently, when these mice were observed for three months, no abnormalities were found.

As a result of the studies, it was found that Se-TP is an organic compound, has a pronounced biological effect, is non-toxic, completely soluble in water and biological fluids - blood, lymph and cerebrospinal fluid. 1 ml of Se-TP contains, depending on the amount of amino acids in the ligand, from 20 to 30 mg of the substance and 8-10 μg of selenium.

The dose of the drug administered depends on the clinical condition of the patient, his age, weight, as well as the route of administration. An effective therapeutic daily dose for a patient is from 6 to 150 mg of active substance, which is administered simultaneously or fractionally.

The method of activation and stimulation of metabolic and immune processes is as follows.

The obtained preparation Se-TP is a solution of the active substance in 60-90% ethanol, 1 ml of which contains 20-30 mg of the drug substance. Se-TP is administered orally, intramuscularly, intravenously, intraarterially, and also externally as a suspension in an oil solution or as an ointment with an oil-alcohol filler.

When administered orally, the required dose is calculated as follows:

20 drops of the solution contain 20-30 mg of the drug substance;

10 drops (1/2 ml) contain 10-15 mg of the drug;

5 drops (1/4 ml) contain 5 -7.5 mg of the drug;

3 drops (1/7 ml) contain 3.3 mg of the drug.

With intramuscular, intravenous or intraarterial administration, the drug is dissolved in physiological saline for better dissociation in a ratio of 1: 10-1: 20 using 60% alcohol or 1:15 with 90% alcohol.

For example, for moderate clinical pathology, the drug is administered once as an injection from 0.1 to 0.5 ml per day, for severe pathology, the specified dose is administered twice a day. During intensive therapy, the drug is administered intramuscularly or intravenously (intraarterially) in 1.0-2.0 ml, respectively, with the addition of 10 or 20 ml of physiological saline. In the case of oral administration, depending on age, weight and degree of damage, from 0.5 to 3.0 ml, moreover, the drug is dissolved in 1/3 cup boiled slightly warm water (36-38 ° C). The duration of treatment can vary from 1 to 3 months, in the case of a severe course of the disease, the treatment is repeated with an interval of 1-2 months until a pronounced clinical effect. During the treatment period, the effect of therapy is monitored by the study of biochemical parameters and immunological status.

Claims (6)

1. Selenoterpenoprotein organic substance with a wide spectrum of biological action, characterized in that it contains selenoprotein, which is two polypeptide α and β protein chains, into the side alkyl radicals of which selenium is introduced, connecting some units of the α or β chains of one or two adjacent protein molecules, the polypeptide chains are also connected by a disulfide bridge at the cysteine residues, while the selenoprotein is connected to the sesquiterpene lactone via the carboxyl groups of the side chains and residues of glutamic and / or aspartic acid and / or C-terminal carboxyl groups and obtained under the influence of an electromagnetic field with a power of 5 · 10 ^-3 W / cm at a frequency of 20-30 MHz. 2. The substance according to claim 1 further comprises a thujone ketone and is selenotujonoterpenoprotein (Fig.6), while the thujone with selenoterpenoprotein is connected through the amino groups of the side chains of arginine residues and / or the amino end groups. 3. The substance according to claim 1 additionally contains bromine anions and is bromide-selenoterpenoprotein (Fig. 7), while bromine with selenoterpenoprotein is connected via amino groups of the side chains of arginine and / or histidine residues and / or terminal amino groups. 4. The substance according to claim 1, additionally contains terpin hydrate and is selenoterpinoterpenoprotein (Fig. 9), while terpinghydrate with selenoterpenoprotein is connected via carboxyl groups of the side chains of glutamic and / or aspartic acid residues and / or C-terminal carboxyl groups. 5. The substance according to claim 1 additionally contains alpha-pinene, pre-oxidized to verbenol, and is selenopinenoterpenoprotein (11), while pinene with selenoterpenoprotein is connected via carboxyl groups of the side chains of glutamic and / or aspartic acid and / or C residues -terminal carboxyl groups. 6. The substance according to claim 1 additionally contains azulene and is selenoazulenoterpenoprotein (Fig.13), while azulene is connected to selenoterpenoprotein at the terminal carboxyl (-) and amino (+) groups and / or side chains of arginine (+) and / or histidine (+), and / or glutamic acid (-) and / or aspartic acid (-).

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