Pharmaceutical composition for therapy of acute toxic conditions

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, and concerns a pharmaceutical composition for the therapy of acute toxic conditions. The composition for the therapy of acute toxic conditions contains a protein - human lactoferrin, and additionally contains non-replicating nanoparticles with an inserted gene of human lactoferrin, and an expressing buffer in the following proportions per a dose: human lactoferrin 50 to 100 mg; non-replicating nanoparticles - 7×1011 of physical particles; the expressing buffer - the rest, ml. Human lactoferrin is either donor breast milk lactoferrin, or any human lactoferrin.

EFFECT: invention provides fast onset and prolonged antitoxic action.

4 cl, 8 ex, 2 dwg

 

The invention relates to medicine, in particular nanotechnology and toxicology, and can be used for the prevention and treatment of toxic conditions of various etiologies, including acute.

There is a method of treatment of postoperative complications using preparations containing protein antioxidants (RF Patent 2199337). In this patent the treatment of postoperative complications with symptoms of multiple organ failure is carried out by co-introducing into a patient a medicinal product containing human lactoferrin, and medicinal product containing ceruloplasmin and preparations containing human lactoferrin and ceruloplasmin person injected systemically daily intravenous drip in isotonic glucose solution or sodium chloride. In addition, the drug containing ceruloplasmin human, is administered systemically daily intravenous drip, and solution preparation containing human lactoferrin daily washed with festering wounds, cavities, and/or irrigate the respiratory tract.

Lactoferrin is a single-stranded metallovedeniye protein. It is well known that this natural protein has a number of medicinal properties, including bactericidal and bacteriostatic activity, participates in the regulation of cellular and humoral immunologic the fir reactions anti-inflammatory and other processes.

The disadvantage is the fact that the claimed preparation though, and operates immediately after the introduction, but only the first day, and then excreted from the body, which is a significant disadvantage in therapy because of the need for frequent introduction of drug therapies

Known antibacterial, antioxidant, detoxifying, immunomodulating and antineoplastic drug patent RF №2165769 containing lactoferrin person as the main active ingredient and pharmaceutically acceptable additives, the product contains mass. percent: lactoferrin man - of 10.0 to 90.0; pharmaceutically acceptable additives - the rest. The claimed medicinal product may be made in the form of a solution for internal administration, in the form of a solution for intracavitary or intravesical injection, solution for oral administration in the form of solution for treatment of wound surfaces, in the form of eye drops, solution for intranasal use, in the form of ointments, in the form of a bolus for oral administration in the form of suppositories for rectal or intravaginal use, in the form of tablets.

This patent selected by the authors for the prototype.

The disadvantage is that the claimed drug though and dei is there immediately after the introduction, but only the first day, and then excreted from the body, which is a significant disadvantage in therapy because of the need for frequent administration of the drug for the implementation of therapy.

The General disadvantages of the compositions and preparations and methods of treatment are:

1) difficulty in achieving sustained therapeutic effect due to the presence of active components isolated and purified protein lactoferrin, which in all ways to rapidly excreted from the body of the patient;

2) to maintain therapeutically effective concentration in the body the necessity of repeated administration of pharmaceutical compositions containing lactoferrin;

3) the cost of large quantities of the drug, medical instruments and time of the medical staff to achieve the desired result of the treatment;

4) for the compositions and preparations, which include lactoferrin derived from milk, uniqueness and scarcity of this type of raw materials severely limits the scale of production and, consequently, the possibility of its application in the required quantities by medical indications.

Also known recombinant human lactoferrin produced in various systems, including viral vectors carrying the gene for lactoferrin man, the which by their physical, biochemical and biological properties similar to native lactoferrin (Conzalez-Chavez, S. A. et al., Lactoferrin: structure, function and applications, Int. J. of Antimicrobial Agents, 2009, No. 33, s-308. - Lactoferrin : structure, function and application).

Known effective treatment induced mammary tumors in mice by administering to the tumor tissue recombinant adenovirus carrying the gene for human lactoferrin. This composition had a prolonged effect and were carried out once in two weeks (Wang J. et al., Inhibition of tumor growth by recombinant adenovirus containing human lactoferrin through inducing tumor cell apoptosis in mice bearing emt6 breast cancer, Arch.Pharm.Res., 2011, №34 (6), 987-995 - Inhibition of tumor growth with recombinant adenovirus carrying the gene of human lactoferrin by apoptosis of tumor cells in mice suffering from breast cancer EMT).

The disadvantage of this composition is the following. Expression of target protein lactoferrin using adenovectors structures in the body does not begin with the moment of introduction, and only after a few hours, which is a significant disadvantage in relation to the treatment of acute toxic effects, although time lactoferrin continues after a single injection of a long time, which gives advantages in the treatment of chronic toxicosis, but cannot be used for the treatment of acute toxic effects.

The technical is the task of the invention is aimed at creating a pharmaceutical composition with rapid onset and prolonged antitoxic action-based nanostructures, producing directly in the body of a human lactoferrin, and is suitable for the treatment of acute toxic conditions of various origins.

This problem is solved due to the fact that the pharmaceutical composition for the treatment of acute toxic state, containing the protein human lactoferrin further comprises rereplacenocase nanoparticles with the insertion of a gene of human lactoferrin, and defines the buffer. If this dose is 3 ml Dose of the inventive pharmaceutical composition contains:

lactoferrin man from 50 to 100 mg;

rereplacenocase nanoparticles - 7×1011physical particle;

formulating buffer - rest, Jr.

At the same time as lactoferrin human use lactoferrin donor breast milk or any human lactoferrin

Disclosure of inventions

The technical solution is implemented in the application due to the fact that we used the combination of the properties of drugs of native human lactoferrin derived from donor human milk, and human lactoferrin expressed nerealizirane nanoparticles on the basis of the genome of adenovirus 5-th serotype, with the insertion of exogenous DNA, including the gene encoding the protein human lactoferrin in the same pharmaceutical composition, which allows for therapy of acute oxicash conditions of various etiologies in a single composition. The composition also contains formulating the buffer as a pharmaceutically acceptable additive. This ensures efficient operation of lactoferrin almost since the introduction and prolong its therapeutic concentration of 28-30 days without the implementation of additional injection.

Technical, medical and economic results in the implementation of the claimed invention are achieved due to the fact that just as in the well-known drug on the basis of human lactoferrin isolated from breast milk, the main therapeutic agent is lactoferrin person, the level of which in the body immediately after administration of the pharmaceutical compositions according to the invention is provided native lactoferrin, and then produced nerealizirane nanoparticles recombinant lactoferrin.

The pharmaceutical composition according to the invention designed as a dosage form in the form

lactoferrin man from 50 to 100 mg;

rereplacenocase nanoparticles - 7×1011physical particle;

formulating buffer - rest, Jr.

and is used as a solution for intravenous injection.

The pharmaceutical composition is the starting product for the preparation of various dosage forms, the use of which is determined depending on the etiopathogenesis of toxicosis. Declare farmacevticheskaja composition based on native human lactoferrin and rereplacenocase nanoparticles with the insertion of a gene encoding human lactoferrin, passed preclinical and clinical trials to study specific (therapeutic) efficiency and General toxic effect, which demonstrated the safety of this composition and therapeutic activity as detoxifying substances at various toxic conditions, especially acute toxicosis, which is illustrated by the following examples.

The examples below show:

- designing rereplacenocase nanoparticles and their enhanced in the required amount;

- creation of the pharmaceutical composition;

proof of the rapid emergence of lactoferrin in blood and prolonged action of the pharmaceutical composition;

- proof antitoxic effect created by the claimed invention pharmaceutical compositions.

Example 1

Design rereplacenocase nanoparticles on the basis of the genome of adenovirus human serotype 5 with the insertion of a gene coding for human lactoferrin.

To construct rereplacenocase nanoparticles on the basis of the genome of adenovirus human serotype 5 (the size of 70-80 nm) with the insertion of a gene of human lactoferrin was based on a recombinant plasmid pJM17 (MC Grory W.J., And simple technique for the rescue of early region I mutations into infectious human adenovirus type 5, Virology, No. 163 (2), 1988, s. A simple technique for removal the Oia early region 1 into infectious human adenovirus type 5), with a deletion in region E1 adenoviral genome. All further manipulations on cloning was performed using well known laboratory methods (for example, Sambrook D. and other Methods of genetic engineering. Molecular cloning. M.: Mir, 1984, str-224, 387-420). Cloning was carried out by the method of homologous recombination in cell culture and its essence in the following. Artificially synthesized cDNA gene of human lactoferrin on the chosen restriction enzymes cut sites were periglomerular in the well-known Shuttle plasmid pRCCMV (Invitrogen, San Diego, CA, No. V75020). Next, for the mutual transformation of the obtained plasmid pRcCMV - Lf and vector plasmid pJM17 they were transferrable 293 cells (e.g., CLS, Germany, No. 300192) using the method of calcium phosphate precipitation (Graham F.L. et al., A new technique for the assay of infectivity of human adenovirus 5 DNA, Virology, 1973, No. 52 (2), s-467. - New technique of the method of infection the DNA of human adenovirus 5 serotype). The result was rereplacenocase nanoparticles containing expressing cassette with CMV-promoter, gene of human lactoferrin and a polyadenylation signal. Plaques of recombinant particles were formed on the cell culture in a few days after transfection, they were robbed of a Pasteur pipette, the material obtained was multiplied by the cell line 293 to obtain a titer of 3×1010PC (physical particles/ml (108 IU/ml).

Specified content rereplacenocase nanoparticles and native lactoferrin in the composition of the pharmaceutical composition defined in pharmacokinetics and toxic effects in examples 4 and 5.

To obtain such pharmaceutical compositions cell suspension developed in the previous step containing rereplacenocase nanoparticles in the titer of 3×1010particles/ml, was used for the further expansion of titles rereplacenocase nanoparticles and preparation of finished pharmaceutical compositions containing not less of 2.33×1011FC/ml (which corresponds to the activity of not less than 6.7×108U/ml) and 50-100 μg of native lactoferrin (for example, specified in the patent of Russian Federation №2165769) in 3 ml of composition.

Thus, then, to gain the necessary credits rereplacenocase nanoparticles wave bioreactor with 4500 ml suspension of permissive 293 cell culture were seeded at a cell suspension volume of 500 ml containing rereplacenocase nanoparticles with a titer of 3×1010FC/ml

Cultivated for increasing rereplacenocase nanoparticles inside the cells and achieve their contents 6×1010FC/ml (activity 2×108IU/ml)within approximately 48 hours. Upon reaching the required content of the nanoparticle-cell mass was submitted for clearing, which consisted of several stages:

) Conducted the deposition cell mass by centrifugation. Arriving on clearing the suspension had not less than 1014FC 5 l (estimated using a mass spectrometer, 1 OE (optical unit = 1012FC). Centrifugation was carried out at mode 6000g for 15 min, with liquid adosados was decanted and the remaining solid part containing cells and rereplacenocase nanoparticles were applied for further purification stages.

2) Removing rereplacenocase nanoparticles from the cell culture was performed by cell disruption fourfold primorazhivaniem-thawing. Preparing a buffer solution with a pH of 8.0: 5 ml, 0.075 MNaCl, 1 mMMgCl2, 5% sucrose, 1% Polysorbate 80. Obtained in the previous stage sediment resuspendable in 70 ml of buffer (grade ×71). The solution volume was 80 ml

Freezing was carried out for 2 hours in liquid nitrogen, were thawed in a water bath (37°C), without overheating.

3) To facilitate further removal of cellular genomic DNA was performed additional processing by the nuclease. To this was added benzonase to the concentration in solution of 150 U/ml and placed on a soft stirring with a magnetic stirrer for 3 hours at room temperature (21-23°C).

4) Department rereplacenocase nanoparticles from destroyed cells was carried out by centrifugation at 9000 g for 10 min was Collected supernatant containing rereplacenocase what I nanoparticles.

5) Further purification was performed by ultrafiltration. To this end, the supernatant was diluted with buffer (50 mM TrisHCl pH 7.5, 1M NaCl, 2 mM MgCl2, 5% sucrose, pH 7.5) to a volume of not less than 200 ml, stir with a magnetic stirrer. In the filtering process, the volume of the circulating solution (retentate) constantly brought up to the original (200 ml).

6) Further purification was produced by anion-exchange chromatography.

Retentate was applied on a column (AxiChrom 70/300 400 ml)containing aminoalkenes sorbent Q Sepharose virus licenced. Rereplacenocase nanoparticles were sorbilis on the column, while the impurities did not sorbilis, and was washed with buffer A. After removal of impurities rereplacenocase nanoparticles were desirerable washing buffer B. chromatograph Conditions were as follows: flow 193 ml/min, buffer A (40 mM TrisHCl, 0.27 M NaCl, 2 mM MgCl2, 5% Sucrose, 0.1% Polysorbate 80, pH 7.5), conductivity ~ 28-30 mS/cm; buffer B (40 mM TrisHCl, 0.5M NaCl, 2 mM MgCl2, 5% sucrose, 0.1% Polysorbate 80, pH 7.5) conductivity of ~50 mS/cm. The eluate in a volume of 200 ml was sent to the next stage.

7) Exclusion chromatogaphy

Obtained in the previous stage of the eluate was applied to a column (AxiChrom 100/300 volume 800 ml)containing sorbent Q Sepharose 4 FastFlow. High-molecular substances, not included in the pores of the sorbent, was suirable the first peak (these include rereplacenocase nanoparticles), impurities were suirable on the Le output peak rereplacenocase nanoparticles. Chromatograph conditions were as follows: the flow of 130 ml/min, buffer (10 mMTrisHCl, 75 MNaCl, 1 mMMgCl2,5% sucrose, 0.05% Polysorbate 80, pH 8.0).

To poluchennom eluate (80 ml) was added ethanol to a concentration of 0.5% ethylenediaminetetraacetic acid (EDTA) to a concentration of 100 μm, were sent to the next stage.

8) Normal filtering.

For sterilization of the obtained preparation was carried out filtration through a filter with pore size of 22 μm. The final volume of the drug at this stage was 80 ml and contained rereplacenocase nanoparticles in titer of 1×1012FC/ml was diluted formulating buffer (for example, 10 mMTrisHCl, 75 mMNaCl, 1 mMMgCl2, 5% sucrose, 0.05% Polysorbate 80, 0.5% Ethanol, 100 mm EDTA, pH 8.0) to obtain the content of 2.33×1011FC/ml and sterilized by normal filtering.

Example 2. Obtaining pharmaceutical compositions.

To produce the final composition of the claimed pharmaceutical compositions obtained in the previous stage, the drug was mixed with the concentrate of native human lactoferrin from human milk (Patent RF №2165769), located in the buffer used for the formulation of the drug from rereplacenocase nanoparticles in the previous stage (for example, 10 mm Tris, 75 mm sodium Chloride, 5% sucrose, 0.05% tween-80, 1 mm Magnesium chloride, 0.5% Ethanol, 100 mm EDTA, pH 8.0). Mix the volumes of solution rereplacenocase the camping nanoparticles and concentrate lactoferrin were, in resultate received the specified content rereplacenocase nanoparticles of 2.33×1011FC/ml (which corresponds to the activity of the drug 6.7×108U/ml) and from 50 mg to 100 mg of native lactoferrin in 3 ml of composition.

Example 3.

The stability of the pharmaceutical compositions.

Obtained in example 2 pharmaceutical composition assessed on the stability of the composition.

For this purpose, conducted a visual assessment on close observation of the samples of the drug for 3 minutes. The results of the visual evaluation showed good Miscibility of the components of the drug and no clots.

In table 1. Presents the influence of the components of the pharmaceutical composition on the stability rereplacenocase nanoparticles. The evaluation was made after exposure of pharmaceutical compositions for 0, 30 and 60 minutes with further evaluation of titles rereplacenocase nanoparticles according to standard procedures.

Table 1
The pharmaceutical composition or control substanceThe exposure time, min
03060
titles, FC/m is
The pharmaceutical composition3×1083×1083×108
Culture medium (control substance)3×1083×1083×108

The data in table 1 shows the conservation title rereplacenocase nanoparticles upon exposure of the composition of the pharmaceutical composition from 0 minutes to 1 hour, which corresponds to their preservation in the test substance.

Thus, the obtained results show the stability of the obtained pharmaceutical compositions.

Example 4

Selection of doses of native human lactoferrin and rereplacenocase nanoparticles expressing human lactoferrin.

The possibility of using the pharmaceutical compositions for the treatment of acute toxic conditions was evaluated by its pharmacokinetics during drug laboratory animals (rats) intravenously in a volume containing the dose rereplacenocase nanoparticles expressing human lactoferrin equal to 4.3×1011FC/m2and a dose of native lactoferrin is equal to 10 mg/kg

The evaluation was performed according to the presence and removal of the target protein is lactoferrin human bodies. In the figure depict Alena pharmacokinetic curve, representing the concentration of lactoferrin person in the serum of mice.

Figure 1 shows the pharmacokinetic curve characterizing the concentration of lactoferrin person in the serum of rats after a single intravenous injection of the pharmaceutical composition at a dose rereplacenocase nanoparticles of 4.3×1011FC/m2and native lactoferrin (10 mg/kg

The analysis is shown on figure data showed that the concentration of lactoferrin increases the two peaks. First ascent begins from the moment of introduction of the pharmaceutical composition reaches a peak after 17 minutes with a maximum concentration of Cmax=140 μg/ml and then begins to fall, up to 12 hours after injection (this segment of the curve reflects the dynamics of native lactoferrin composition). However, the fall in the concentration of lactoferrin in whey of blood to 0 does not occur, since the beginning of the 12th hour after administration of the composition is observed the second rise, due to the beginning of the recombinant lactoferrin nerealizirane nanoparticles with a peak at 6.8 days and Cmax=364 µg/ml Then there is a gradual decline in the concentration and lactoferrin completely disappears in the blood by 30 days.

Thus, the concentration of lactoferrin man after a single intravenous injection in the serum of Guinea the rats continuous since the introduction and to 28-30 days with two peaks rise in the concentration of lactoferrin.

If separate single introduction of native lactoferrin in therapeutic dose of 10 mg/kg of the drug from the nanoparticles in formulating the buffer at a dose of 4.3×1013FC/m2as comparative drugs, found that the first peak of the curve is due to native lactoferrin, which disappears by the end of the first day after injection, the second peak corresponds to the time expression recombinante lactoferrin nerealizirane nanoparticles, which starts with only 12 hours after administration and lasts up to 28-30 days. I.e. the joint content of two of these drugs in pharmaceutical compositions allows you to maintain the concentration of lactoferrin human blood since the 17th minute after administration without significant for detoxification therapy fall in the concentration of lactoferrin person.

Thus, evaluation of the pharmacokinetic curve allows us to recommend a pharmaceutical composition for the treatment of not only chronic, but acute toxic conditions, as terapevticheskii action based on detoxifying properties of lactoferrin person starts with 17 minutes after administration and lasts 28-30 days.

Example 5.

The estimation of the detoxifying action of the pharmaceutical composition.

Studied on the model of toxicosis in animals induced by carbon tetrachloride(CCL 4).

The toxicity that occurs with the introduction of the CCl4in the body of a mammal due to the following processes: SSC undergoes metabolic transformation in the membranes of the endoplasmic reticulum of the liver, with the participation of the enzyme cytochrome P-450, which leads to the formation of free radical metabolites (type CCl3from the gap of the molecules CCl4. Due to the intensification of lipid peroxidation lipid complexes of intracellular membranes disrupted the activity of enzymes, some functions of the cell (protein synthesis, metabolism of β-lipoproteins, drug metabolism), there is a degradation of nucleotides, etc. Suggest that the primary site of formation of free radical metabolites are endoplasmic network and microsome assay cells, resulting in decreased activity and degradation of cytochrome P-450, a key enzyme of the microsomal oxidation system. As a result, decreases the rate of metabolism of endogenous and exogenous compounds and weakened the antitoxic function of the liver. The estimation of the detoxifying functions of the liver was performed in hypentelium test, which allows for the duration of narcotic sleep in animals to evaluate the rate of metabolism of thiopental carried out by the cytochrome P-450-dependent monooxygenase system of hepatocytes.

Animals were injected with 75%-n is th oil solution CCl 4subcutaneously (s/C) a single dose of 2 ml/kg at the same time experimental mice were administered pharmaceutical composition once intravenously at a dose of 4.3×1011FC/m2and 10 mg/kg of native human lactoferrin. Control groups once intravenously injected drug, containing only the nanoparticles expressing human lactoferrin at a dose of 4.3×1011FC/m2or 0.9% sodium chloride. Thiopental was administered on the 6th day after the introduction of the CCl4intraperitoneally (b) a single dose of 55 mg/kg and recorded sleep duration in experimental animals, as a criterion for assessing the degree of hepatic toxicity.

Figure 2 presents data on the duration thiopental sleep in animals inoculated CCl4. The bars indicate the duration of sleep in groups of animals, which were introduced:

1 - CCl4;

2 - 0.9% solution of sodium chloride;

3 - native human lactoferrin;

4 is a drug containing only rereplacenocase nanoparticles with the insertion of a gene lactoferrin;

5 is a pharmaceutical composition.

In the figure, the data shows that the introduction of the CCl4increases the duration of sleep of animals in comparison with the duration of sleep in control animals treated with physiologic solution (41±15 min and 5±2 min, respectively)that the witness is there about the decrease in the rate of metabolism of thiopental in the liver and, accordingly, about the weakening of the antitoxic function of the liver.

Accordingly, the presented results indicate that the duration thiopental sleep in the experimental group (20±6 min)was significantly lower compared with the control group of mice (41±15 min), treated with CCl4but have not received the pharmaceutical composition, which means that there detoxifying properties of the claimed pharmaceutical compositions. In addition, sleep in animals that received only native human lactoferrin or preparation containing rereplacenocase nanoparticles with insert lactoferrin gene was reduced in comparison with a group of CCl4to 23±8 min, but it was a little longer in comparison with an experienced group, indicating the presence of the best detoxifying properties of the claimed composition.

Thus, a single intravenous administration of the pharmaceutical composition during the onset of acute toxicity caused by the introduction of the CCl4significant detoxicate neck effect on the body, which is stronger than the separate introduction in comparable quantities of drugs native lactoferrin and rereplacenocase nanoparticles expressing lactoferrin.

Further, in clinical examples, the dose received during the pre-clinical studies and measured on m2PAE is rnost body, have been translated into humans, because they are equivalent (average surface area of the body is equal to 1,62 m2). (Gabriel RU, Manual on experimental pre-clinical study of new pharmacological substances, 2000, p.98), (Guidance for Industry. Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER), Pharmacology and Toxicology, USA, 2005, p.7, 19. - Guidance for industry. Evaluation of the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers).

Example 6

Patient P. Arrived with the clinical picture of acute toxic gastroenteritis. The main symptoms are salivation, vomiting, diarrhea, cramping abdominal pain for several hours. Was carried out gastric lavage, administration of diuretics, intravenous infusion of saline, the treatment was supplemented by the introduction of the claimed pharmaceutical compositions once intravenously in a volume of 3 ml, which corresponds to the introduction rereplacenocase nanoparticles expressing lactoferrin equal dose of 7×1011FC and native lactoferrin at a dose of 50 mg per person. Within hours after the emergency has passed salivation and vomiting, decreased pain, and diarrhea. The patient's condition has improved. By the end of the first day is to assist the clinical symptoms were completely gone, toxicosis was docked.

Example 7.

Patient C. Acute poisoning with ethyl alcohol. The main clinical symptoms - cold clammy skin, flushing of the face and conjunctiva, lower body temperature, vomiting, involuntary urination and feces, eyes narrowed, and with an increase of respiratory distress dilate, breathing slow, the pulse frequent, weak. On the background of standard infusion therapy was conducted removing toxic shock introduction 3 ml of the claimed pharmaceutical compositions once intravenously in a volume of 3 ml, which corresponds to the introduction rereplacenocase nanoparticles expressing lactoferrin equal dose of 7×1011FC and native lactoferrin at a dose of 100 mg per person. The patient's condition after the implementation of an antitoxic therapy has been significantly improved, breathing and heartbeat returned to normal, the surface of the skin and mucous membranes, and eyes. By the end of the first day of treatment, the clinical symptoms disappeared completely, the General condition of the patient improved significantly.

Example 8.

The patient s Diagnosis: cancer of the colon, post-surgical and chemotherapeutic treatment. In the postoperative period the patient developed toxic hepatitis. After injection of 3 ml of the claimed pharmaceutical compositions once intravenously in a volume of 3 ml, which corresponds to the introduction of the of acoustic expressing lactoferrin equal dose of 7×10 11FC and native lactoferrin at a dose of 50 mg per person. Revealed: the decrease of total and direct bilirubin in serum 105/80 µmol/l → 8,4/3.9 µmol/l Toxic hepatitis was arrested.

Use in the pharmaceutical and clinical practice of the claimed pharmaceutical compositions allows to achieve several technical, medical and economic results:

- the claimed pharmaceutical composition is biocompatible with the human body and therapeutically efficient;

pharmaceutical composition suitable for use as entered once and then, starting in the 17th minute after injection, and within within 28-30 days long produces in the human body the human lactoferrin, creating blood concentrations, ten times the normal level, and desired to achieve a stable therapeutic effect;

- the use of the pharmaceutical composition is economically justified, since a single injection of a medicinal product provides a fast and prolonged therapeutic effect;

- the reduction of labor costs medical staff, medical instruments, and thus the complexity and cost of treatment, as native lactoferrin requires frequent introduction, this may be avoided by the introduction of rereplacenocase nanocat the CI which produce large quantities of lactoferrin directly in humans after a single injection;

- reducing the need for native lactoferrin, as donor human milk is scarce.

These examples show that the resulting pharmaceutical composition which allows after a single intravenous injection to obtain an effective anti-toxic effect since the 17th minute after its introduction, to 28-30 days after injection, which allows for therapy of various toxic conditions, particularly acute. Thus, the goal of the project is executed.

1. Pharmaceutical composition for the treatment of acute toxic state, containing the protein human lactoferrin, characterized in that it further comprises rereplacenocase nanoparticles with the insertion of a gene of human lactoferrin and formulating the buffer in the following ratio of components per dose:
lactoferrin man from 50 to 100 mg;
rereplacenocase nanoparticles - 7·1011physical particle;
formulating buffer - rest, Jr.

2. The pharmaceutical composition according to claim 1, characterized in that the dose is 3 ml

3. The pharmaceutical composition according to claim 1, characterized in that the human lactoferrin - lactoferrin donor breast milk.

4. Pharmaceutical is Kai composition according to claim 1, wherein the lactoferrin human - any human lactoferrin.



 

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2 cl, 3 ex

FIELD: measurement equipment.

SUBSTANCE: method is proposed to manufacture a vacuum sensor with a nanostructure, consisting in the fact that a thin-film semiconductor resistor is formed in the form of a meshy nanostructure (SiO2)50%(SnO2)50% by application of a sol of orthosilicic acid containing a stannum hydroxide, onto a silicon substrate with the help of a centrifuge and subsequent baking. The sol is prepared in two stages, at the first stage tetraetoxysilane and ethyl alcohol are mixed, then at the second stage distilled water is added to the produced solution, as well as hydrochloric acid (HCl) and stannum chloride dihydrate (SnCl2·2H2O). The vacuum sensor with the nanostructure made according to the proposed method comprises the body, the heterogeneous structure installed in it from thin films of materials, formed on the substrate of a semiconductor, the thin-film semiconductor resistor and contact sites to it, formed in the heterogenerous structure, leads of the body and contact conductors, which connect contact sites with body leads.

EFFECT: increased sensitivity of a vacuum sensor.

5 cl, 4 dwg

FIELD: metallurgy.

SUBSTANCE: metal strip contains a coating from carbon nanotubes and/or fullerenes soaked with metal chosen from the group consisting of Sn, Ni, Ag, Au, Pd, Cu, W or their alloys. Method for obtaining metal strip with the coating from carbon nanotubes and/or fullerenes and metal involves the following stages: a) application of diffusion barrier layer from transition metal Mo, Co, Fe/Ni, Cr, Ti, W or Ce onto a metal strip, b) application of a nucleation layer from metal salt containing metal chosen from the group Fe, the 9-th or the 10-th subgroup of the periodic table onto the diffusion barrier layer, c) introduction after stages a) and b) of treated metal strip to hydrocarbon atmosphere containing organic gaseous compounds, d) formation of carbon nanotubes and/or fullerenes on metal strip at temperature of 200°C to 1500°C, e) soaking of carbon nanotubes and/or fullerenes with metal chosen from the group containing Sn, Ni, Ag, Au, Pd, Cu, W or their alloys.

EFFECT: obtained metal strip with the coating has improved friction coefficient, increased transition resistance of a contact, increased resistance to friction corrosion, improved resistance to abrasion and increased ability to be deformed.

26 cl

FIELD: chemistry.

SUBSTANCE: invention can be used to make damping elements, shock absorbers, friction pairs and wear-resistant components of micromechanisms. Starting carbon material is placed in a working volume; nitrogen is pumped and removed until complete displacement of air. At the first step, nitrogen is pumped to working pressure of 220-250 MPa, heated to 1670-2020 K and held for not less than 1 minute. Temperature is then lowered to room temperature and pressure is lowered to atmospheric pressure. At the second step, the obtained carbon-nitrogen material is treated in a high-pressure apparatus at pressure of 7-15 GPa and temperature of 1620-1770 K for not less than 1 minute. Temperature is then lowered to room temperature and pressure is lowered to atmospheric pressure. The starting carbon material used is fullerites, for example C60 and/or C70, which are first ground to particle size smaller than 1 mcm in order to increase content of nitrogen in the finished material.

EFFECT: invention enables to obtain a compact carbon-nitrogen material with a bulbous structure, which contains 2,2-5,0 at % nitrogen, having Young's modulus of 43-67 GPa, microhardness of 5,4-8,2 GPa and hyperelastic recovery parameter of 92-97%.

3 cl, 5 dwg, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a method for preparing stabiliser-coated nanocrystalline cerium dioxide characterised by antioxidant activity. The method involves preparing an aqueous solution of cerium salt and a stabiliser representing maltodextrin with a molar ratio of cerium to the stabiliser 1 to 1-4. Then, the prepared aqueous solution is added with drops of hydrous ammonia with stirring, and pH of the prepared solution is gradually increased to 7-8, maintained for 1-4 hours, the prepared colloidal solution of hydrous cerium nanoparticles is added with hydrous ammonia, and pH is increased to 11-12 and maintained for 1-10 hours to form a colloidal solution of cerium dioxide. Thereafter, an alcohol or ketone excess is removed and brought to the boiling point, while the formed precipitate of the non-aggregated nanoparticles of stabiliser-coated cerium dioxide, separated by decantation or filtration, washed 1-4 times in alcohol or ketone, and dried at temperature 50-80°C to constant weight. The prepared powder of the non-aggregated nanoparticles of stabiliser-coated cerium dioxide is re-dispersed in a polar solvent to form aggregation resistant sol.

EFFECT: invention provides preparing stabilised nanocrystalline cerium dioxide with a hydrodynamic diameter of 6-10 nm.

3 cl, 7 dwg, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and pharmacy and can be applied in production and application of solutions for intravenous introduction in treatment of conditions, associated with endogenous intoxication. Disintoxication solution contains components in the following ratio (wt %): sodium hypochlorite 0.04-0.08, sodium chloride 0.50-1.00, aminoethane sulfonic acid 0.02-1.20.

EFFECT: novel infusion solution is claimed.

4 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to a novel chemically stable antioxidant compound which contains a lipophilic cationic moiety linked by a linking moiety to an antioxidant molecule and an anionic component for said cationic moiety, where the antioxidant compound is a mitoquinone, selected from: 10-(6'-ubiquinone)propyltriphenylphosphonium, 10-(6'-ubiquinonyl)pentyltriphenylphosphonium, 10-(6'-ubiquinonyl)decyltriphenylphosphonium, and 10-(6'-ubiquinonyl)pentadecyltriphenylphosphonium, having general formula I: or quinol form thereof, where R1, R2 and R3 denote CH3, the C atom in (C)n is saturated and n equals 3, 5, 10 or 15, and Z denotes an anionic component which is selected from a group consisting of methanesulphonate and ethanesulphonate. The invention also relates to a pharmaceutical composition for reducing oxidative stress in a cell, containing said compound and optionally containing β-cyclodextrin, a method of reducing oxidative stress in a cell and a method of producing an antioxidant compound.

EFFECT: improved properties of compounds.

25 cl, 31 dwg, 13 tbl, 11 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a novel compound - 5-hydroxy-6-methyl-1-(thietanyl-3)pyrimidine-2,4(1H,3H)-dione of formula , which inhibits generation of active forms of oxygen and has antioxidant activity.

EFFECT: improved properties of compounds.

2 cl, 1 dwg, 1 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to a novel compound - 6-methyl-1-(thietanyl-3)uracil of formula 1 , which stimulates the protective activity of phagocytes.

EFFECT: obtaining compounds which stimulate the protective activity of phagocytes.

2 cl, 1 dwg, 1 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to organic chemistry and medicine and specifically to a novel compound - 6-(thietanyl-3)aminopyrimidine-2,4(1H,3H)-dione of formula (1), which inhibits lipid peroxidation.

EFFECT: obtaining a compound which inhibits lipid peroxidation.

2 cl, 1 dwg, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a method for preparing stabiliser-coated nanocrystalline cerium dioxide characterised by antioxidant activity. The method involves preparing an aqueous solution of cerium salt and a stabiliser representing maltodextrin with a molar ratio of cerium to the stabiliser 1 to 1-4. Then, the prepared aqueous solution is added with drops of hydrous ammonia with stirring, and pH of the prepared solution is gradually increased to 7-8, maintained for 1-4 hours, the prepared colloidal solution of hydrous cerium nanoparticles is added with hydrous ammonia, and pH is increased to 11-12 and maintained for 1-10 hours to form a colloidal solution of cerium dioxide. Thereafter, an alcohol or ketone excess is removed and brought to the boiling point, while the formed precipitate of the non-aggregated nanoparticles of stabiliser-coated cerium dioxide, separated by decantation or filtration, washed 1-4 times in alcohol or ketone, and dried at temperature 50-80°C to constant weight. The prepared powder of the non-aggregated nanoparticles of stabiliser-coated cerium dioxide is re-dispersed in a polar solvent to form aggregation resistant sol.

EFFECT: invention provides preparing stabilised nanocrystalline cerium dioxide with a hydrodynamic diameter of 6-10 nm.

3 cl, 7 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely to radiation biology, and may be used to treat a cadmium radiation injury and to produce a preparation for the cadmium radiation injury. Treating the cadmium radiation injury is ensured by the single subcutaneous administration of a composition consisting of anti-radiation therapeutic globulin and a suspension-forming fraction of bentonite, in a dose of 20-25 mg/kg of body weight that is followed by three more administrations 24, 48, 96 hours later if observing chronic heavy metal administration. The preparation is produced by dissolving the suspension-forming fraction of bentonite in anti-radiation therapeutic globulin in ratio 0.3:99.7 respectively while constantly stirred, and a solid concentration is reduced to 10%; the preparation is sterilised by filtration and then bottled 250-300 cm3 each to be closed tightly and kept at temperature 4-6°C.

EFFECT: method enables a consistent process of the preparation with enhanced therapeutic and decorporating effect ensured by the targeted delivery as a single agent with simplifying the use conditions.

3 cl, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely an agent possessing anti-inflammatory action. The agent possessing anti-inflammatory action contains as active ingredient in the form of a complex prepared of a coelomic fluid of sea urchins containing peptides and amino acids at a certain ratio of the ingredients.

EFFECT: agent possesses a manifested anti-inflammatory effect.

2 cl, 2 tbl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is the use of L-carnosine for making a nanopreparation having antihypoxic and antioxidant activity combined with a combination of substances selected from the group of phospholipids, non-polar lipids in the following ratio, wt %: L-carnosine - 1.1-1.2, non-polar lipids such as triglycerides, cholesterol, free fatty acids, DL-α-Tocopherol - 1.2-2.5, phospholipids such as phosphatidylcholine, phosphatidylethanolamine, lysophosphatidylcholine, lysophosphatidylethanolamine, sphingomyelin - 95.3-96.3 for preparing a drug having antihypoxic and antioxidant activity. The drug can be presented in the form of liposomes containing L-carnosine.

EFFECT: invention provides higher stability of L-carnosine and its lifetime up to three days with underlying higher effectiveness in small doses, as well as to improve the cerebral ischemia tolerance, the recovery after acute hypoxia and to increase the antioxidant status of the brain tissue.

3 cl, 4 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to a method for preparing a carbon sorbent with the antibacterial properties, and to the carbon sorbent with the antibacterial properties prepared by this method. The declared method involves impregnation of the carbon hemosorbent granules in an initiator solution in N-vinylpyrrolidone at pH 7.0-7.5 and a residual pressure of 15-20 mm Hg. The hemosorbent : initiator solution in N-vinylpyrrolidone ratio is 1:1.4-2.0. Then the temperature is raised to 65-75°C, kept at that temperature for 0.5-8 hours in an inert medium and washed in water from the residual monomer at room temperature.

EFFECT: carbon sorbent with the antibacterial properties prepared by the specified method represents the round granules, contains polyvinylpyrrolidone in an amount of 4,5-5,5% and is characterised by a specific surface adsorption of less than 50 m2/g and total pore volume of less than 0,30 cm3/g.

2 cl, 2 tbl, 1 dwg, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to introducing dairy fat or an analogue thereof, optionally with at least one additional therapeutic factor, preferentially with lactoferrin or lactoferrin containing a metal ion, preferentially lactoferrin containing iron, preferentially bovine lactoferrin containing iron or a functional version thereof containing metal ions or a functional fragment to inhibit the tumour formation or growth, to maintain or improve one or more parameters, such as leukocyte count, erythrocyte count, or myelocyte count, to reduce the manifestations of cachexia, mucositis and anemia, to stimulate the immune system and to treat or prevent a malignancy and malignant symptoms, and side effects of treating the malignant tumour. Methods and therapeutic use according to the invention may be implemented by the use of a diet (in the form of food products or food additives), nutrient or pharmaceutical composition. There are also presented compositions applicable in the methods according to the invention.

EFFECT: group of inventions provides the higher clinical effectiveness or prevention of the malignant tumour or its symptoms.

39 cl, 7 tbl, 21 ex

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