Chimeric peptide for epithelial and mesenchymal malignant neoplasms treatment
SUBSTANCE: invention relates to medicines, particularly to the use of chimeric peptide VP-22_p16INK4a for epithelial and mesenchymal malignant neoplasms treatment. The claimed chimeric peptide VP-22_p16INK4a contains two amino acid sequences. The first sequence comprises inhibitor of cycline kinases as active fragment p16INK4a as therapeutic agent and the second sequence comprises peptide VP22 of herpes simplex virus as carrier agent to deliver cycline kinase inhibitor into target cells.
EFFECT: enlarging the application range of medicine.
The invention relates to medicine, namely to chimeric peptides containing transport (chimeric, in the world literature there is also the term "cell penetrating peptides" - RAF) part and the functional part, intended for the treatment of epithelial and mesenchymal malignant neoplasms (i.e. malignant neoplasms of non-lymphoid origin).
The creation of chimeric peptides with cytostatic and cytotoxic activity, is a promising direction of development of modern therapy of malignant tumors of non-lymphoid origin.
From the description of the patent RU 2297241 C2 invention Chimeric protein for the treatment of malignant lymphomas" (published 20.04.2007) it is known that one of the areas of treatment of neoplastic diseases is the use of inhibitors ticinovic kinases, for example protein family INK4a affecting the cell cycle. It is also known that it is desirable that the molecular constructs were delivered into the cell "address" and hit it on a specific intracellular signal or function. From the above document is also known that depending on the structure of different peptides may accumulate in different compartments of the cell, which gave a theoretical possibility to construct after which outlinesthe with a target delivery to different organelles of the cell.
Chimeric peptide VP-22_p16INK4a the patent RU 2297241 C2 has a higher biological effect due to the improved transport properties of the agent and is intended for the treatment of malignant lymphomas, however, is known of the possibility of its use for the treatment of epithelial and mesenchymal malignant tumors.
The present invention is the expansion of the use of the chimeric peptide VP-22_p16INK4a and the application of this peptide for the treatment of epithelial and mesenchymal malignant tumors.
The technical result of the invention is medical-biological effect, objectively manifested in the use of this chimeric peptide consists of the solution of the stated problem.
The technical result is achieved due to the use for the treatment of non-lymphoid neoplasms of origin chimeric peptide VP-22_p16INK4a containing two amino acid sequences, the first of which includes the inhibitor ticinovic kinases in the form of an active fragment of p16INK4a (amino acid residues 84-103 or 84-106) as a therapeutic agent, and the second includes the VP22 peptide (amino acid residues 140-301) of herpes simplex virus as a transport agent for transfer inhibitor ticinovic kinases inside the target cells.
Himem the th peptide VP-22_p16INK4a contains the following amino acid sequence: D-A-A-T-A-T-R-G-R-S-A-A-S-R-P-T-E-R-P-R-A-P-A-R-S-A-S-R-P-R-R-P-V-E- D-A-A-R-E-G-F-L-D-T-L-V-V-L-H-R-A-G-A-R(hereinafter referred to SEQ ID NO. 1, underlined sequence of the P16).
Control and synchronization of events in the process of cell division is a large complex molecules, one of the key participants in this process are cycline, activating the so-called cyclin dependent kinases (CDK). During the cell cycle is a sequential activation of transcription of specific cyclina with the subsequent formation of the active complex of the cyclin-CDK. Stop cell division in the control (restriction) points (G1, S, G2) is the inhibition of the corresponding zaklinivalo complex through specific proteins. A family of proteins, inhibiting cyclin D (controlling the transition of G1-S) refers to the peptides INK4a.
The most studied of this group p16INK4a peptide. This peptide inhibits the activity of CDK4 and CDK6, included in the complex with cyclin D, which inhibits pRb phosphorylation and the release of E2F, and leads to cell cycle arrest at the border of the G1-S transition. Structural and functional studies of this protein have revealed active fragment of p16INK4a (amino acids 84-103 or 84-106), which is responsible for the inhibition of CDK4 and CDK6, included in the complex with cyclin D.
The p16INK4a peptide plays an important role in the process of differentiation and aging of cells, in addition, this peptide is not functional is h or absent in many tumor tissues. For malignant neoplasms of non-lymphoid origin characterized by impaired function of p16INK4a peptide and overexpression of Zilina D. Thus, in malignant tumors of non-lymphoid origin there is impaired control restriction point R1 (G1-S transition).
As mentioned in the description of the patent RU 2297241 C2, the ability to use natural inhibitors ticinovic kinases to control proliferation occurred after the discovery of the ability of some proteins to the TRANS-activation. That is, proteins synthesized in the same cell, could go out and penetrate into a different cell, lead to the activation of certain genes. The study of the structure of these proteins allowed us to identify short (10 to 30 amino acids) amino acid sequences responsible for intracellular transport. It was shown that the addition of such a sequence in the structure of an arbitrary protein confers properties of intracellular and intranuclear internalization. Currently, there are about 30 known sequences of such peptides.
Constructed chimeric molecule peptide comprising VP22 protein fragments and a fragment of the protein p16INK4a inhibitor ticinovic kinase, previously used exclusively for the treatment of malignant lymphomas. Currently, however, there are experimental the data validate the use of the chimeric peptide VP-22_p16INK4a and for the treatment of epithelial and mesenchymal malignant tumors.
The possibility of carrying out the invention with the implementation of the specified destination confirm these results.
Materials and methods. Examples
Working with cell cultures
Cells were cultured at 37°C in 5% CO2 atmosphere in DMEM (Paneco, Russia)containing 10% fetal bovine serum (FSB) (Paneco, Russia), 10 μg/ml of antibiotic-antimycotics gentamicin (Paneco, Russia) and 2 mm L-glutamine (Paneco, Russia).
Chimeric peptide and anticancer drugs were dissolved in culture medium and added to cells at changing environment. The effect was evaluated after 24 hours of incubation.
Short-term culture of human cancers
To assess the effectiveness of the cytotoxic effect of chimeric peptide VP-22_p16INK4a on tumor person used short-term cultures derived from operational material breast cancer, cancer of the body of the stomach, cancer of the cervix and kidney cancer. It was also evaluated the effect of chimeric peptide in short-term cultures of skin, unchanged mucosa of the stomach, unmodified endometrium and unmodified kidneys. Short-term culture was obtained by mechanical disaggregation of the operational material, the cells were incubated 24 hours at 37°C in a 5% atmosphere of CO2in medium RPMI (Paneco, Russia)containing 5% fetal bovine serum (FSB) (Paneco, Russia) and d is I the growth components. Next was replacing the medium on the medium containing chimeric peptide VP-22_p16INK4a at a concentration of 40 μm. The effect was evaluated after 24 and 48 hours of incubation with the peptide.
When working in vitro effects on the cells of chimeric peptides was evaluated using the method of flow cytofluorimetry (flow cytometer DAKO Galaxy). We studied the level of apoptosis, the number of dead cells by staining AnnexinV-PI (propedy iodide) and quantitative distribution of cells in cell cycle phases in coloration PI.
Coloring on AnnexinV. Used set of AnnexinV KIT (Caltag laboratories LI 2004). Cells were washed from the environment in cold phosphate-buffered saline (PBS), resuspendable in 1 × Binding Buffer at a concentration of 106 cells per ml were Collected and 100 μl of the resulting suspension was added to 5 μl of AnnexinV-FITC and 10 μl PI. Incubated in the dark at room temperature for 15 minutes. Added 400 μl of 1 × Binding Buffer and analyzed on a flow cytometer.
Colour PI. Cells were washed from the serum twice by centrifugation in 1 ml PBS (4 minutes at 2000 rpm). Removed supernatant, was added 300 μl of PBS was added dropwise to 700 μl of ice-cold 70% ethanol and kept at -20°C overnight (fixation). Fixed cell samples were besieged by centrifugation (4 min at 2000 rpm), washed with 1 ml PBS and repeat the centrifugation. To the precipitated cells were added 100 μl of PBS and 20 μl of RNA is PS (1 mg/ml) and incubated for 30 minutes at 37°C. Further to the suspension was added to 30 μl of PI and incubated for 40 minutes at room temperature in the dark. After the incubation was finished, the volume of sample to 1 ml with PBS.
In the study of chimeric peptides in vivo antitumor effect was evaluated in Nude mice (nude) with inoculated tumor human cell lines A (lung cancer) and HCT-116 (colon cancer person).
To study the effects on human tumors used method of short-term cultures.
Investigation of the effect of chimeric peptide in short-term culture of breast cancer, stomach cancer, kidney cancer. It is shown that for this model the most sensitive option tumor is a cancer of the kidney.
Experiments in vivo
In Nude mice (Nude) studied the antitumor activity of chimeric chimeric peptide VP-22_p16INK4a. Mice were subcutaneously inoculated tumor human cell culture lines A (lung cancer) and HCT-116 (colon cancer) in an amount of about 1 million cells per mouse. Introduction peptide was started after detection of the tumor site (3-4 mm) on 4-6 day after inoculation of the tumor. Introduced 100 (first group) or 200 μg (second group) of the peptide through the day. For both tumor lines received a 50% inhibition of tumor growth. Differences in the effect for doses of 100 and 200 µg not found.
The results of issledovaniye malignant neoplasms of non-lymphoid origin using chimeric peptide VP-22_p16INK4a explained by the following drawings:
Figure 1: the level of apoptosis in short-term cultures unchanged renal tissue (normal) and renal carcinoma (cancer) when incubated with the chimeric peptide VP-22_p16INK4a at a concentration of 40 μm for 24 hours. Mean values from 4 experiments. The number of apoptotic particles was determined as positive for AnnexinV with dual color AnnexinV-PI;
Figure 2: cytotoxic effect of chimeric peptide VP-22_p16INK4a short-term culture unchanged renal tissue (normal) and renal cancer (cancer). The average number of particles, positive for AnnexinV and particles, including PI, double color AnnexinV-PI on four experiments. Incubation with chimeric peptide for 24 hours;
Figure 3: the level of apoptosis in short-term cultures unchanged tissue of the gastric mucosa (normal) and gastric cancer (cancer) when incubated with the chimeric peptide VP-22_p16INK4a at a concentration of 40 μm for 24 hours. Mean values from 4 experiments. The number of apoptotic particles was determined as positive for AnnexinV with dual color AnnexinV-PI;
Figure 4: the cytotoxic effect of chimeric peptide VP-22_p16INK4a short-term culture unmodified tissue of the gastric mucosa (normal) and gastric cancer (cancer). The average number of particles, positive for AnnexinV and particles, including PI, double color AnnexinV-PI on four experiments. Incubation with himem the m peptide for 24 hours;
Figure 5: level of apoptosis in short-term cultures unmodified body tissue of the uterus (the norm) and cervical cancer (cancer) when incubated with the chimeric peptide VP-22_p16INK4a at a concentration of 40 μm for 24 hours. Mean values from 4 experiments. The number of apoptotic particles was determined as positive for AnnexinV with dual color AnnexinV-PI;
6: the cytotoxic effect of chimeric peptide VP-22_p16INK4a short-term culture unmodified body tissue of the uterus (the norm) and cervical cancer (cancer). The average number of particles, positive for AnnexinV and particles, including PI, double color AnnexinV-PI on four experiments. Incubation with chimeric peptide for 24 hours;
Fig.7: the level of apoptosis in short-term cultures unchanged tissue skin breast (normal) and breast cancer (cancer) when incubated with the chimeric peptide VP-22_p16INK4a at a concentration of 40 μm for 24 hours. Mean values from 4 experiments. The number of apoptotic particles was determined as positive for AnnexinV with dual color AnnexinV-PI;
Fig: the cytotoxic effect of chimeric peptide VP-22_p16INK4a short-term culture unmodified tissue skin breast (normal) and breast cancer (cancer). The average number of particles, positive for AnnexinV and particles, including PI, double color AnnexinV-PI on four set of experiment is. Incubation with chimeric peptide for 24 hours;
Figure 9: change in the volume of tumor in nude mice. Mice were inoculated subcutaneously cells HCT-116 (colorectal cancer) is the number of 1 million Chimeric peptide VP-22_p16INK4a dose of 0.1 mg was administered to the experimental group (experience) in the area of the tumor site, the control group (control) in the area of the tumor site was injected with saline. The first injection was made on the 6th day after inoculation of the cells HCT-116.
The results of the experiments shown in figures 1 to 9, which shows an increase in the level of apoptosis and necrosis in malignant tumors of non-lymphoid origin when using chimeric peptide VP-22_p16INK4a confirm pronounced cytostatic and cytotoxic effect against non-lymphoid tumor Genesis in both in vitro and in animal models.
The described experiments on the intratumoral injection of peptides in subcutaneous variant inoculation of the tumor were the first in the world. In the literature are not described similar experiments.
The results on short-term models of human tumors obtained for the first time in the world. First it is shown that the peptide with the structure of VP-22_p16INK4a can be applied for the treatment of malignant tumors of non-lymphoid origin.
The use of chimeric peptide VP-22_p16INK4a containing the amino acid sequence of SEQID NO.1, for the treatment of epithelial and mesenchymal malignant tumors.
SUBSTANCE: there is offered method for identification and/or verification of inhibitors of receptor tyrosine kinases that involves application of a new test system which represents a yeast host cell containing an expression vector including a nucleic acid sequence that encodes fused protein essentially consisting of a complete cytoplasmic part of analysed receptor tyrosine kinase and the dimerisation domain and, if necessary, in addition including anchoring sequence for fused protein in a membrane wherein expression of fused protein conduces to termination of cell proliferation. The method provides production of specified host cells being in contact with a candidate compound and identification of inhibitors of tested tyrosine kinase activity as a result of cultivation on the assumption that inhibition of tyrosine kinase activity with a candidate compound causes restoration of proliferation process.
EFFECT: prospected application of the invention is related to development of selective therapeutic, including anticancer, agents.
13 cl, 5 dwg, 2 ex
SUBSTANCE: proposed is a recombinant single-strand trispecific antibody for treating tumours which express CEA. The said antibody consists of a series of three antibody fragments: anti-CEA-scFv, anti-CD3-scFv and VH CD28-antibody, linked by two intermediate linkers (intermediate linker Fc and intermediate linker HSA). If necessary, a c-myc-mark or (His)6-mark can be added at the C-end. Described is DNA, which codes the antibody, expression vector based on it and E.coli cell, containing the vector.
EFFECT: use of the invention is more beneficial in clinical use compared to bispecific antibodies and known trispecific antibodies, makes easier clearing and expression of an antibody, which can further be used in treating CEA-mediated tumours.
10 cl, 21 dwg, 11 ex
SUBSTANCE: hybrid protein - human insulin precursor consists of N-end fragment of human gamma-interferon connected through peptide linker with amino acid sequence of human proinsulin. Recombinant human insulin is obtained by cultivation of Escherichia coli JM109/pHINS11 strain-producer, carrying plasmid pHINS11, isolation of inclusion bodies and their dissolving in buffer which contains urea and dithiotreitole. Then hybrid protein re-naturation, sedimentation of admixture compounds, purification of re-naturated hybrid protein by ion-exchanging chromatography, combined fermentative hydrolysis of hybrid protein with tripsin and carbopeptidase B are carried out. At the last stage insulin purification with cation-exchanging chromatography and method of highly efficient reverse phase liquid chromatography are carried out.
EFFECT: simplification of obtaining highly purified recombinant human insulin and increase of its output.
6 cl, 1 dwg, 4 tbl, 5 ex
FIELD: chemistry, biochemistry.
SUBSTANCE: current invention relates to the field of biotechnology and immunology. Proposed is an antibody, specific to the human ED-B. Antibody specified is a molecule in the form of either dimerizated mini-immunoglobulin or IgG1, whose variable region comes from the antibody L19. In case the mini-immunoglobulin variable region L19 is merged with εS2-CH4, then as in the case IgG1, the variable region L19 is merged with the constant domain of IgG1. Conjugates of antibodies with radioisotopes have been discovered. Described is the coding nucleic acid, carrying its host cell, capable of producing antibodies, and method of obtaining antibodies from cells. Discovered is a method of determining the degree of bonding of antibodies, also compositions based on antibodies. Described is the use of antibodies for preparing medicine for treating either damage related to angiogenesis, or for treating tumours. Utilisation of the invention provides antibodies, which possess high accumulating capacity to tumours, improved capability to bonding with radioactive labels and unexpectedly retains immunoreactivity in the plasma, in comparison to scFv L19. Antibody specified can be used in diagnostics and treatment of tumours.
EFFECT: obtaining antibodies which can be used in diagnostics and treatment of tumours.
22 cl, 13 dwg, 8 tbl
FIELD: chemistry; biochemistry.
SUBSTANCE: present invention pertains to biotechnology and can be used in biomedicine for producing hyaluronan. Proposal is given of a method of producing hyaluronan, involving culturing Bacillus host cells in conditions, suitable for obtaining hyaluronan, and extraction of the target product from the culture medium. The Bacillus host cell contains a genetic structure, comprising a promoter, functionally active in the given cell, and encoding a region, consisting of a nucleotide sequence, encoding streptococcal hyaluronansynthase (hasA); sequence encoding UDP-glucose-6-dehydrogenase Bacillus (tuaD) or a similar enzyme of streptococcal origin (hasB), and a sequence, encoding bacterial or streptococcal UDP-glucose pyrophosphorylase (gtaB and hasC respectively). Use of the invented method provides for production of considerable quantities of hyaluronan with good examined, nonpathogenic cellular system.
EFFECT: obtaining considerable of hyaluronan with good examined, nonpathogenic cellular system.
15 cl, 45 dwg, 2 tbl, 20 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: present invention pertains to genetic engineering, more specifically to chimeric polypeptides, containing an antagonist of growth hormone receptor. The invention can be used in medicine. The binding domain of the growth hormone is modified by substituting glycine amino acid residue in position 120 and is further modified in site 1, where at least one amino acid residue is substituted, which increases affinity of the growth hormone to its binding domain on the growth hormone receptor. The amino acid residue is then conjugated with the ligand-binding domain of the growth hormone receptor, through a peptide linker.
EFFECT: obtaining a highly effective antagonist of the growth hormone receptor with longer half-life, reduced immunogenesity and nontoxicity, compared to known mutant forms.
35 cl, 16 dwg, 1 tbl
SUBSTANCE: invention claims compositions which can include one or several mammary gland tumour proteins, their immunogenic parts or polynucleotides encoding such parts. Alternatively the therapeutic composition can include antigen-presenting cell expressing mammary gland tumour protein, or T-cell specific to cells expressing such protein. These compositions can be applied in prevention and treatment of such diseases as mammary gland cancer. Invention also claims diagnostic methods based on determination of mammary gland tumour protein or mRNA encoding such protein in sample.
EFFECT: use of peptides obtained from protein expressed from mammary gland by tumour in diagnostics and therapy of mammary gland cancer.
37 cl, 6 ex, 1 dwg
FIELD: medicine; pharmacology.
SUBSTANCE: immunogenic hybrid polypeptide includes mimetic peptide of V-cellular epitope of apolypoprotein B-100 in which C-end of mimetic peptide is merged with N-end of T-helper epitope. Amino acid sequences of polypeptide variants are presented in description. Described is method of specified polypeptide production providing application of host cell transformed with recombinant express vector including gene coding specified polypeptide. Besides, invention concerns vaccine composition including specified immunogenic hybrid polypeptide for obesity prevention or treatment, recombinant express vector and host cell.
EFFECT: excellent anti-obesity activity without induction of immune response or severe by-effects.
15 cl, 25 dwg, 4 tbl, 15 ex
FIELD: chemistry, immonology.
SUBSTANCE: hybrid protein includes 936 protein sequence from Neisseria meningitides or protein with sequence identical to the mentioned protein sequence by 90% or more, and 741 protein sequence from Neisseria meningitides or protein with sequence identical to the mentioned protein sequence by 90% or more. The sequences can be linked by N- and/or C-end to histidine labels, with or without a linker. Linker is selected out of group of polyglycine linker, histidine labels and GSGGGG linker. A nucleic acid encoding this protein is displayed. Invention also claims composition for treatment and/or prevention of disease caused by Neisseria meningitides bacterium, based on hybrid protein and one or more proteins of the following group: 287, 741, ORF46.1, 961, NH2-A-[X-L-]n-B-COOH, where n=2, X1=287, and X2 is selected out group of: 953, 919, 961, 741. Invention claims application of composition in production of medicine for treatment of disease caused by Neisseria.
EFFECT: efficient treatment and prevention of disease.
13 cl, 5 dwg, 28 tbl, 1 ex
FIELD: biology, gene engineering.
SUBSTANCE: invention can be used for marking of biological objects. The molecule of nucleic acid which codes the fluorescing protein chosen from fluorescing proteins of representatives of kind Phialidium sp. are both suborder Anthomedusae and fluorescing mutants of the specified proteins allocated. By means of the allocated nucleic acid are obtained cloning and expressing vectors, fluorescing protein, the protein of merge capable to fluorescence, and also the expressing cartridge. The cell and the stable cellular line, containing such express ionic cartridge, produce fluorescing fiber. The fluorescing protein, nucleic acid coding it and the express ionic genetic designs containing this nucleic acid, use in a set for marking of a biological molecule. Fluorescent protein is also used in methods of marking of a biological molecule, a cell or a cellular organella.
EFFECT: invention application allows dilating an arsenal of agents for marking of biological objects.
13 cl, 12 dwg,12 ex
SUBSTANCE: invention refers to pharmaceutical set of cancerous disease treatment and prevention. The composition comprises cisdiammoniumdichlor-trans-dihydroxoplatinum (IV), in particular its salts, physically separated from the carrier selected from the group comprising tablet, capsule, coated pill, suppository, ointment, cream, solution for injection where the carrier is combined with cisdiammonium-dichlor-trans-dihydroxoplatinum (IV) immediately prior to use. The invention also refers to pharmaceutical composition obtained from the claimed set.
EFFECT: physical separation of active agent and carrier and their integration immediately before use enables to significantly increase antineoplastic activity of composition.
11 cl, 4 tbl
SUBSTANCE: invention relates to medicine, namely to oncology and can be applied for combined treatment of stomach cancer. Method includes operative treatment and drop introduction of 5-fluorouracil in portal vein. Twenty four hours before operation drainage of pectoral lymphatic duct in neck area is carried our, then before operation catheterisation of portal vein is carried out and pre-operation chemotherapy by 5-fluorouracil in doze 800-1000 mg is started. Before gastrectomy at the level of I-II of lumbar vertebras on the left of backbone pectoral lymphatic duct is separated and compressed for period of surgical interference. In post-operation period during twenty four hours drainage of pectoral lymphatic duct is continued.
EFFECT: invention allows increasing of combined treatment efficiency by carrying out operation in conditions of "lymphokinesis arrest" and excretion of metabolism products in draining of pectoral lymphatic duct.
1 ex, 3 dwg
SUBSTANCE: invented compounds have inhibitory activity towards protein kinase. In formula 1a m lies between 0 and 1, R1 is chosen from a group which includes hydrogen, methyl, isopropyl, imidazolylpropyl, piperazinylpropyl, pyridinyl, diethylaminopropyl, hydroxyethyl, pyrimidinyl, morpholinopropyl, phenyl, cyclopropyl, morpholinoethyl, benzyl and morpholino, where any of pyridinyl, imidazolyl, piperazinyl or pyrimidinyl in R1 are optionally substituted with 1-3 radicals, independently chosen from a group, which includes methyl, methylamine, dimethylaminomethyl, cycloproylamine, hydroxyethylamine, diethylaminopropylamine, pyrrolydinylmethyl, morpholino, morpholinomethyl, piperazinylmethyl and piperazinyl, where any of morpholino and piperazinyl in R1 are optionally further substituted with a radical, chosen from a group which includes methyl, hydroxyethyl and ethyl, R2, R3 and R5 each represents hydrogen, R4 represents methyl, L is chosen from a group which includes -NR5C(O)- and -C(O)NR5-, R10 represents trifluoromethyl, and R11 is chosen from a group which includes halogen, morpholinomethyl, piperazinyl, optionally substituted with a methyl, ethyl or hydroxyethyl group; piperazinylmethyl, optionally substituted with a methyl or ethyl group, imidazolyl, optionally substituted with methyl, pyrrolidinylmethoxy and piperidinyl, optionally substituted with a hydroxy group.
EFFECT: more effective treatment.
4 cl, 1 tbl, 3 ex
SUBSTANCE: invention refers to medicine, oncology, and can be used for treatment of patients with breast cancer. For that purpose course of autohemochemotherapy is carried out. Before carrying out autohemochemotherapy, action on the ulnar vein projection is performed every day with optical radiation of green light with wave 560 nm long and blue light with wave 450 nm long. At that, there used is frequency of 1.5 Hz - 1.7 Hz - 3.0 Hz - 3.4 Hz - 6.0 Hz -6.8 Hz - 7.8 Hz - 9 Hz with exposure time of 2 to 4 minutes for each frequency. At that, if level of leucocytes is less than 4.0×109/l, the exposure time of optical radiation is increased by 50% till the level of leucocytes is 4.0-8.0×109/l and percentage of lymphocytes in white cell count is 20% and more.
EFFECT: proposed method allows keeping total number of leucocytes of the patients within the norm, decreasing intense intoxication, which allows performing complete autohemochemotherapy, has intense stimulating and protective effect on hemopoietic and immunocompetent organs of seriously ill patients, contributes to development of adaptive responses of training and activation, which improve total and antitumoural resistance of the organism.
SUBSTANCE: invention refers to medicine and namely to oncourology and can be used for treatment of bladder tumours. The method is implemented in the following way: during the period prior to the operation there created is depot of cytostatic incubated with autoplasma in regional lymphatic nodes. For that purpose 50 ml of blood is taken from the patient, centrifugated at the speed of 1500 rpm during 40 min; 15 ml of the obtained autoplasma is added to cytostatic diluted in the proportion of 0.9% NaCl per 5 ml, and incubated during 1 hour at t=37°C. Then puncture of lymphatic vessel on the back of the foot is performed and cytostatic with autoplasma is injected; repeated injection in one and the same limb is carried out in 12 days.
EFFECT: use of invention provides the effect on tumour and its regional metastases owing to keeping the remedy in lymphatic bed for a long time and its being deposited in regional lymphatic nodes at low probability of complications.
SUBSTANCE: anti-cancer remedy obtaining method is based on coordination compound of bivalent platinum and in accordance with which the water solution containing humic substances is processed with the above platinum compound. As coordination platinum compounds, the compound of the following row is chosen: potassium tetrachloroplatinate, cis-dichlorodiamine platinum, hydrogen tetrachloroplatinate, potassium tetrabromoplatinate. Processing is performed under influence of wave radiation up to the value of content of high-molecular fraction of humic substances of not more than 5%. As wave radiation, there chosen is radiation in ultrasonic range of frequencies of 18 kHz to 66 kHz with power of 0.5 to 5 W/cm3. When radiation power is 5 W/cm3 and frequency is 22 kHz, processing is performed during 5-20 min. As wave radiation, there can be chosen radiation in microwave range of frequencies of 30 to 0.3 GHz and power of 0.5 to 50 W/cm3. When microwave radiation power is 0.5 W/cm3 and frequency is 2.45 GHz, the temperature of the processed product is maintained within 40-50°C during 30-90 min.
EFFECT: method provides obtaining more than one homogeneous product which is more stable during storage by decreasing relative content of high-molecular fraction.
6 cl, 5 ex, 5 tbl, 2 dwg
SUBSTANCE: there are disclosed polypeptide variants containing Fc-areas IgG, having amino acid modifications providing changed effector functions Fc in specified polypeptides. There is disclosed composition for antibody targeting on antigen, containing the specified polypeptide. There is described method for preparing the specified polypeptide. Also, there are disclosed the methods for treating V-cell tumour or a malignant disease characterised by V-cell expression of CD20, treating chronic lymphocytic leukosis, relieving the symptoms of the V-cell controlled autoimmune disease, treating a angiogenesis-associated disorder, treating HER2-expressing cancer, treating LFA-1-mediated involvement, treating IgE-mediated involvement wherein specified methods imply introduction to the patient of the therapeutically effective amount of said polypeptide.
EFFECT: higher clinical effectiveness.
63 cl, 6 ex, 13 dwg, 10 tbl
SUBSTANCE: invention refers to new compounds of formula I where R1 stands for phenyl; G stands for C1-C7-alkylene; Q stands for -NH-; and X stands for C1-C7- alkylene, or to its salts. In addition, the invention concerns a pharmaceutical composition, to application of compound of formula I as defined in claims 1-5 item, as well as the method for making the compound of formula I.
EFFECT: production of the new biologically active compounds inhibiting protein tyrosine kinase.
8 cl, 3 ex
SUBSTANCE: invention relates to group of the new compounds presented by structural formulas A-K ,
and their pharmaceutically acceptable isomers, salts, salvates and to polymorphous forms.
EFFECT: inhibitory action with respect to thrombin receptors.
20 cl, 1 tbl, 2 ex
SUBSTANCE: new 5-sulphanyl-4H-1,2,4-triazole derivatives of general formula I (meaning of radicals R1-R3 are indicated in the description of the invention), methods of their preparation by liquid-phase parallel synthesis and pharmaceutical composition are claimed.
EFFECT: claimed compounds display high affinity to some subtypes of somostatin receptors of the SST2 and SST5 subtypes and possibility of their usage for treatment of pathological states or diseases involving one or more of the given somostatin receptors
9 cl, 708 ex
SUBSTANCE: invention concerns area of medicine and concerns compositions and medicinal forms on the basis of Gastrinum, application and reception methods. The essence of the invention includes the bond of Gastrinum representing conjugates of fragments of amino-acid sequence of Gastrinum, possessing functional ability to contact Gastrinum/SSK receptor, with various carriers, including application amino-acid spacers, and application of bifunctional sewing agents, and also a method of treatment by the compositions including bond of Gastrinum, sick of diabetes.
EFFECT: advantage of the invention consists in action prolongation.
7 cl, 8 ex, 3 tbl, 2 dwg