Method for prophylaxis of oncological diseases, or infections mordibidized by bacteria or fungi and protozoa, or arteriosclerosis, or diabetes mellitus, or diseases mediated by delayed hyperresponsiveness reaction, or diseases mediated by somatic cell gene mutations (variants)

FIELD: medicine and veterinary.

SUBSTANCE: invention relates to method for prophylaxis of oncological diseases, or infections mordibidized by bacteria or fungi and protozoa, or arteriosclerosis, or diabetes mellitus, or diseases mediated by delayed hyperresponsiveness reaction, or diseases mediated by somatic cell gene mutations. In the first embodiment of invention blood extracellular DNA destroying agent, such as DNAase, is administered into blood. In the second embodiment agent, binding to blood extracellular DNA, such as anti-DNA antibody is administered into blood. According to the third embodiment enzyme altering of blood extracellular DNA chemical structure is administered into blood. According to the forth embodiment agent, stimulating synthesis and/or activity of endogenic deoxyribonuclease or agent stimulating synthesis of antibody binding to blood extracellular DNA are administered into blood.

EFFECT: effective method for treatment of abovementioned diseases without side effects when prolonged using of preparation affected on blood extracellular DNA.

7 cl, 11 tbl, 18 ex, 5 dwg

 

The invention relates to medicine and veterinary medicine and can be used for the prophylaxis of oncologic diseases or infections caused by bacteria, fungi and protozoa, or atherosclerosis, or diabetes, or diseases associated with hypersensitivity reaction of the delayed type, or diseases, developing as a result of mutations in the genes of somatic cells.

These diseases are collectively the major causes leading to disability and mortality. Currently it is generally accepted concept of the primacy of prevention interventions (prevention of disease progression or relapse) in relation to the treatment of already developed the disease or its recurrence. In accordance with this intensively used and constantly developed new methods of drug prevention development and recurrence of these diseases.

It is generally accepted that vaccination is an effective and reliable method of prevention and recurrence of infections, however, an effective vaccine currently have managed to create only for prevention of a very limited number of bacterial, parasitic and fungal infections, see G.Ada, A.Ramsey; Vaccines, Vaccination and the Immune Responce; Lippincott-Raven, NY, 1997. Against most clinically relevant of infecti is the only way to prevent at the moment is the way chemoprophylaxis. As examples chemoprophylaxis widespread infection caused by protozoa malaria drugs chloroquine and mefloquine, see the Merck Manual of Diagnosis and Therapy; 16thEdition.

In a known manner chemoprophylaxis is prophylaxis with antibiotics and sulfa drugs bacterial infections, see the Merck Manual of Diagnosis and Therapy; 16thEdition, as well as the prevention of fungal infections in patients with immunodeficiency drug Amphotericin, see the Prophylaxis and treatment of fungal infections associated with haematological malignancies. Sibel Ascioglu et.al., International Journal of Antimicrobial Agents, Volume 15, Issue 3, July 2000, Pages 159-168.

The primary method of drug treatment of atherosclerosis is therapy with statins inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A(HMGCoA) reductase inhibitors (Lovastatin, Provastatin and others)that suppress the synthesis of cholesterol in the body and leading to faster clearance of low-density lipoproteins from the plasma of the blood, which slows the development of atherosclerosis, see New Concepts and Paradigms in Cardiovascular Medicine: The Noninvasive Management of Coronary Artery Disease, K. Lance Gould, THE AMERICAN JOURNAL OF MEDICINE, Volume 104, June 22, 1998, 2s 17s.

Drug primary prevention of cancer with proven efficacy (except for the primary prevention of specific rare forms of cancer) have not been developed (Enzyme Induction and Dietary Chemicals as Aproaches to Cancer Chemoprevention: The Seventh DeWitt S. Goodman Lecture. Allan H. Conney, Cancer Research, vol.63, pp.7005-7031, November 1, 2003). To prevent the recurrence of cancer are widely used methods based on anticancer chemotherapy, immunotherapy and hormonal therapy, see Adjuvant treatment for colorectal cancer.,Van Laethem JL, Acta Gastroenterol Beig 2001 Jul-Sep 64: 263-7; Adjuvant therapy for breast cancer patients: treatment decision tree from a French cancer network, Bachelot T, et.al., Bull Cancer, 2002, Oct 89: pp.897-903 used in the so-called "adjuvant"mode.

Medicinal ways to prevent diabetes with proven efficacy as well not exist. Known attempts to prevent type I diabetes in high-risk groups with the help of insulin therapy, see Effects of insulin in relatives of patients with type 1 diabetes mellitus., New Engi J Med 2002 346 May: pp.1685-91, and type II diabetes in high-risk groups drugs biguanides, see Metfbrmin & lifestyle intervention prevent Type 2 diabetes: lifestyle intervention has the greater effect., Doggrell SA, Expert Opin Pharmacother, 2002, Jul 3: pp.1011-3.

The most common way to prevent reactions associated with delayed-type hypersensitivity, is a method based on therapeutic immunosuppressive drug cyclosporine, see Therapeutic Immunosupression, ed. A.W.Thomson, Ser. Immunology and Medicine vol.29, Kluwer Acad. Publishers, Dordrecht, 2001.

He ways there are drug disease prevention, developing as a result of mutations in the genes of somatic cells. (Youssoufian H, Pyeritz RE. Mechanisms and Consequences of omatic Mosaicism in Humans, Nature Reviews Genetics, 2002; 3:748-758).

Analyzing therapeutic properties of the known methods of prevention of the mentioned diseases, we can identify the following main disadvantages:

1. Methods exhibit toxicity or cause complications (Adjuvant chemotherapy of neoplastic diseases, therapeutic immunosuppression with cyclosporine, prevention drugs statins).

2. The methods are not sufficiently effective or ineffective in the long-term use (insulinomimetic, a biguanide, a statin, anticancer drugs, money chemoprophylaxis infections, cancer drugs, cyclosporine).

3. Methods are active within a narrow range of diseases, or even in relation to specific forms of the same disease.

Currently there are no methods that would allow to realize the prevention of the above diseases in the complex, and to prevent their recurrence. In this regard, there is no possibility to take any known technical solution for the prototype of the present invention.

All of the variants of the present invention laid the task of creating non-toxic and effective method for long-term preventive and anti-relapse when the change of diseases accompanied by changes in qualitative and/or quantitative composition of extracellular DNA in the blood, namely, cancer, infections caused by bacteria, fungi and protozoa, atherosclerosis, diabetes mellitus, allergic diseases associated with hypersensitivity reaction of the delayed type, diseases, developing as a result of mutations in the genes of somatic cells.

According to the first variant of the invention this problem is solved due to the fact that blood is administered an agent that destroys extracellular DNA blood; an agent that destroys extracellular DNA blood may be for life; as an agent that destroys extracellular DNA blood may be used for enzyme Tnkase;

according to the second variant of the invention this task is solved by the introduction into the blood agent that binds extracellular DNA blood; as an agent that binds extracellular DNA blood can be used anti-DNA antibodies;

according to the third variant of the invention this task is solved by the introduction into the blood of an enzyme that modifies the chemical structure of extracellular DNA blood;

according to the fourth variant of the invention this task is solved by the introduction into the blood of an agent that stimulates the synthesis and/or activity of endogenous deoxyribonuclease, or an agent stimulating the synthesis of antibodies, bind is their extracellular DNA blood.

The development and recurrence of the considered diseases accompanied by qualitative and quantitative changes of extracellular DNA blood, but known to the applicant authorities lack knowledge about the genetic repertoire of extracellular DNA in the blood of patients with this disease, the biological role of extracellular DNA in the blood in this disease and possible therapeutic effect its destruction to prevent the development and recurrence of these diseases.

As identified by the applicants, extracellular DNA blood of patients with this disease contains a unique qualitative and quantitative composition of the repertoire of genes and genetic regulatory elements, dramatically different from the repertoire of DNA, described in the human genome. In contrast to intracellular DNA, extracellular DNA blood of patients considered diseases mainly contains a unique human genes. The presence of bacterial extracellular DNA and extracellular DNA of fungi in the composition of the matrix of biofilms and in the blood plasma of an infected person.

It is established that extracellular DNA blood, including extracellular DNA of bacteria, fungi and parasites in these diseases contributes to their development and recurrence.

It is established that the destruction, as well as option is Katsia and binding of extracellular DNA blood in these diseases leads to preventive and preventive effects.

The above new features of the claimed invention, based on new thinking about the role of extracellular DNA in the blood plasma in the development of the considered diseases and their recurrence, allow to draw a conclusion on the compliance of the claimed method the criterion of "inventive step".

The claimed method is implemented as follows.

Materials and methods.

In the first embodiment as an agent that destroys DNA, was used bovine pancreatic Tnkase (Sigma), bovine pancreatic Tnkase (Samson-Med) and recombinant human Tnkase I (Dornase; Genetech). The solution Gnkazy for injection was prepared by dissolving the matrix solution Gnkazy in sterile phosphate buffer just before the introduction.

In the second embodiment, as an agent that binds DNA, used antibodies against DNA extracted from blood of patients with systemic lupus erythematosus according to the method Shuster A.M. (Shuster A.M. et.al., Science, v.256, 1992, pp.665-667). Such anti-DNA antibodies can not only communicate, but also to carry out the hydrolysis of DNA.

In the third embodiment, as an agent that modifies DNA, used bacterial Sss I Methylase (CpG Methylase), (New England Biolabs).

In the fourth embodiment, as agent for stimulating the synthesis and/or activity of endogenous biopolymers, linking, or destroying, or modifying imicheskij composition, and/or conformation, and/or polymerization of extracellular DNA blood without its destruction, used G-f actin antibody (Calbiochem). G-Actin is an inhibitor of the activity of endogenous Gnkazy I. Linking actin antibodies increases the activity of endogenous Gnkazy I.

DNA in blood plasma was allocated as follows: fresh (not more than 3-4 hours after collection) blood plasma with added anticoagulant (sodium citrate) was off on a cushion of Ficoll-Plaque Plus (Amersham-Pharmacia) at 1500 g for 20 minutes at room temperature. Plasma (1/2 of the total number) were carefully selected, not touching the rest of the cells on the pillow ficoll, and Unscrew at 10,000 g for 30 minutes, to get rid of debris cells and debris. The supernatant was collected without disturbing the precipitate, was added to 1% sarkosyl, 50 mm Tris-HCl, pH to 7.6, 20 mm EDTA, 400 mm NaCl and an equal volume mixture of phenol-chloroform 1:1. The obtained emulsion was incubated at 65°2 hours, then separated phenol-chloroform by centrifugation at 5000 g for 20 minutes at room temperature. The procedure of deproteinization phenol-chloroform was repeated the same way three times, after which the aqueous phase was treated with chloroform, and then diethyl ether. Separation from organic solvents produced by centrifugation at 5000 g for 15 minutes. To the obtained aqueous phase was added an equal volume of isopropanol and incubi the Wali over night at 0° C. After precipitation of nucleic acids was separated by centrifugation at 0°, 10000 g for 30 minutes. The precipitated nucleic acids were dissolved in buffer containing 10 mm Tris-HCl, pH 7,6, 5 mm EDTA, and put on a pillow of speed cesium chloride (1 M, 2.5 M, 5.7 M) in a centrifuge tube rotor SW60Ti. The amount of DNA was 2 ml, the volume of each step CsCl and 1 ml of the Ultracentrifugation was performed in the device L80-80 (Beckman) 3 hours at 250000 g. DNA was collected from the surface of the steps 5.7M on fractions. Faction deliberately 12 hours at 4°C. the Presence of DNA in the fractions was determined by agarose electrophoresis with visualization of DNA methyl-ethidium. The amount of DNA was determined spectrophotometrically (Beckman DU70) in a cell with a volume of 100 μl, shooting range from 220 to 320 nm.

The implementation of the method is illustrated by examples.

Example 1. Prevention of recurrence of malignant tumors.

Antibodies against DNA was isolated from blood of patients with systemic lupus erythematosus according to the method Shuster A.M. (Shuster A.M. et.al., Science, v.256,1992, pp.665-667). Such anti-DNA antibodies can not only communicate, but also to carry out the hydrolysis of DNA.

Group 1 - 10 mice inoculated with Ehrlich carcinoma (control). On the 10th day after tumor inoculation mice received a single intraperitoneal injection of cyclophosphamide at a dose of 200 mg/kg

Group 2 - 10 mice inoculated with Ehrlich carcinoma. On the 10th day of the settlement of the E. inoculation of tumor mice received a single intraperitoneal injection of cyclophosphamide at a dose of 200 mg/kg On day 15 after tumor inoculation, mice received intravenous injection of a fraction of human anti-DNA antibodies (IgG) at 200 μg per animal.

Group 3 - 10 mice inoculated with Ehrlich carcinoma. On the 10th day after tumor inoculation mice received a single intraperitoneal injection of cyclophosphamide at a dose of 200 mg/kg On day 15 after tumor inoculation, mice received intravenous injection of the fraction of non-specific human immunoglobulin (IgG) at 200 μg per animal.

Group 4 - 10 mice inoculated with Ehrlich carcinoma. On the 10th day after tumor inoculation mice received a single intraperitoneal injection of cyclophosphamide at a dose of 200 mg/kg On day 15 after tumor inoculation, mice received one intraperitoneal injection of cyclophosphamide at a dose of 200 mg/kg

Anti-relapse effect binding of extracellular DNA in the blood was determined by survival of the animals on the 30th day after inoculation of the tumor. In the first and the third and fourth groups, all animals died. In the second group survived 50% of the animals.

Thus, the use of an agent that binds extracellular DNA in blood plasma, inhibits the recurrence of a malignant tumor according to the claimed method. The efficiency of binding of extracellular DNA blood exceeds the effectiveness of anti-re-course chemotherapy cyclophos is an.

Example 2. Prevention of recurrence of malignant tumors.

Patient 57 years. Two years ago was diagnosed carcinoma of lung stage IV with bone metastases and brain. The patient received 2 courses of chemotherapy with Taxol, however, due to the absence of tumour response and acute toxicity refused to continue chemotherapy treatment. The patient was pneumectomy, and she was discharged under the supervision of the district oncologist. After 3 months, the patient developed symptoms of compression of the upper Vena cava. When re-hospitalization was diagnosed with a relapse of the tumor in the field of bronchus stump. From radiological and chemotherapeutic treatment the patient refused. With the consent of the patient she had been given intramuscular injections of bovine pancreatic Gnkazy at a dose of 400 mg per day (2 injections daily; 2000 ED Kunz 1 mg). The patient was discharged. Over the next 2 months gradually disappeared symptoms of compression of the upper Vena cava. The health of the patient has improved. At follow-up visits after 3, 6 and 12 months - the absence of clinical and radiographic signs of disease progression.

Reduced levels of extracellular DNA blood (WDNC) (ng/ml) patient prior relapse rate according to the claimed method, through 2, 3, 6 and 12 months after the start against the recurrent rate shown on the graph (figure 1).

Thus, the use of Gnkazy according to the claimed method provides anti-relapse effect in malignant tumors.

Example 3. Prevention of development of autoimmune diabetes.

Diabetes in NOD mice occurs due to the development of autoimmune destruction of b-cells of the pancreas. Mouse 1st group (10 females line NOD) at the age of 6 weeks were given intramuscular injections of dornase-alpha (Genentech) at a dose of 500 mcg/kg (4 times daily) for 4 weeks. Mouse 2-nd group (10 females line NOD) received intravenous injection of a fraction of human anti-DNA antibodies (IgG) at 200 μg per mouse at the age of 6, 7, 8, 9 and 10 weeks. 10 mice from the 3rd group received four intramuscular injections of phosphate buffer with a 6 to 10 week. Since 15 weeks every three days was measured by the glucose level in the blood of mice. If the glucose level exceeded 250 mg/DL at two subsequent measurements, it was observed the presence of diabetes. Estimated number of cases to 15, 18, 24 and 28 weeks mice.

The results are shown in table 1.

Table 1

The ratio of healthy and diseased mice (the First figure is the number of healthy animals in the group; the second number is the number of sick animals in the group):
15 weekweek 1824 n the dividing 28 week
Group 18/08/27/36/4
Group 28/07/36/45/5
Group 38/24/62/82/8

Thus, the destruction of the binding of extracellular DNA in blood plasma has a preventive effect in the development of autoimmune diabetes.

Example 4. The influence of various methods of destruction binding and modification of extracellular DNA on its pathogenic properties.

Mouse C57B1 has been vaccinated highly metastatic or nizkopotentsialnogo strain tumors LLC. On day 9 after inoculation, animals were euthanized and collected a total blood plasma of mice. The total fraction of extracellular DNA blood after extraction were stored at -20°in phosphate buffer.

Participated in experiment 7 groups of mice inoculated nizkotitanistym strain LLC.

Group 1 - 6 mice inoculated with nizkotitanistym strain LLC.

Group 2 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of extracellular DNA of mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh heparinized the bed of blood).

Group 3 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of the DNA of the mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh plasma). Before the introduction of the DNA sample was subjected to photochemical disinfection (Appendix 1 μm methylene blue followed by irradiation with red light for 10 minutes (˜60000 Lux).

Group 4 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of the DNA of the mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh plasma). Before the introduction of the DNA sample was mixed with 10 µg of anti-DNA antibodies.

Group 5 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of the DNA of the mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh heparinized blood). Before the introduction of the sample was added to 1 μg of the fragment And the plant toxin Ricin, and incubated 1 hour at 37°C. the Ricin is a member of the family RIP (proteins inactivating ribosomes) toxins, widely used on the I create immunotoxins. In addition to the ability to inactivate ribosomes these proteins have the ability to degenerate DNA. For the implementation of the toxic effect of the catalytic unit And toxins RIP type II must be delivered into the cell subunit Century In the absence of a subunit In a chain And not toxic, but polynucleotide-adenylylation activity chain And can be used for inactivation of DNA circulating in the plasma.

Group 6 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of the DNA of the mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh heparinisation blood. The DNA sample before the introduction was subjected to enzymatic methylation (I.Muiznieks et.al., FEBS Letters, 1994, v.344, pp.251-254).

Group 7 - 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of extracellular DNA mice inoculated nizkotitanistym strain LLC.

8 group 6 mice inoculated with nizkotitanistym strain LLC + intravenously twice (on the seventh and eighth day after inoculation) introduction the total fraction of the DNA of the mice inoculated with highly metastatic strain (0.05 µg DNA before the introduction was dissolved in 500 μl of fresh heparinized the bed of blood. The DNA sample before the introduction incubated in the presence of 200 ng/ml of dornase-alpha 30 minutes at 37°C.

Estimated number of metastatic nodes in the lungs on day 15 after inoculation.

The results of the experiment are shown in table 2.

Table 2
GroupNcp.
1to 12.0
222,5
314,1
415,5
515,1
612,3
713,3
813,5

Thus, extracellular DNA in the blood of mice with vysokokachestvenny strain enhances tumor metastasis less of a malignant tumor. Destruction, linking and modification of extracellular DNA blood hinder the process according to the claimed method.

Example 5. Prevention of experimental atherosclerosis.

In the experiment used 18 outbred rats aged 20 months. Under General anesthesia, the left external carotid artery was kanalirovanii embolectomies catheter and was applied mechanical injury of the endothelium of the common carotid artery. Over the next 6 days 6 control rats received 4 intramuscular inye the tion phosphate buffer, 6 rats in group 1 received intramuscular injections of dornase-alpha, and 6 rats in group 2 received 2 intravenous injections of anti-DNA antibodies in a dose of 600 µg (in the first and third day after the operation).

On the 12th day of the damaged carotid arteries were removed, fixed in formalin and profinsirovano. When serial analysis of histological sections were analyzed intensity subendothelial changes in the area of applied mechanical injury. In mice the 1st and 2nd groups of the total area formed "neointima" averages were respectively 60% and 40% less compared to control animals.

Thus, destruction and binding and modification of extracellular DNA in the blood inhibit the development of "early atherosclerotic changes according to the claimed method.

Example 6. Prevention of bacterial and fungal infection.

In the experiment used outbred mice weighing 23-25 g.

group 1 (30 mice) mice were injected in retroorbital sinus pathogenic bacteria Staphylococcus aureus VT-2003R dose of 1×1010bacterial cells in the mouse. Recombinant human dornase-alpha (Geneyech) was administered at a dose of 500 mcg/kg intraperitoneally after 2, 6, 10 and 14 hours after infection.

2 group (10 mice) mice were injected in retroorbital sinus pathogenic bacteria Staphylococcus aureus VT-2003R dose of 1×1010 bacterial cells in the mouse. Phosphate buffer was injected intraperitoneally after 2, 6, 10 and 14 hours after infection.

After the last injection of dornase of 24 mice of group 1 were divided into sub-groups 1A and 1B.

Subgroup 1A (8 mice) - 2 hours after the last injection of dornase the mice were injected intravenously extracellular DNA blood (or 0.1 μg per mouse)obtained from mice infected with Staphylococcus aureus VT-2003R dose of 1×1010bacterial cells in the mouse after 15 hours after administration of bacteria in retroorbital sinus.

Subgroup 1B (8 mice) - 2 hours after the last injection of dornase mice were injected intravenously extracellular DNA blood (or 0.1 μg per mouse) were obtained from mice infected with Candida Albicans in a dose LD50 in mice, 3 days after intravenous injection of mushrooms.

Estimated survival of animals after 32 hours after infection.

The results are shown in table 3.

Table 3

The survival rate of mice at various times after infection:
0 hour.2 hour.4 hour.6 hour.8 hour.12 hour.24 hour.28 an hour.32 an hour.
Group 1100%100%100%90% 90%80%50%40%30%
Group 200%100%70%60%50%40%30%20%20%
1a20%10%10%
1B50%50%40%

3 group (10 mice) mice were injected intravenously mushrooms, subcultivation of pathogenic clinical isolate of Candida Albicans in a dose of LD50. Recombinant human dornase-alpha (Geneyech) was administered in the dose of 1 mg/kg intraperitoneally twice a day for 2, 3 and day 4 after infection.

4 group (10 mice) mice were injected intravenously mushrooms subculturally pathogenic clinical isolate of Candida Albicans in a dose of LD50. Amphotericin b was administered at a dose of 20 mg/kg intraperitoneally twice a day for 2, 3 and day 4 after infection.

5 group (10 mice) mice were injected intravenously mushrooms subculturally derived from pathogenic konicheskogo isolate of Candida Albicans in a dose of LD50. As a negative control was administered phosphate buffer vnutribruchinnogo in day 2, 3 and day 4 after infection.

Estimated survival and weight of mice on day 7 after infection.

The results are shown in table 4.

Table 4

The survival rate of mice at various times after infection:
1 day3 day5 dayday 7
Group 3100%100%100%100%
Group 4100%100%100%100%
Group 5100%80%50%50%

In the group of 4 on a 7 day weight of animals was on average 20% less than in group 3. This suggests that when equal protective efficacy of dornase-alpha and amphotericin b, the latter showed higher toxicity.

Thus, extracellular DNA in the blood of infected animals has a reinforcing effect on the course of the infectious process, and its destruction in accordance with the claimed method has a preventive effect as in bacterial and fungal infection.

Example 7. Prevention of autoimmune diabetes and malignant tumors.

In the experiment used 20 female mice NOD and 20 females we is her line SN.

Diabetes in NOD mice occurs due to the development of autoimmune destruction of b-cells of the pancreas. Mouse 1st group (10 females line NOD) received intravenous injection of fractions of mouse G-f actin antibody (Calbiochem) at 200 μg per mouse at the age of 6, 7, 8, 9 and 10 weeks. 10 mice from the 3rd group received four intramuscular injections of phosphate buffer with a 6 to 10 week. Since 15 weeks every three days was measured by the glucose level in the blood of mice. If the glucose level exceeded 250 mg/DL at two subsequent measurements, it was observed the presence of diabetes. Estimated number of cases to 15, 18, 24 and 28 weeks of mice

The results are shown in table 5.

Table 5

The ratio of healthy and diseased mice (the First figure is the number of healthy animals in the group; the second number is the number of sick animals in the group):
15 weekweek 1824 week28 week
Group 19/18/26/46/4
Group 27/35/53/73/7

Tumors induced by intradermal injection of 3-methylcholanthrene (1 mg 3-methylcholanthrene dissolved in 50 µl of olive is about oil, on the mouse) at four weeks of age. Estimated number of mice developed tumors at 10 weeks after administration of the carcinogen.

Mouse 3-th group (10 females line AWN) received intravenous injection of fractions of mouse G-f actin antibody at 200 μg per mouse at the age of 5, 6, 7, 8, 9 and 10 weeks.

Mouse 4-th group (10 females line SN) received intravenous injection of phosphate buffer at age 5, 6, 7, 8, 9 and 10 weeks.

Determined the presence of tumors in animals at the age of 14 weeks.

The results are shown in table 6.

Table 6

The ratio of healthy and diseased mice (the First figure is the number of healthy animals in the group; the second number is the number of sick animals in the group):
week 14
Group 32/8
Group 44/6

Thus, the suppression of activity of their own endogenous inhibitors Gnkazy I has a preventive effect in the development of autoimmune diabetes and cancer.

Example 8. Preventing the spread of the mutant gene. A number of human diseases arise due to the development status of somatic mosaicism - the expansion of a mutant gene in a population of somatic cells (Youssounan H, Pyeritz RE. Mechanisms and Consequences of Somatic Mosaicism in Humans. Nature Reviews Genetcs 2002; 3: 748-758).

As a model for the development of somatic mosaicism has been studied the frequency of mutations of the HPRT gene in T-lymphocytes. The human HPRT gene (Chromosome Xq26) encodes a constitutively expressed, but not essential enzyme involved in the metabolism of purine bases. Cloning was performed according to the procedure described Bigbee W (W. Bigbee Et al., Mutation Res., 1998, v.397, pp.119-136). The clone was subjected to the peripheral blood lymphocytes of 8 patients receiving the three-week course immunostimulating drug therapy Neovir after surgical removal of the tumor. Of the 8 patients, 4 patients were receiving additional therapy with human recombinant Dnazol I(Genetech) (200 µg/kg intravenously 4 times a day for 3 weeks). The frequency of HPRT-deficient clones in the blood of patients receiving therapy Dnazol I, the average was 3 times lower than that in the blood of patients who received only immunostimulirutuyu therapy. The addition of extracellular DNA in the blood of patients who did not receive Tnkase, into the culture medium when the cloning of T-lymphocytes of patients receiving therapy Dnazol, increases the frequency of HPRT-deficient clones cloning of the latter.

Thus, the destruction of extracellular DNA of the patient's blood by enzyme Dnazol reduces the frequency of the mutant gene in the population and prevents the development of somatic is mosaicism according to the claimed method.

Example 9.

Profilaktika recurrence of the disease, Marchiafava-Micheli

Disease Marchiafava-Micheli is a very rare disease associated with the presence of somatic gene mutations in the gene locus PI (phosphatidyl glycan class A)Bolina O., 17 years old, suffers from this disease for 3 years. In the last 2 years the disease was able to compensate for the admission of steroid hormones. In the last year appeared again and began to intensively progressing symptoms: episodes of hemolysis (2-3 times per month), hemoglobinuria, hepatosplenomegaly, macrocytes, anisoles. Produced 4 blood transfusion. Four months ago, the patient developed an episode of acute renal failure (creatinine - 8.2 mg/DL) due to intense episodes of intravascular hemolysis in the background moved purulent angina. During the week the patient was conducted hemodialysis. After payment of the kidney with the consent of the patient it was prescribed weekly infusions of human donor DNA Asimov isolated from patients with systemic lupus erythematosus, in a dose of 500 mg (1, 2, 3 and 4 weeks). During this month, there was only one paroxysm of the disease (2 week) significantly lower intensity than the previous one. The crisis was easily cropped three-day course of prednisolone at a dose of 50 mg per day. In week 5 the pain is Oh were prescribed daily single intramuscular injection Gnkazy I dose 2000000 ED Kunz in the day. Over the next three months of observation, the patient was normalized formula blood disappeared yellowness and hepatosplenomegaly. The paroxysms of the disease was not repeated (see figure 2).

Example 10. Prevention of progression of odontogenic soft tissue infections

The study included patients with odontogenic abscesses and phlegmon of the soft tissues, entered the clinic of maxillofacial surgery. On admission, the patients were evaluated on a scale Ave. Patients with index Solovyova from 91 to 159 points, have a 50% chance of progression from infection after the start of treatment. Just for the study were selected 25 patients who had an index Solovyova from 91 to 159 points. In the control group treated with standard surgical benefits and antibiotic therapy was included 15 patients. In the experimental group, receiving more intramuscularly to Tnkase I in a daily dose of 1020000 ED. Kunz, was included 10 patients. On the third day after the start of treatment in the control group, 7 patients was detected local dissemination of the infection. Of the 10 patients of the experimental group, none of the local dissemination of the infection has not occurred.

Electropores extracellular DNA blood of patients with odontogenic infection of the soft tissues on day 3 from the start of treatment (figure 3).

Lane 1 - the Patient from the control group with local the th dissemination

The band 2 - the Patient from the experimental group

Band 3 - the Patient from the experimental group

Lane 4 - the Patient from the experimental group

Band 5 - Markers of molecular weight

Prevention of progression of malignant tumors.

The study included 9 patients with various forms of malignant tumors (breast cancer - 2; cancer of the stomach - 3; lung cancer - 4)received in the clinic of thoracic surgery after previous ineffective chemoradiation treatment. All patients at the beginning of treatment there were signs of fast growth of metastases (according to repeated CT scanning). All patients received a 30-day course of treatment with Dnazol I daily intravenously in the form of six 30-minute infusions per day at a daily dose of 2100000 units Kunz. During follow CT study, only two patients were identified progression of the disease. These same patients revealed an increase of extracellular DNA and blood was determined by high titers of antibodies to DNase I. all other patients has been a distinct decrease in the level of extracellular DNA in the blood (figure 4).

Table 7

Antibody titers to the desoksiribonukleaza in patients 3 and 7
Patient 3Patients who NT 7
1 week4 weekBreeding1 week4 weekBreeding
2,0892,127501,6212,16350
0,9891,1492501,2812,017250
0,3200,40112500,3831,7681250
0,1250,15462500,2071,1256250

Example 12.

Prevention of progression of diabetes and atherosclerosis

In continuing the study included 10 patients with a diagnosis of type 2 diabetes. All patients were previously transferred for treatment with recombinant human insulin for long periods due to the inability to provide normoglycemia with oral antidiabetic agents. The average age of patients was 54 years. The average duration of disease since diagnosis was 4.5 years. Five patients of the experimental group were prescribed intramuscular injections of bovine pancreatic Gnkazy at a dose of 200 mg per day (two injections per day) for 6 months. Examined the content of the glycosylated hemoglo is in the blood, index of atherogenic lipoproteins in the blood (as an indicator of activity of atherogenesis; it is known that the vascular complications of diabetes naturally arise due to the rapidly developing atherosclerosis of large arteries). Calculated daily insulin requirement (as an indicator of the activity of the underlying disease).

The results of the experiment are shown in Table 8

Table 8

The effect of treatment Dnazol on metabolic indices in the group of patients treated by the present method.
Before treatmentAfter 4 monthsAfter 8 months
Glycosylated hemoglobin (%)a 12.710,37,9
Index of atherogenic lipoproteins of blood7,316,14,56
The need for insulin U/kg of body weightof 1.340,870,63
The content of extracellular DNA blood; ng/ml17012590

Two patients from the experimental group before the study was made myocardial perfusion studies with thallium 201 in connection with suspected coronary atherosclerosis and "painless" form of coronary heart disease. For both of them patients revealed diffuse focal zone with abnormal accumulation of the radiopharmaceutical, evidence of the presence of coronary atherosclerosis. Patients were prescribed standard therapy (beta-blockers + nitrates). On completion of the study, patients were directed to re-perfusion scintigraphy of the myocardium. In the dynamics of a marked improvement in picture isotope accumulation in the myocardium, indicating the improvement of blood flow in the basin of the coronary arteries.

Example 13.

Prevention of recurrence of malignant tumors In September - October 2003 in the clinic of thoracic surgery, three patients (2 men aged 47 and 55 years of age and one female, aged 51 years) with a diagnosis of lung cancer in stage T4N2M+ were produced palliative surgery removal of the affected lung excision of solitary metastatic nodes in the opposite lung. All patients had previously received chemotherapy and radiation treatment. After surgery all patients with their consent had been given daily intramuscular injections deposited prolonged Gnkazy dose 5000000 Units Kunz in the day. To date (follow - up period was 10 months follow-up examination every 2 months) none of the observed patients not detected signs of recurrence of lung cancer. At the same time, these retrospective control suggests that the likelihood of relapse to Dunn the mu term and in this clinical situation is close to 100%.

Example 14. Prevention of bacteremia.

The study included 10 patients attending a dental clinic for acute dental pain. All patients were diagnosed with acute suppurative apical periodontitis and shows the removal of the diseased tooth. Patients were divided into 2 groups of 5 people. With the consent of the patients from the experimental group 30 minutes before the operation, they are intravenously once injected Tnkase I dose 2000000 Units Kunz. 2 hours after surgery in all patients was performed blood cultures. 5 patients experienced group seeding were sterile. In 3 patients of the control group of blood was inoculated Streptococcus.

Example 15. Prevention of hypersensitivity of the delayed type.

30 C57B1 mice were immunized with a suspension of Mycobacterium Smegmatis (100 μg of antigen in 50 µl aluminum alum) subcutaneously in the foot pad. After 4 weeks in the pad opposite the paws of mice were administered a resolving dose of antigen (50 μg).

Mice of the first group (10 mice), 30 minutes prior to the introduction of the resolving dose was administered anti-DNA antibodies in intravenous dose of 200 μg per mouse.

The mice of the second group (10 mice), 30 minutes prior to the introduction of the resolving dose was intravenously injected Enzymology containing bacterial Sss I Methylase (CpG Methylase, NewEngland Biolabs). In experiments Sss I Methylase included in the composition of small single-layer liposomes (SUV) in the ratio of 1u f is rment 1 µg lipids (Enzymology) at a dose of 750 μg per mouse.

Mice of the control group (10 mice), 30 minutes prior to the introduction of the resolving dose intravenously injected phosphate buffer.

Evaluated the intensity of edema of the legs 1, 2, 5 and 24 hours after administration of the resolving dose of antigen.

The results are shown in table 9.

Table 9

The intensity of the edema paw at different times after the introduction of the resolving dose of antigen.
30 minutes1 hour2 hours5 hours24 hours
Control2,13,63,74,14,2
Group 12,12,83,02.92,9
Group 22,13,03,13,43,0

Example 16. Prevention of recurrence of malignant tumors in animals

As an agent stimulating the synthesis of endogenous deoxyribonuclease, used stem cells from umbilical cord blood (CPCS), transferowania in vitro cDNA gene Deoxyribonuclease 1 person.

Group 1 - 7 mice inoculated with Ehrlich carcinoma - control.

Group 2 - 8 mice inoculated with Ehrlich carcinoma, received on the third day, p is after inoculation of the tumor intravenous injection of 25 million CPCS.

Group 3 - 8 mice inoculated with Ehrlich carcinoma, received on the third day after inoculation of the tumor intravenous injection of 25 million CPCS, transfected in vitro cDNA gene Deoxyribonuclease 1 person.

The effect was determined by the size of the tumor at day 7 after inoculation

Table 10

The size of the tumor 7 days after inoculation:
GroupTumor volume mm3
1117+/-12
2111+/-17
343+/-15

Example 17.

Prevention of the recurrence of a malignant tumor. In January 2004 at the clinic of thoracic surgery two patients (male aged 57 years and a woman aged 55 years) with a diagnosis of lung cancer in stage T4N2M+ were produced palliative surgery removal of the affected lung. All patients had previously received chemotherapy and radiation treatment. After surgery patients with their consent, was carried out the procedure for mobilization of stem cells in peripheral blood (GM-CSF 5 mcg/kg subcutaneously three times within 3 days) with subsequent isolation of stem cells by the method of cytapheresis apparatus Haemonetic. Selected cells were subjected to transfection with cDNA of the gene of human dornase-alpha (Desox the ribonuclease (I) method of cationic liposomes and injected intravenously in the amount of 50000000 cells. The procedure was repeated after 3, 6 and 9 months. To date (follow - up period was 14 months follow-up examination every 3 months) none of the observed patients not detected signs of disease progression. At the same time, these retrospective control suggests that the likelihood of recurrence to the given time and in a given clinical situation is close to 100%. Figure 5 provides information about the content of extracellular DNA in the blood of patients before treatment and during the course of treatment by the present method.

Example 18. Prevention of recurrence of malaria

Patient D., 49 years old, was admitted to the hospital on the 20th day from the onset of the disease with complaints of increasing temperature to 40-41°With stunning chills in the morning and afternoon hours, repeated after 48 hours, the duration 2.5-3 hours, changing the heat for 4 hours, without sweating, weakness, intense diffuse headache, poor appetite, sleep interrupted. 2 months ago came from India, where she spent the holidays. Microscopic examination of blood detected Plasmodium Vivax (3-5 in the field of view). The patient had been prescribed Delagil in exchange dose of 2.5 g and Primaquin 0.015 g per day in the course dose of 0.2, On the fourth day from the start of treatment with microscopy in the blood remained Plasmodium Vivax, continued bouts of fever and hepatosplenomegaly. the consent of the patient she had been implemented intradermal injection of DNA from calf thymus, mixed with incomplete adjuvant's adjuvant (50 mg/50 mg). Only 3 injections with an interval of 48 hours. Within 2 weeks had been symptomatic treatment. 2 weeks was a refresher course Delagila, after which it was observed sanitation blood parasite, the disappearance of bouts of fever and normalization of well-being.

The table lists information about the content of extracellular DNA in the blood of the patient and the titer of anti-DNA antibodies during treatment (Day 0 if admitted to hospital)

Table 11
Days from start of treatment048121418222630
in DNA blood ng/ml11712315513873461275
The titer of anti-DNA antibodies (at a dilution of 1:50)0,1130,1240,3540,6141,5342,2112,6552,6442,520

1. A method of preventing the development of cancer, or infections caused by bacteria, fungi and protozoa, or atherosclerosis, or diabetes dia is ETA, or diseases associated with hypersensitivity reaction of the delayed type, or diseases, developing due to mutations of genes of somatic cells, characterized by the fact that blood is administered an agent that destroys extracellular DNA blood.

2. The method according to claim 1, characterized in that the introduction of an agent that destroys extracellular DNA blood carry for life.

3. The method according to claim 1, characterized by the fact that as an agent that destroys extracellular DNA blood using the enzyme Tnkase.

4. A method of preventing the development of cancer, or infections caused by bacteria, fungi and protozoa, or atherosclerosis, or diabetes, or diseases associated with hypersensitivity reaction of the delayed type, or diseases, developing as a result of mutations in the genes of somatic cells, characterized by the fact that blood is administered an agent that binds extracellular DNA blood.

5. The method according to claim 4, characterized by the fact that as an agent that binds extracellular DNA, blood, use of anti-DNA antibodies.

6. A method of preventing the development of cancer, or infections caused by bacteria, fungi and protozoa, or atherosclerosis, or diabetes, or diseases associated with hypersensitivity reaction of the delayed type, or disease, is avivausa due to mutations of genes of somatic cells, characterized by the fact that the blood is injected enzyme that modifies the chemical structure of extracellular DNA blood.

7. A method of preventing the development of cancer, or infections caused by bacteria, fungi and protozoa, or atherosclerosis, or diabetes, or diseases associated with hypersensitivity reaction of the delayed type, or diseases, developing as a result of mutations in the genes of somatic cells, characterized by the fact that in the blood enter the agent stimulating the synthesis or activity of endogenous deoxyribonuclease, or an agent stimulating the synthesis of antibodies that bind the extracellular DNA blood.



 

Same patents:

FIELD: medicine and veterinary.

SUBSTANCE: invention relates to method for treatment of diseases, associated with alterations of blood extracellular DNA, such as generalized infections mordibidized by bacteria or fungi and protozoa, or arteriosclerosis, or diabetes mellitus, or diseases mediated by delayed hyperresponsiveness reaction, or diseases mediated by somatic cell gene mutations. Method includes administering of blood extracellular DNA destroying agent into blood. As such agent DNAase is used, in particular in doses providing alteration of electrophoretic profile of blood extracellular DNA, detectable by pulse gel electrophoresis. DNAase may be administrated in doses and regimes providing exceeded levels of blood plasma DNA-hydrolytic activity, namely 150 Kuntz/l of plasma, during 12 h/day in total.

EFFECT: effective method for treatment of abovementioned diseases without side effects.

4 cl, 14 tbl, 15 ex, 5 dwg

FIELD: medicine, cardiology.

SUBSTANCE: the complex of medicinal therapy includes nibentane to be injected at the dosage of 0.125 mg/kg intravenously. Moreover, one should pre-inject magnesium sulfate solution at the dosage of 2.5 g about 10-15 min before nibentane injection. The innovation provides quick restoration of sinus rhythm in case of no therapeutic complications observed.

EFFECT: higher efficiency of therapy.

3 ex

FIELD: organic chemistry, medicine, hematology.

SUBSTANCE: invention elates to new compounds that inhibit activated blood coagulating factor X (Fxa factor) eliciting the strong anti-coagulating effect. Invention proposes compound of the formula (1): Q1-Q2-C(=C)-N-(R1)-Q3-N(R2)-T1-Q4(1) wherein R1, R2, Q1, Q2, Q4 and T1 have corresponding values, and Q2 represents the group of the formula: wherein R9, R10 and Q5 have corresponding values also, or its salt, solvate or N-oxide. Invention provides the development of a novel compound possessing strong Fxa-inhibiting effect and showing the rapid, significant and stable anti-thrombosis effectin oral administration.

EFFECT: valuable medicinal properties of compounds.

13 cl, 1 tbl, 195 ex

FIELD: organic chemistry, medicine, cardiology, biochemistry.

SUBSTANCE: invention relates to benzoyl guanidines of the formula (I): wherein R1 means -CF3; R2 means -Y-para-(C6H4)-R11, -Y-meta-(C6H4)-R11 or -Y-ortho-(C6H4)-R11 wherein R11 means (C1-C9)-heteroaryl comprising two or more nitrogen atoms adjoining across nitrogen (N) atom; Y means oxygen atom; R3 means hydrogen atom; R4 means (C1-C4)-alkyl, and to their pharmaceutically acceptable salts. Indicated compounds elicit very high activity with respect to inhibition of Na+/H+ exchange and improved water solubility and therefore they can be used as anti-arrhythmic medicinal agents with cardioprotective component for prophylaxis of infarction and treatment of infarction and for treatment of stenocardia. Also, proposed compounds inhibit pathophysiological processes in arising disorders induced by ischemia, in particular, in treatment of cardiac arrhythmia induced by ischemia.

EFFECT: improved preparing method, improved treatment and prophylaxis, valuable medicinal properties of compounds.

17 cl, 2 ex

FIELD: medicine.

SUBSTANCE: method involves applying intracoronary phosphocreatine introduction into infarction-responsible artery when carrying out coronary angioplasty. Phosphocreatine solution is injected after reperfusing the infarction-responsible artery at constant volume rate of 0.1-4 ml/s with introduced phosphocreatine dose being equal to 0.5-4 g.

EFFECT: reduced myocardium necrosis zone; prevented cardiac insufficiency and cardiac rhythm disorders.

4 cl

FIELD: organic chemistry, chemical technology, medicine, biochemistry.

SUBSTANCE: invention relates to quinuclidine compounds of the formula (I) , its salts or their hydrates wherein R1 represents hydroxyl group; W represents: (1) -CH2-CH2-; 2) -CH=CH-, or 3) -C≡C-; HAr represents 5-10-membered aromatic heterocycle comprising 1-2 heteroatoms taken among nitrogen atom and sulfur atom that in addition to the group -X-Ar can be substituted with 1-3 groups taken among: (1) halogen atom; (2) (C1-C6)-alkyl, (C2-C6)-alkenyl or (C2-C6)-alkynyl group substituted optionally with: (a) hydroxy-group; (b) (C1-C6)-alkoxycarbonyl; (c) (C1-C6)-alkanoyl optionally substituted with (C1-C6)-alkoxy-group; (d) hydroxylated (C3-C8)-cycloalkyl; (e) (C1-C6)-alkoxy-group; (f) 5-6-membered aromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom, or (g) cyano-group; (3) (C1-C6)-alkoxy-group optionally substituted with: (a) hydroxy-group; (b) (C1-C6)-alkoxy-group optionally substituted with (C1-C6)-alkoxy-group; (c) halogen atom; (d) 4-6-membered nonaromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom; (e) 5-6-membered aromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom; (4) (C1-C6)-alkylthio-group optionally substituted with (C1-C6)-alkoxy-group or hydroxy-group; (5) 5-6-membered heterocyclyloxy-group comprising 1-2 oxygen atoms in heterocycle; (6) amino-group represented by the formula: -N(R3)R4 wherein R3 and R4 are similar or different and each represents hydrogen atom or group taken among: (a) (C1-C6)-alkyl group; (b) (C1-C6)-alkoxy-(C1-C6)-alkyl group; (c) carbonyl substituted with (C6-C14)-aryl; (d) (C6-C14)-arylsulfonyl or (e) 4-6-membered nonaromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom; (7) (C3-C8)-cycloalkyl or cycloalkenyl hydrocarbon group optionally substituted with: (a) oxo-group or (b) hydroxy-group; (8) (C6-C14)-aromatic hydrocarbon ring optionally substituted with: (a) (C1-C4)-alkylene dioxy-group or (b) hydroxy-group; (9) 5-6-membered aromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom optionally substituted with: (a) cyano-group or (b) (C1-C6)-alkoxy-group; (10) 4-6-membered nonaromatic heterocycle comprising 1-3 heteroatoms taken among nitrogen atom, sulfur atom and oxygen atom optionally substituted with one or some groups taken among: (a) hydroxy-group; (b) halogen atom; (c) cyano-group; (d) (C1-C6)-alkoxycarbonyl; (e) (C1-C6)-alkyl; (f) (C1-C6)-alkoxy-group that is optionally substituted with halogen atom or (C1-C6)-alkoxy-group; (g) (C1-C6)-alkanoyl; (h) (C1-C6)-alkoxy-(C1-C6)-alkyl; (i) oxo-group; (j) (C1-C4)-alkylenedioxy-group; (k) (C3-C8)-cycloalkylalkoxy-group or (C3-C8)-cycloalkenylalkoxy-group; (11) carbamoyl of the formula: -CO-N(R5)R6 wherein R5 and R6 can be similar or different and represent hydrogen atom, (C6-C14)-aryl wherein indicated aryl is optionally substituted with halogen atom, or (C3-C8)-cycloalkyl; or R5 and R6 form in common 3-6-membered ring; (12) carbonyl optionally substituted with (C1-C6)-alkoxy-group; X represents: (1) a simple bond; (2) (C1-C6)-alkylene chain; (3) (C1-C6)-alkenylene chain; (4) (C1-C6)-alkynylene chain; or (5) formula: -Q- wherein Q represents oxygen atom or sulfur atom; Ar represents: (1) (C6-C14)-aromatic hydrocarbon ring optionally substituted with one or some groups taken among: (a) halogen atom; (b) (C1-C4)-alkoxy-group or (c) (C1-C6)-alkylthio-group; or (2) 5-6-membered aromatic heterocycle comprising 1-2 heteroatoms taken among nitrogen atom and sulfur atom. Compounds of the formula (I) show inhibitory activity with respect to a squalene-synthesizing enzyme. Also, the invention relates to an inhibitor of squalene-synthesizing enzyme and the corresponding medicinal composition based on compound of the invention, a method for prophylaxis and treatment of disease wherein inhibition of squalene-synthesizing enzyme is effective. Also, invention proposes some methods for preparing compounds of the formula (I).

EFFECT: improved preparing method, valuable of medicinal and biochemical properties of com[pounds and composition.

25 cl, 10 tbl, 214 ex

FIELD: medicine.

SUBSTANCE: method involves administering a combination of an agent reducing cholesterol content in blood and reduced coenzyme Q10 of general formula .

EFFECT: enhanced effectiveness of treatment.

8 cl, 2 tbl

FIELD: medicine, cardiology.

SUBSTANCE: the present innovation deals with introducing nitrates, heparin, beta-blocking agents, calcium antagonists, aspirin. Additionally, one should intravenously inject dalargin once daily at the rate of about 5-7 mcg/kg/h at the dosage of 25-30 mcg/kg daily per 100 ml sodium chloride physiological solution for about 5-6 d against the onset of hospitalization period. The innovation provides favorable impact upon diastolic function of left ventricle by decreasing the risk of dangerous arrhythmias and coronary lethality.

EFFECT: higher efficiency of therapy.

2 ex

FIELD: medicine, cardiology.

SUBSTANCE: one should introduce the suspension of autologous mononuclear medullary cells without preliminary cultivation in to the mouth of coronary artery nourishing infarction area. Cell suspension should be introduced at the quantity of 100-150 mln. cells immediately after stenting coronary artery due to technique of passive passage. The procedure should be performed on the 14th - 21st d against the onset of the disease mentioned. The method provides efficient counterbalance of cardiomyocytes due to applying valuable stem cells at excluding complications induced by arterial occlusion.

EFFECT: higher efficiency of therapy.

1 ex, 1 tbl

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of triazaspiro[5,5]undecane of the formula (I):

wherein values of radicals R1-R5 are given in the invention claim, ort o their quaternary ammonium salts, N-oxides or nontoxic salts. Proposed compounds possess inhibitory and regulating activity with respect to chemokine/chemokine receptors and can be useful in prophylaxis and treatment of different inflammatory diseases, such as asthma, atopic dermatitis, nettle rash, allergic diseases, nephritis, hepatitis, arthritis or proliferative arthritis and other similar diseases. Also, invention relates to pharmaceutical compositions based on compounds of the formula (I).

EFFECT: improved control method, valuable medicinal properties of compounds.

9 cl, 5 sch, 36 tbl, 70 ex

FIELD: medicine, veterinary science.

SUBSTANCE: the present innovation deals with treating malignant tumors. For this purpose, its is necessary to provide a blood supply of an agent that destroys extra-cellular blood DNA. This agent should be introduced in dosages providing alteration of electrophoretic profile of extra-cellular blood DNA. Agent, also, should be introduced at the dosages and modes that provide the level of DNA-hydrolytic activity of blood plasma measured in blood plasma being above 150 Kunz units/l plasma during totally above 12 h daily. Therapy may last without intervals for 2 d, not less. As an agent destroying extra-cellular blood DNA one may apply DNAse, in peculiar case, bovine pancreatic DNAse or recombinant human DNAse. The innovation suggests, also, to apply and agent that binds extra-cellular blood DNA, for example, anti-DNA antibodies. The method provides low-toxic and efficient treatment of tumors, particularly at prolonged, even one's life-long therapy with preparations mentioned.

EFFECT: higher efficiency of therapy.

10 cl, 7 ex, 6 tbl

FIELD: medicine, oncology.

SUBSTANCE: the present innovation deals with treating gastric Helicobacter pylori-associated MALT-lymphomas. For this purpose, one should perorally introduce "Vitaflor" preparation as a ferment at daily dosage being not less than 40 g for 4 wk to an empty stomach or in intervals between meals, and in case of no complete regress of lymphoma at monotherapy with "Vitaflor" it is necessary to conduct additional courses of chemotherapy. The method provides cytostatic impact directly upon the tumor and suppresses proliferation of tumoral cells in case of complete absence of chemotherapy-accompanying side effects.

EFFECT: higher efficiency of therapy.

3 cl, 4 ex

FIELD: medicine.

SUBSTANCE: method involves administering local tumor cell radiomodification with following radiation therapy and radical surgical operation being applied. The local radiomodification is carried out by applying endovascular tumor tissue perfusion with 5% Metronidasole via superior rectal artery in superoampular rectal cancer cases and via superior rectal artery and one of internal iliac arteries in medial and inferoampular rectal cancer cases with said blood vessel occlusion done using non-lytic radiopaque emboli. Single remote radiation therapy is carried out at a dose of 10 Gy 1 h later after radiomodification being over. Surgical operation is done not later than in 24 h after the irradiation.

EFFECT: increased radiation therapy destruction effectiveness.

FIELD: organic chemistry, medicine, oncology, biochemistry, pharmacology.

SUBSTANCE: invention relates to the development of a method for inhibition in interaction of metalloproteinase 2 with integrin αvβ3 in host cells. Method involves contact of integrin with the amount of compound inhibiting this interaction and represented by the formula (I): wherein each of G1 and G2 represents independently -NH-C(O)-O-R1, -NH-C(O)-O-(CH2)v-(C6H4)-X3, -NH-C(O)-NH-(CH2)v-(C6H4)-X3, -O-C(O)-NH-(CH2)v-(C6H4)-X3, -O-C(O)-O-(CH2)v-(C6H4)-X3 or -NH-C(O)-CH2-(C6H4)-X3; each of Y1 and Y2 represents independently -OH, (C1-C4)-alkyl, (C1-C4)-hydroxyalkyl, (C1-C4)-alkoxy-group, phenyl, benzyl or -NH2; R1 represents (C1-C4)-alkyl; each of X1 and X2 represents independently halogen atom or (C1-C4)-alkoxy-group; X3 represents halogen atom, nitro-group, (C1-C4)-alkyl, (C1-C4)-alkoxy-group or (C1-C4)-perfluoroalkyl; Z represents -C≡C-, -C6H4-, cis-CH=CH-, trans-CH=CH-, cis-CH2-CH=CH-CH2-, trans-CH2-CH=CH-CH2-, 1,4-naphthyl, cis-1,3-cyclohexyl, trans-1,3-cyclohexyl, cis-1,4-cyclohexyl or trans-1,4-cyclohexyl; A represents hydrogen atom (H) or a covalent bond; each of m and n represents independently a whole number o or 1; t represents a whole number o or 1; each of p, r and v represents a whole number 1 or 2 and under condition that when A means hydrogen atom (H) then t is 0; when A means a covalent bond then t = 1, and when m = 0 then Y1 represents (C1-C4)-hydroxyalkyl, and when n = o then Y2 represents (C1-C4)-hydroxyalkyl. Also, invention describes a method for apoptosis induction involving administration of abovementioned substance in the therapeutically effective dose.

EFFECT: improved method for tumor inhibition, expanded assortment of antitumor agents.

37 cl, 9 dwg, 7 ex

FIELD: organic chemistry, amino acids, medicine, pharmacy.

SUBSTANCE: invention relates to using derivatives of cysteine for preparing a medicinal agent. The proposed agent is designated for treatment of diseases arising as a result of formation of heterotrimeric protein G, and to new derivatives of cysteine, and pharmaceutical composition based on thereof. Derivatives of cysteine, in particular, involve the following compounds: bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo-[2,2a]-pyrazine]-disulfide and bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-91-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo-[1,2a]-pyrazine-7-yl]-disulfide. Invention provides high effectiveness of treatment.

EFFECT: valuable medicinal properties of compounds.

6 cl, 7 dwg, 2 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: method involves carrying out radiation treatment combined with textile material application on the tumor. Tumor-transformed vaginal uterus neck portion volume is determined with ultrasonic examination techniques. Koletex napkin impregnated with therapeutic cytostatic preparation dose is used as the textile material. The napkin pattern is produced on the basis of ultrasonic examination data. The napkin is quilted with ligature along the perimeter and fixed at the level of vagina fornix by drawing tightly in purse-string mode. Napkin is changed every 24 h within 10-20 days long treatment course.

EFFECT: improved life quality.

FIELD: medicine, biology.

SUBSTANCE: invention relates to application of tissue factor agonist, namely FVII or FVIIa to induce or increase cell journey, as well as to application tissue factor antagonist modified with FVII to reduce or avoid cell journey in treatment of pathological conditions associated with specific control of cell journey or chemotaxis.

EFFECT: method for treatment of improved effectiveness.

14 cl, 14 ex, 10 dwg

FIELD: medicine.

SUBSTANCE: disclosed are combine product and kits useful in treatment for solid tumor by application of ZD6126 compound having formula in combination with platinum antitumor drug (e.g. cysplatine) and/or taxane. Treatment methods may include ionizing radiation. In was discovered that certain doses of abovementioned combined substances according to method of present invention have synergic action on solid tumors (as well as on angiogenesis neoplasm).

EFFECT: new method for solid tumor treatment.

12 cl, 5 tbl, 9 dwg

FIELD: medicine, oncology.

SUBSTANCE: one should carry out chemoradiation therapy at applying a cytostatic preparation followed by distance and intracavitary irradiation. Depending upon development of tumor lesion during the first 3 or 6 d it is necessary to conduct monochemotherapy only due to introducing proxiphen together with dimethyl sulfoxide at weight ratio of 4.5-5.0 : 0.5-1.5, correspondingly by applications in "Coletex" napkins. Moreover, a napkin should be pre-impregnated in 20%-dimethylsulfoxide solution and fixed with a tough vaginal tamponade by changing napkins every 24 h. Then since the 4th d or the 7th d simultaneously with application it is necessary to carry out contact irradiation and distance impact onto minor pelvis every 4-6 h at single focal dosage (SFD) being 2 Gy at 10 seances 5 times/weekly with high-activity sources of SFD 2 Gy. The innovation provides tumor regress under conditions of no therapeutic complications, thus, improving patients' quality of life.

EFFECT: higher therapy.

3 ex

FIELD: medicine, in particular angiogenesis prophylaxis and treatment.

SUBSTANCE: invention relates to 2-cyclooxygenase inhibitors selected from group containing 4-[5-(4-chlorophenyl)-3-phenyl-1H-pyrazole-1-yl] benzenesulfonamide; 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazole-1-yl] benzenesulfonamide; 4-[5-methyl-3-phenyloxazole-4-yl] benzenesulfonamide or pharmaceutically acceptable salts thereof and pharmaceutical composition containing the same in therapeutically effective amount. Said composition are useful in treatment and/or prophylaxis of angiogenesis disorders such as metastasis, eye angiogenesis, diabetic retinopathy, etc. in subjects are needed in such treatment and/or prophylaxis.

EFFECT: new pharmaceuticals for angiogenesis treatment and/or prophylaxis.

5 cl

FIELD: medicine and veterinary.

SUBSTANCE: invention relates to method for treatment of diseases, associated with alterations of blood extracellular DNA, such as generalized infections mordibidized by bacteria or fungi and protozoa, or arteriosclerosis, or diabetes mellitus, or diseases mediated by delayed hyperresponsiveness reaction, or diseases mediated by somatic cell gene mutations. Method includes administering of blood extracellular DNA destroying agent into blood. As such agent DNAase is used, in particular in doses providing alteration of electrophoretic profile of blood extracellular DNA, detectable by pulse gel electrophoresis. DNAase may be administrated in doses and regimes providing exceeded levels of blood plasma DNA-hydrolytic activity, namely 150 Kuntz/l of plasma, during 12 h/day in total.

EFFECT: effective method for treatment of abovementioned diseases without side effects.

4 cl, 14 tbl, 15 ex, 5 dwg

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