Angiogenesis inhibitor, anti-angiogenic pharmaceutical composition based on thereof and method for treatment of malignant neoplasm
FIELD: chemistry of natural compounds, biotechnology, pharmacology, medicine, oncology, pharmacy.
SUBSTANCE: invention relates to a medicinal formulation of angiogenesis inhibitor comprising active compound as liposome form wherein angiostatin or endostatin are used as an active component, and their truncated peptide fragments also, and an anti-angiogenic pharmaceutical composition used for treatment. Also, invention proposes a method for inhibition of tumor growth and metastasizing of malignant neoplasms associated with disorders in angiogenesis. Method involves administration of nontoxic therapeutically effective dose of pharmaceutical composition separately or in combination with adjuvant therapy with other antitumor preparations. Invention provides reducing rate of elimination of active compound (angiostatin, endostatin or their truncated fragments) from body and enhancing effectiveness of its therapeutic effect. Invention can be used for inhibition of tumor growth and metastasizing of malignant neoplasms.
EFFECT: enhanced and valuable medicinal properties of inhibitor.
7 cl, 3 tbl, 5 ex
The present invention relates to the field of chemistry of natural compounds, biotechnology, pharmacology and medicine and can be used for therapy of malignant tumors.
The invention relates to a liposomal form antiangiogenic drug for the treatment of malignant tumors, namely for the treatment of primary tumors and metastases in humans, for the treatment of malignant tumors and other pathologies associated with impaired angiogenesis. The advantage of the invention is to enhance antiangiogenic and antitumor activity of the active component of the drug.
Angiogenesis is the process of formation of new blood vessels from preexisting playing a key role in a wide spectrum of physiological and pathological processes. The progression of tumors and for cancer are closely linked to angiogenesis [L.A. Liotta et al., Cell 64: 327-336, 1991; I.J. Fidler and Ellis, L.M., Cell 79: 185-188, 1994; Folkman J., N.Engl. J.Med. 333: 1757-1763, 1995]. Developing the tumor through the secretion of angiogenic factors attracts new capillaries, emerging from nearby vessels. Further progression of the tumor due to the close integration of tumor cells and endothelium of blood vessels. Bring blood oxygen and nutrients cause rapid tumor progression and initiate the process of metastasis. When using antiangiogenic agents that cause the destruction of tumor blood network, in model experiments on animals with transplanted tumors by a number of authors observed a significant therapeutic effect of these drugs in terms of the suppression of tumor growth and the increase in the average life span of experimental animals [M.S. O'reilly et al., Cell 79: 315-328, 1994; M.S. O'reilly et al., Nat. Med. 2: 689-692, 1996; Holmgren L. et al., Nat. Med. 1: 149-153, 1995; Mauceri H.J. et al., Nature 394: 287-291, 1998]. Thus, the destruction of tumor blood network using antiangiogenic agents is a promising approach to therapy of malignant tumors.
One of the most powerful inhibitors of angiogenesis is angiostatin is a proteolytic fragment of plasminogen mol. weight ˜ 40 kDa [M.S. O'reilly et al., Cell 79: 315-328, 1994]. Acting on the endothelial cells of blood vessels, angiostatin inhibits the growth of both primary tumors and regional metastases.
Currently the tools of genetic engineering obtained biologically active recombinant angiostatin, expressed in different strains-producers (Escherichia coli, Pichia pastoris, and others). Recombinant angiostatin induces apoptotic death of endothelial cells and effectively suppresses tumor growth of lung carcinoma Lewis mice at daily doses of 1-6 mg/kg [WuZ. et al., Biochem. Biophys. Res. Commun. 236: 651-654, 1997]. One of the important features of angiostatin is its ability to suppress tumor growth without toxic effects [M.S. O'reilly et al., Nat. Med. 2: 689-692, 1996; Sim, B.K. et al., Cancer Res. 57: 1329-1334, 1997; Lannutti B.J. et al., Cancer Res. 57: 5277-5280, 1997]. Even with the drug in high doses (˜100 mg/kg of body weight) does not show any toxicity or the development of drug resistance of tumors to the drug [Sim, B.K. et al., Cancer Res. 57: 1329-1334, 1997; Mauceri H.J. et al., Nature 394: 287-291, 1998; Drixler T.A. et al., Cancer Res. 60: 1761-1765, 2000].
Despite the obvious benefits and prospects for therapeutic application of angiostatin, a significant disadvantage of the drug is its rapid elimination from the body [Drixler T.A. et al., Cancer Res. 60: 1761-1765, 2000]. In addition, to achieve the desired anticancer effect requires prolonged use of the drug. Thus, the drug is administered to the patient at least twice a day in doses of from 20 to 100 mg/kg of body weight. It is obvious that therapeutic application of angiostatin will not only cause inconvenience to the patients due to the need for regular injections, but will require a huge amount of preparation. In order to avoid this, it is necessary to reduce the rate of excretion of angiostatin and increase the efficiency of its therapeutic action.
Other promising antiangiogenic what Reparata, influencing the survival of endothelial cells, is endostatin - 20 kDa fragment of collagen XVIII, is a component of basement membranes and extracellular matrix [Y. Muragaki et al., Proc. Natl. Acad. Sci. USA, 92: 8763-8767, 1995]. Endostatin specifically inhibits proliferation and migration of capillary endothelial cells and can induce apoptosis in proliferating endothelial cells. Direct effect on tumor cells of various lines in vitro, endostatin does not [M.S. O'reilly et al., Cell: 88, 277-285, 1997]. In vivo, endostatin has a strong inhibitory effect against the growth of various primary tumors and their metastases. Prolonged therapy with high doses of endostatin causes almost complete blockade of tumor angiogenesis and leads to regression of the tumor to microscopic dimensions [Boehm .et al., Nature: 390, 404-407, 1997]. Further micropohone fall into "dormant" state, in which the index of proliferation of tumor cells is balanced by their index of apoptosis. Even after prolonged therapy is not observed the emergence of resistance to the drug or any toxic reactions [Sim, B.K. et al., Cancer Metastasis Rev.: 19, 181-190, 2000]. Endostatin can inhibit induced angiogenic growth factors cascade of intracellular signaling, as well as to block the activation and catalytic activity of the matrix metalloproteinases.
Closest to the technical nature are angiogenesis inhibitors angiostatin, endostatin and shorter fragments for the treatment of malignant tumors /Pat. Of the Russian Federation No. 2240328/. The disadvantages of these inhibitors described above. In addition, the authors of the present invention offer the use of the above inhibitors of angiogenesis in the form homodimeric proteins, in which the molecule inhibitor linked by a polypeptide of the bridge with the Fc region of immunoglobulin gamma, which may further reduce the biological activity of the target an inhibitor of angiogenesis. The use of such fused proteins also does not solve the problem of high rate of excretion of drugs from the body of the patient.
The purpose of the invention is the decrease in the rate of excretion of the active compound (angiostatin and endostatin) from the body and increase the efficiency of its therapeutic action.
The goal is solved by the creation of an inhibitor of angiogenesis, comprising the active compound in the form of liposomes. The active compounds are used angiostatin or endostatin, as well as their shorter peptide fragments. Also available antiangiogenic pharmaceutical composition for the treatment of malignant tumors and other pathologies associated with impaired angiogenesis containing the active compound (Academy of Sciences of histatin, endostatin or shortened fragments) and a pharmaceutical carrier, where the active compound is contained in a liposomal form in an effective amount, and as a pharmaceutical carrier are acceptable solutions for injection.
In addition, a method for treating malignant tumors and other pathologies associated with impaired angiogenesis, which consists in the introduction of a non-toxic therapeutically effective amount of the pharmaceutical composition, either alone or in combination with adjuvant therapy with other anticancer drugs, immunomodulators or antiangiogenic agents. As an anticancer drug for adjuvant therapy using a drug selected from the group of: doxorubicin, karminomitsin, mutamycin, Novantrone, rubomycin, bleocin, cisplatin, Taxol, etoposide, vinblastine, vincristine, Gemzar, fluorouracil, methotrexate, xeloda, Zoladex, dacarbazine, cisplatin, carboplatin, tamoxifen. As an immunomodulator for adjuvant therapy using a drug selected from the group of interferon-αinterferon-γ, interleukin-2, interleukin-4, interleukin-6, interleukin-12. As an antiangiogenic agent for adjuvant therapy using a drug selected from the group of thalidomide, vitaxin, pentosan, suramin, fumagillin, squal the min, combretastatin, prinomastat, marimastat, neovastat.
Angiostatin or endostatin in liposomes can be presented in various forms, differing in length of the peptide chain as whole proteins, and their shorter peptide fragments that differ from the sequence of the whole protein at one or more terminal amino acid. Angiostatin or endostatin for liposomal compositions can be obtained both from natural sources and with the help of genetic engineering technology, and shorter fragments of these proteins are also using genetic engineering methods or by the method of solid-phase peptide synthesis.
Created liposomal form of the inhibitor of angiogenesis is characterized by simplicity and adaptability of receipt shows high biological activity and can effectively suppress the growth of tumor blood vessels and tumors.
The basis of the invention is an inhibitor of angiogenesis, which is in the form of liposomes, which in experiments both in vitro and in vivo is higher antitumor activity than free active component. Inhibitor in the form of liposomes can be easily obtained using standard biotechnological and chemical methods, offers stability and adaptability of receipt. Liposomal form antiangiogenic inhibitor can be successfully used is implemented to treat a number of pathologies, associated with impaired angiogenesis. Thus, the invention fully meets the criterion of "inventive step".
The invention is illustrated by the following examples.
Example 1. Obtaining liposomes loaded with angiostatin or endostatin, or shorter fragments.
1 ml of liposomal dispersion in a round bottom flask is placed 20.5 mg of egg phosphatidylcholine (AFH) and 4.5 mg of cholesterol (AFH/cholesterol 7/3 mole.) in chloroform solutions. A mixture of solutions of lipids evaporated on a rotary evaporator to constant weight at 38°C. the Residues of the solvent is removed in vacuum oil pump within 2 hours of the Obtained lipid film was dissolved in 3 ml of ether and added dropwise into the flask 1 ml of protein solution (angiostatin, endostatin or their fragments) (6 mg/ml) in phosphate-buffered saline (pH 7.4). The flask was rinsed with argon and speak the dispersion in an ultrasonic bath for 3 min at 10°C. Carefully evaporated the ether on a rotary evaporator at room temperature (200 rpm). The obtained lipid dispersion push 19 times through nuclear polycarbonate filter with a pore size of 100 nm. Get 0.9 ml of a dispersion of liposomes of 100 nm (25 mg/ml lipid, 6 mg/ml protein). The effectiveness of the inclusion of protein is 14-26%. Liposomal inhibitor of angiogenesis lyophilizer and stored at 4°C.
Example 2. Getting formcomponents and on the basis of liposomal inhibitor of angiogenesis.
Pharmaceutical composition for injection is obtained by dissolving an effective amount of lyophilized inhibitor of angiogenesis, obtained as described in example 1 in physiological saline or phosphate-saline solution, prepared with water for injection. the medium pH is about 7.4. The concentration of the active substance (a protein) in a solution of 5-10%.
Example 3. Determination of antitumor activity of liposomal inhibitors of angiogenesis, including angiostatin, endostatin or shorter fragments in the individual therapy of mice with transplanted solid tumors melanoma B16.
Melanoma cells lines B16 maintained by weekly intraperitoneal inoculation of mice C57B1/6. For the induction of solid tumors the mice were injected subcutaneously cell suspension (2·105cells in 0.1 ml of physiological solution). Drugs were injected into experimental animals intravenously. A single dose of liposomal forms of the inhibitor of angiogenesis contained: 90 mg/kg angiostatin, endostatin or their fragments. The drugs were injected intravenously once a week, just three injections, starting from the 10th day after the inoculation of the tumor. Each experimental group consisted of six animals, control of ten. The size of the solid tumors were measured once in 2-3 days. Tumor volume was calculated by the formula where a - short; b - the longest diameter of the tumor. The relative size of the tumors (ORO) was determined by the formula:where Ropthe average size of tumors in the experimental group, Ptothe average size of tumors in control animals. The increase in life span of treated animals compared to the control was determined by the formula:where T is the average life expectancy (ALE) treated animals days From - ALE control animals, days.
The results of the experiment are given in table 1. Compared to the mild effect of the application liposonix forms of angiostatin, endostatin and its truncated fragments therapy liposomal forms has led to a significant inhibition of tumor growth both during treatment and after its abolition and the increase in the average life span of the treated animals.
The effectiveness of antitumor activity of liposomal inhibitor of angiogenesis in relation to melanoma In 16 mice in comparison with liposome forms.
|The group of animals||ORO, %||USPS, %|
|24 day||35 day|
|Liposome forms of medicines||angiostatin||64,8||82,4||20,8|
|angiostatin shortened||32,6||43,8||to 49.9|
Example 4. Antimetastatic activity of liposomal inhibitors of angiogenesis.
To evaluate the antimetastatic activity of liposomal antiangiogenic inhibitors, we conducted an experiment on mice with subcutaneously transplanted tumor Lewis lung carcinoma (3LL) in the background of the removal of the primary tumor site.
The primary tumor was removed on day 8 after inoculation. As the anesthesia used geksenal. Tumor site was deleted along with adjacent areas of the skin, for p is eupresidency the possibility of recurrence. The defect of the skin was closed by the overlapping seam, smeared with a 5% alcohol solution of iodine. The first introduction of the studied drugs was performed for 12 hours prior to surgery. The drugs were injected intravenously in doses of 90 mg/kg 1 time in 4 days, just 5 injections. Each experimental group consisted of 8 animals.
Antimetastatic effect was evaluated at the beginning of the loss of control animals. The intensity of metastasis was evaluated by comparing the average mass of lung metastases in the control and experimental groups of animals. At the same time used conditional intensity indicators of metastasis - the number and size of metastatic nodes in the lung tissue, the frequency of metastasis of the tumor. Statistical processing of the obtained results was performed by the t-test.
The growth inhibition of metastases was calculated by the formula:where Lto- average weight of the lung with metastases in the control group, Labout- average weight of the lung with metastases in the group treated mice.
The results of the experiment are presented in table 2. Compared with the control animals, in which the lung metastases detected in 100% of cases in the groups treated animals was observed in the absence of metastases and reduced the intensity of metastasis. Thus the m the application of the proposed drug for the prevention of metastasis is very promising.
Influence of liposomal inhibitors of angiogenesis on the intensity of metastasis subcutaneously inoculated tumor Lewis lung carcinoma (3LL) in the background of the removal of the primary tumor site.
|The group of animals||TRM, %||The frequency of metastasis||The intensity of metastasis (the average number of metastases/mouse)|
|liposomal form of angiostatin||42,7||5/8||[2,7±1,3]|
|liposomal form of angiostatin (abridged)||38,2||4/8||[2,1±1,9]|
|liposomal form of endostatin||40,6||5/8||[2,2±0,8]|
|liposomal form of endostatin (abridged)||34,6||5/8||[1,9±0,6]|
1. The data are given at the time of death.
2. In the column "Frequency of metastasis" is number of animals with metastases/number of animals in the group.
3. In gra the e Intensity metastasis" (in square brackets) - the average number of metastatic colonies in the lungs of the animal.
Example 5. The effectiveness of antiangiogenic pharmaceutical compositions, including angiostatin, endostatin or shorter fragments, in relation to melanoma mice in their application in the mode of combination therapy with anticancer drugs, immunomodulators and other antiangiogenic agents.
Antiangiogenic pharmaceutical composition was injected as described in example 3, on the background of adjuvant therapy with other drugs. Thus, animals of group 1 were additionally introduced antitumor antibiotic doxorubicin (2 mg/kg) in the same pattern as liposomal preparations, group 2 - interferon-α daily for 3 weeks, group 3 - angiogenic drug thalidomide. The results of the experiment are given in table 3. As can be seen from the above data, the application of the proposed angiogenic pharmaceutical compositions in the modes of combination therapy with other drugs significantly increases the effectiveness of their anti-tumor activity.
The effectiveness of the antitumor activity of antiangiogenic pharmaceutical compositions against melanoma line B16 mice when combined with other therapy and drugs.
|The group of animals||ORO, % (35 day)||USPS, %|
|Individual therapy||Liposomal form of angiostatin||48,9||41,8|
|Liposomal form of angiostatin (abridged)||43,8||to 49.9|
|Liposomal form of endostatin||42,8||47,3|
|Liposomal form of endostatin (abridged)||37,1||52,1|
|Doxorubicin||Liposomal form of angiostatin||41,6||49,8|
|Liposomal form of angiostatin (abridged)||37,7||of 56.4|
|Liposomal form of endostatin||38,4||55,2|
|Liposomal form of endostatin (abridged)||32,6||of 58.9|
|Interferon-α||Liposomal form of angiostatin||46,1||46,2|
|Liposomal form of angiostatin (abridged)||39,8||54,9|
|Liposomal form of endostatin||39,7||51,2|
|Liposomal form of endostatin (koroch the config)||35,3||56,0|
|Thalidomide||Liposomal form of angiostatin||40,2||52,9|
Similar results were obtained in experiments to study the effectiveness of the claimed angiogenic pharmaceutical compositions in the mode of combination therapy with other anticancer drugs - karminomitsin, mutamycin, Novantrone, rubomycin, bleocin, cisplatin, Taxol, etoposide, vinblastine, vincristine, Gemzar, fluorouracil, methotrexate, xeloda, Zoladex, dacarbazine, cisplatin, carboplatin, tamoxifen; immunomodulators interferon-γ, interleukin-2, interleukin-4, interleukin-6, interleukin-12; antiangiogenic agents - vitaxin, pentosan, suramin, fumagillin, squalamine, combretastatin, prinomastat, marimastat, neovastat.
Proposed by the applicant approach is devoid of the aforementioned drawbacks and combines the simplicity and adaptability of receiving liposomal forms of antiangiogenic drugs with high efficiency antitumor activity due to an increase in the residence time of the drug in the blood and its selective accumulation in the area of tumor blood vessels.
1. Dosage form of the inhibitor of angiogenesis, comprising as active substance angiostatin or endostatin in Linden is som characterized in that the active substance it contains a full-size form or a shortened peptide fragment of the indicated inhibitors, when this substance is enclosed in liposomal membrane based on phosphatidylcholine and cholesterol levels when a molar ratio of 7:3, the obtained liposome containing an active substance not less than 19 wt.%.
2. Antiangiogenic pharmaceutical composition for the treatment of malignant tumors, containing the active ingredient and the pharmaceutical carrier, characterized in that the active substance it contains a dosage form according to claim 1 and saline or phosphate-saline solution, and the concentration of the active substance in the solution is 5-10% for protein.
3. Method of inhibiting tumor growth and metastasis of malignant tumors associated with impaired angiogenesis, characterized in that the patient is given a non-toxic therapeutically effective amount of the pharmaceutical composition according to claim 2.
4. The method of inhibition according to claim 3, characterized in that it further conduct of combined adjuvant therapy with other anticancer drugs, immunomodulators or antiangiogenic agents.
5. The method according to claim 4, characterized in that as an anticancer drug for adjuvant therapy using repeat, selected from the group doxorubicin, karminomitsin, mutamycin, Novantrone, rubomycin, bleocin, cisplatin, Taxol, etoposide, vinblastine, vincristine, Gemzar, fluorouracil, methotrexate, xeloda, Zoladex, dacarbazine, cisplatin, carboplatin, tamoxifen.
6. The method according to claim 4, characterized in that as an immunomodulator for adjuvant therapy using a drug selected from the group of interferon-αinterferon-γ, interleukin-2, interleukin-4, interleukin-6, interleukin-12.
7. The method according to claim 4, characterized in that as an antiangiogenic agent for adjuvant therapy using a drug selected from the group thalidomide, vitaxin, pentosan, suramin, fumagillin, squalamine, combretastatin, prinomastat, marimastat, neovastat.
FIELD: biopharmacology, preparative biochemistry and medicine.
SUBSTANCE: the method deals with homogenizing mammalian tissues and centrifuging. It is necessary to add ammonium sulfate to supernatant that contains the complex of heat shock proteins (HSP) up to 35-65% saturation, the residue developed should be dissolved in phosphate buffer at pH 7.2 followed by chromatography upon a column with sephadex G-50, then obtained protein fraction should be exposed to the action of chromatography upon heparin-sepharose. Fractions that contain TSP 70, 90, 96 the content of which was detected with the help of corresponding antibodies should be united, concentrated and salted out due to ultrafiltration. The innovation provides more simplified and shortened terms for implementing the method without applying expensive equipment. The complexes obtained should be applied in immunotherapy of oncological diseases.
EFFECT: higher efficiency.
2 dwg, 1 tbl
FIELD: medicine, chemico-pharmaceutical industry.
SUBSTANCE: the present innovation deals with separate introduction of aqueous tincture of shelf fungus, propolis and honey for oncological patients, moreover, shelf fungus should be applied as aqueous tincture at a certain ratio, propolis should be applied as propolis butter, moreover, additionally, one should introduce ursine fat, the tincture of aspen bark, fir oil, fruit juice of viburnum or mountain ash, moreover, patients should take components separately, successively between meals in certain order and certain quantity, moreover, the constituents as tinctures and fruit juices should be prepared just before usage and propolis oil - beforehand. The above-mentioned technique favors efficient treatment of oncological patients.
EFFECT: higher efficiency of therapy.
2 cl, 2 dwg, 4 ex, 1 tbl
FIELD: medicine, oncology.
SUBSTANCE: invention relates to therapy of malignant neoplasms. Method involves administration of ascorbic acid or ascorbic acid glycoside in the dose 50 mg/kg or water-soluble vitamin E in the dose 100 mg/kg. These preparations are administrated simultaneously with sanasol or 30 min before its administration. Method provides decreasing neurotoxicity based on antioxidant and immunostimulating properties of used preparations that are developed in the claimed administration regimen maximally.
EFFECT: improved method for decreasing neurotoxicity.
3 cl, 18 tbl
SUBSTANCE: group of inventions is provided concerning a method of treating a patient with cancer and envisaging administration of chloropromazine and pentamidine simultaneously or consecutively with interval within 14 days in quantities large enough to inhibit growth of the cancer, or in the form of pharmaceutical composition containing indicated agents in quantities, which jointly reduce proliferation of the indicated cancer cells. Pharmaceutical composition itself designed to indicated treatment and pharmaceutical kit are also disclosed. Proposed therapeutical agents separately do not show any antiproliferative effects.
EFFECT: achieved synergetic antiproliferative effect in absence of toxicity regarding normal cells.
17 cl, 1 tbl, 2 ex
FIELD: medicine, in particular oncology and gerontology.
SUBSTANCE: invention relates to foodstuff kit for weekly diet containing (g): dogrose flour 205-210; buckwheat grain 340-350; bran 65-70; green tea 32-35; coarse bread 980-1000; beans 190-200; lentil 190-200; fresh sheetake and mantaki mushrooms 290-300 and desiccated ones 145-150; fresh salmon 290-300; chicken 140-150; onion 190-200; garlic 90-95; carrot 1450-1500; tomato 1450-1500; white cabbage 450-500; sprout 450-500; cauliflower 450-500; broccoli 290-300; red beet 470-500; paprika (red, yellow, green) 340-350; celery 18-20; dill 18-20; parsley 18-20; sagebrush 18-20; bilberry 380-400; grape 380-400; pineapple 2600-2700; papaya 2400-2450; lemon 240-250; orange 260-270; olive oil 340-350. Claimed kit is useful in immunity reducing and increasing of chemotherapy effectiveness.
EFFECT: kit for prophylaxis and treatment of tumor diseases as well as for long-liver nutrition.
FIELD: organic chemistry, medicine, biochemistry, pharmacy.
SUBSTANCE: invention relates to benzamide derivatives possessing with inhibitory activity with respect to tyrosine kinase Flt-1-receptors VEGF that can be used in treatment of neoplastic disease. Invention describes a pharmaceutical substance comprising compounds of the group 2-[(4-pyridyl)methyl]-amino-N-[R1]-benzamide wherein R1 means 4-chlorophenyl, 4-methylphenyl, 4-chloro-3-(trifluoromethyl)phenyl or 3-(trifluoromethyl)phenyl possessing with the inhibitory activity with respect to tyrosine kinase Flt5-2-receptors VEGF associated with neoplastic disease and angiogenesis. Also, invention describes novel compounds of the group 2-[(nitrogen-containing heterocycle)methyl]-amino-N-[R1]-benzamide wherein nitrogen-containing heterocycle is represented by 4-pyrodyl, 4- or 5-quinolinyl, 2-imidazolyl, and a method for their synthesis. Also, invention describes a pharmaceutical composition comprising abovementioned compounds possessing the inhibitory activity with respect to tyrosine kinase VEGF receptors used in treatment of neoplastic disease.
EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.
17 cl, 2 tbl, 74 ex
FIELD: medicine, oncology.
SUBSTANCE: the present innovation deals with treating patients with generalized skin melanoma at single and multiple cerebral metastases and extracerebral lesions. The method includes systemic immunochemotherapy. For this purpose, on removing a single cerebral metastasis and melanomatous focus or at multiple cerebral lesions when operation is contraindicated it is necessary to carry out a 2-wk-long course of autohemoimmunochemotherapy, that is: 400 ml patient's autoblood should be sampled into a vial with hemoconservant and after sedimentation it should be divided into 2 equal fractions - plasma and autologous cell suspension (ACS). In separate vials it is necessary to prepare 3 media - 200 ml autoplasma (medium N1) and per 100 ml ACS (medium N2 and N3). One should incubate for 1 h in thermostat at 37° C medium N1 with 100 mg carmustine (mixture N1), medium N2 - with 25 mg metothrexate and 1 mg vincristine (mixture N2), medium N3 - with Reaferon at the dosage of 5×106 IU Reaferon (mixture N3). On the 1st d of the course one should inject mixtures intravenously by drops successively every mixture per 60 min, mixture N2 should be injected repeatedly on the 8th d of the course, mixture N3 should be injected by drops along paracetamol intake thrice weekly during the whole 2-wk-long course of therapy. Additionally, one should fulfill intravenous infusions of 150 mg carboplatin and 15 mg bleomycin upon 200 ml autoblood by drops on the 2nd and 4th d of the course, correspondingly. Totally, one should carry out 3-6 courses. The innovation provides stable regression of neurological symptoms due to decreasing the volume of cerebral metastasis and reactive perifocal cerebral edema and increasing relapse-free and metastasis-free periods and, also, life period in patients.
EFFECT: higher efficiency of therapy.
FIELD: medicine, biotechnology, pharmacy.
SUBSTANCE: invention proposes an antitumor peptide preparation representing human alpha-fetoprotein (AFP) fragment with amino acid sequence given in the description (fig. 1) and comprising 243 amino acid residues in primary structure and molecular mass 27 kDa. This AFP fragment is able for binding with AFP receptors, to inhibit estradiol-induced growth of hormone-dependent tumors, and to perform vector function of molecule in delivery of cytotoxic preparation into tumor cells. The preparation is obtained by culturing cells E. coli transformed with plasmid vector comprising cDNA gene of the third domain of human AFP. The peptide product is modified by additional methionine residue (Met) at N-end and by two amino acid residues (Leu-Glu) at C-end. Also, invention relates to the peptide preparation conjugate and cytotoxic agent, pharmaceutical composition and a method of treatment. As a cytotoxic agent the conjugate comprises a substance chosen from the following group: paclitaxel, docetaxel, doxorubicin, vinblastin, diphtherin, ricin and others. Method of treatment involves administration in the patient the conjugate or composition in the effective amount. Invention provides enhancing effectiveness and selectivity in treatment of hormone-dependent tumors.
EFFECT: enhanced effectiveness of treatment, valuable medicinal properties of preparation and pharmaceutical composition.
7 cl, 7 dwg, 7 ex
FIELD: pharmaceutical industry.
SUBSTANCE: invention relates to stone oil, extracted from stones of date (Phoenix pubeccens), yellow wood, or walnut containing specific amounts of triglycerides, diglycerides, monoglycerides, sitosterols, and cycloalanosterol. In one embodiment method for extraction of stone oil includes pressing of stones or powered stones, extraction with organic solvent or selectively by using of liquid in above-critical state to produce crude oil; discoloration thereof with adsorbing discoloration agent; dissolution of discolored oil in light ligroin; addition of stoichiometric amount of NaOH under stirring; settlement and aliquation; washing of organic phase with warm water to produce emulsion; addition of acetone to emulsion under stirring; layer separation and producing of oily phase in upper layer; treatment of oily phase to absorb thereof subsequently with neutral aluminum oxide and kaolin; removing after filtration of organic solvent from filtrate in nitrogen atmosphere; oily phase washing with warm water; heating of oily phase in nitrogen atmosphere to dehydrate thereof; and adsorption with neutral aluminum oxide. In another embodiment method for extraction of stone oil includes pressing of stones or powered stones, extraction with organic solvent or selectively by using of liquid in above-critical state to produce crude oil; stirring and heating of crude oil; adding of phosphorus acid to provide full degumming; addition of NaOH or Na2CO3 solution to degummed oil at the same temperature to provide full caustic purification; mixture settlement and aliquation to produce purified oil; washing of purified oil with pure water; addition of adsorbent to washed oil or heating thereof in vacuum to remove water and produce transparent dehydrated oil; discoloration thereof with adsorbing discoloration agent under heating in vacuum and under stirring in nitrogen atmosphere and oil heating up to certain temperature; passing of pure water steam at certain temperature and holding for certain time followed by stopping of purified water steam passing and nitrogen passing under stirring to remove moisture from oil. Pharmaceutical compositions improving of immunological function, increasing serum protein content and tumor growth inhibiting, which contains therapeutically effective amount of stone oil and one or more pharmaceutically acceptable adjuvants also are disclosed.
EFFECT: stone oil effectively improving of immunological function, increasing serum protein content and tumor growth inhibiting.
14 cl, 5 tbl, 11 ex
FIELD: medicine, oncology.
SUBSTANCE: method involves every day administration thiotriazoline on the background of doxorubicin treatment that is administrated once per a week for 4 weeks. Method provides carrying out correction of toxic damage of heart based on the complex anti-ischemic, membrane-stabilizing, antioxidant and immunomodulating effects of thiotriazoline. Invention can be used in correction of doxorubicin cardiomyopathy.
EFFECT: improved correction method.
3 tbl, 2 ex
FIELD: medicine; medical engineering.
SUBSTANCE: device has placental glycosaminglycans, glycerin, polyvinyl pyrrolidon, methyl cellulose, nipagin and purified water.
EFFECT: improved usability properties.
FIELD: medicine, gynecology.
SUBSTANCE: before operation it is necessary to sample patient's blood to divide it into plasma and blood cells. Blood plasma should be divided into three parts and frozen. After operation one should intravenously reinfuse by drops erythrocytic mass incubated for 30 min at 37° C with 2 g lendacyn followed by further lendacyn injection per 1g intramuscularly during the next 6 d. Blood plasma should be portionally defrosted, incubated at 37° C with: 10000 U trasilol, 0.15 g lisozyme, 0.02 g novocain, after incubation it is necessary to add 100 ml gelatinol to be reinfused for a patient on the 2nd, 4th and 6th d against the onset of therapy course through microirrigator withdrawn out of posterior arch during operation. The innovation provides combined impact of antibiotics upon pathological focus biotransformed with erythrocytic mass and local immunomodulating and resolving effect and, thus, decreased formation of adhesions, fistulas and cicatrices.
EFFECT: higher efficiency of therapy.
SUBSTANCE: method involves introducing recombinant human granulocytic colony-stimulating factor.
EFFECT: enhanced effectiveness of treatment; increased number of stem cells and their improved homing in lesion foci without polypragmasia effects.
FIELD: proteins, chemical-pharmaceutical industry.
SUBSTANCE: dry alpha-fetoprotein (AFP) is isolated from blood serum. Method involves fractionation of proteins with ethyl alcohol of ammonium sulfate, or sodium sulfate. After separation of supernatant the latter is dialyzed against sodium chloride solution and applied on affinity sorbent in sodium chloride solution with the concentration 0.1-1.0 mole/l and Triton X-100 with the concentration 0.01-0.5%. Sorbent is washed out with phosphate-saline buffer and AFP bound with sorbent is eluted by using glycine-HCl buffer solution. Eluate is neutralized and applied on Sepharose with antibodies raised to human IgG used as the second affinity sorbent followed by sterilizing filtration and lyophilization. Invention provides enhancing yield of AFP and its purity.
EFFECT: improved preparing method.
FIELD: medicine, biotechnology.
SUBSTANCE: invention relates to antibodies specifically binding to new human extracellular matrix polypeptides called as RGI; immunoconjugate containing the same and method for selective cell degradation; method for treatment of prostates cancer and metastasis in patients suffering from prostates cancer.
EFFECT: new method for treatment of prostates cancer.
28 cl, 7 ex, 7 dwg
FIELD: medicine, gastroenterology, pharmacology.
SUBSTANCE: invention relates to compositions comprising a substance possessing regulating activity, for example, inhibition of proliferation of cells expressing AILIM, for suppression of onset of intestine inflammatory diseases, in particular Crohn's disease, and colitis. Invention provides enhancing the therapeutic effect.
EFFECT: valuable medicinal properties of agents.
10 cl, 12 dwg, 2 ex
FIELD: medicine, oncology.
SUBSTANCE: method involves ligature of lung radix during the operation followed by taking off 400 ml of blood from pulmonary artery into capacity with glugicire, centrifugation at 1500 rev/min for 10 min and taking off plasma into another flask, and antibiotics are added into flask with remaining cellular suspension in the dose corresponding to the maximal daily dose, and neupogen is added to flask with plasma in the dose 60 mcg/kg of body mass. Both flasks are incubated at temperature 37°C for 30 min, and administrated in a patient by drops during the continuing operation. Method provides alignment of reduced resistance of body to infectious complications due to stimulating secretion of functional neutrophiles by bone marrow, elongation of time antibiotic residence in the body associated with formed blood elements and with absence of transient depression. Invention can be used in prophylaxis of suppurative-septic complications during the post-operative period in patients with the central lung cancer subjected for pneumoectomy operation.
EFFECT: improved method for prophylaxis.
FIELD: experimental medicine, oncology.
SUBSTANCE: the present innovation deals with increasing body resistance, at carrying out anti-tumor chemotherapy, as well. For this purpose one should sample about 0.7-1.0 ml venous blood, phylgrastim at the dosage of 10 mg/kg body weight should be incubated with this blood during 45 min at 37° C followed by reinfusion. The innovation provides total increase of body resistance due to stimulating the development of not neutrophilic leukocytes and monocytes only, but erythrocytes, as well, and decreasing the number of mast cells and reticular cells of stroma, and, moreover, it leads to decreased toxicity of anti-tumor chemopreparations.
EFFECT: higher efficiency.
1 ex, 1 tbl
FIELD: medicine, dermatology, in particular treatment of trophic ulcer (TU) and long-term open septic wound (LTOSW).
SUBSTANCE: claimed method includes application of wound-healthing composition onto TU and LTOSW in the first step of wound process. Said composition contains (mass %): methyluracil 0.9-1.1; pepsin 3.4-4.4; bentaine hydrochloride 3.6-4.4; sodium chloride 9.0-11.0; and balance up to 100,0: distilled water. In the second and third phases fine cut collagen preparations in form of 3-4 mm thickness layer are applied on purified TU and LTOSW surface. Then cotton carrier freely moistened with wound-healthing composition diluted with water in ratio of 1:2 is layered onto collagen layer. Multihole microirrigator and further sterile cotton carrier are sequentially applied over abovementioned cotton carrier.
EFFECT: effective treatment due to accelerated abruption of necrotic tissues, inhibition of microbial growth, decreased risk of infective, toxic and allergic affects, and improved tissue regeneration.
2 ex, 3 cl
SUBSTANCE: method involves introducing to a mammal therapeutic dose of drug having at least one subunit or at least one oligomer of mannan-binding lectin comprising one mannan-binding lectin subunit.
EFFECT: reduced risk of infection development; enhanced effectiveness in acting upon agents resistant to usual therapy.
43 cl, 2 dwg
FIELD: medicine, pediatrics.
SUBSTANCE: the present innovation deals with treating motor-autonomic disorders in children associated with affected function of central nervous system. For this purpose one should puncture perineural areas in the region of the main nervous trunks with alfetin dissolved in cerebrolysine. Additionally, one should puncture in projection area of cervical and lumbar spinal thickenings and areas that correspond to segmentary innervation of organs with affected function and, also, in scalp areas depending upon the character of patient's disorders. The method suggested provides improved autonomic-trophic impact of nervous system.
EFFECT: higher efficiency of therapy.