Biomaterial and preparation with biomaterial, stimulating anti-tumour activity
SUBSTANCE: described is biomaterial, stimulating the anti-tumour activity, which contains lyophilisate of postnatal induced lymphatic nodes (PNILN) of a patient with cancer. Described is a preparation, stimulating the anti-tumour activity, which contains as an active substance lyophilisate of PNILN of the patient with cancer, and as a solvent, water for injections, cremofor RH-40, an emulsifier T-2, peach oil, glycerol, Kollidon CL-M and benzyl alcohol with the specified component ratio.
EFFECT: obtaining a novel preparation of domestic production, which contains biomaterial, stimulating the anti-tumour activity, possessing the expressed prolonged action, and convenient in application.
3 cl, 5 dwg, 3 ex
The proposed group of inventions relates to medicine, namely to Oncology, and can be used for biological therapy of tumors.
Modern advances in cellular and molecular biology have created real opportunities for active influence on antitumor immunity. It is assumed that the immune system can effectively combat the cancer cells, up to their complete elimination. Lymph nodes are one of the most important components of antitumor protection.
The known method of vaccine immunotherapy on the principle of reversing immunosuppression [patent US 7731945, 2010], which consists in the induction of the production of naive T cells, the effects on naive T cells to endogenous or exogenous antigens and the restoration of T-cell immunity by introducing the mixture of natural cytokines as adjuvant to endogenous or exogenous antigens cancer. Preferably the mixture of natural cytokines in combination with Timoshina 61.
Known vaccine therapy in patients with immunosuppression [patent US 6977072, 2005]. The method of immunization involves the activation of the regional lymph nodes, which consists of several stages: (1) perilymphatic the introduction of the mixture of natural cytokines. Some patients have a local accumulation of dendritic cells which have absorbed the antigen or entered into interaction with him, but you can't ripen. Perilymphatic introduction cytokines promotes the differentiation and maturation of dendritic cells;
(2) introduction to patient ciclofosfamida and nonsteroidal anti-inflammatory drugs. These drugs reduce T-cell suppression.
Known artificial lymph node [patent US 8101195, 2012]. The method of obtaining an artificial lymph node includes the following stages: (a) immunizing an animal using a cancer antigen and at least one adjuvant from the group consisting of complete adjuvant's adjuvant, CpG adjuvant, liposomes, BCG, polyI:C, R848 and LPS, (b) transplantation of material artificial lymph node, consisting of a polymeric biomaterial, including cytokineproducing stromal cells and dendritic cells for the immunized animal. Thus, it artificial lymph node, which causes the cancer antigen-specific immune response and enhances IFN production CDS-positive killer T cells or NK cells with cancer cytotoxic activity.
There is a method of treatment of disseminated cancer [patent US 8211425, 2012]. Immunotherapy is a method of treatment of a patient suffering from disseminated cancer, includes:
a) identifying in a patient one or more sentinel is limfaticeskih nodes and alternatively, the identification of all or part of the metastatic spread of cancer; b) resection of one or more nodes and, optionally all or part of metastases; C) allocation of metastat-reactive T-lymphocytes of these lymph nodes; d) in vitro multiplication of these metastat-reactive T-lymphocytes; d) the introduction of the thus obtained T-lymphocytes to the patient, in which T-lymphocytes are CD4+helper or CD8+T-lymphocytes.
The disadvantage of this method is the use of cells affected by a malignant tumor of the body that does not exclude the likelihood of progression and recurrence of cancer.
The closest analogue of the proposed biomaterial, stimulating antitumor activity, is an immunomodulator containing polypeptides in intact lymph nodes of animals representing the powder is yellowish-white in color. The method of obtaining the immunomodulator comprises homogenization of intact lymph nodes of animals in a solution of sodium chloride in the ratio of 1:3, autolysis, centrifugation, heating the supernatant to 80°C, cooling, removing the precipitate and drying the supernatant by freezing and premises in atmospheric freeze the plant with the subsequent final drying using infrared drying [patent RU 2257904, 2005].
The disadvantages of the data immunomodulator are:
Part immunomodulator, made of intact lymph nodes of animals, do not contain substances that are contained in the postnatal induced lymph nodes (NILU) or newly formed lymph nodes of cancer patients [Ghanaians SH New technologies of diagnostics and treatment of breast cancer / Creative surgery and Oncology, 2009. No. 1. - S. 6-9]), which have a higher biological activity [Gantsev S.K., K. Umezawa, Islamgulov DV, Husnutdinova E.K., Ishmuratova R.S., Frolova V.Y., Kzyrgalin S.R. The role of inflammatory chemokines in lymphoid neoorganogenesis in breast cancer / Biomedicine &Pharmacotherapy, 2013. -Vol. 67. No. 5. - R. 363-366]. When this method of obtaining a biomaterial used a solution of sodium chloride in the ratio of 1:3, autolysis at 4-6°C for 14 h, then heated supernatant to 80°C. due to the texture and physico-chemical properties of the composition is 0.15 M NaCl and the solution so obtained is poorly retained in the tissues, so that it does not provide prolonged action. In addition, this method of obtaining a biomaterial complex and time-consuming.
Object of the invention is the creation of a new biomaterial, stimulating antitumor activity.
Technical result - receiving material for tumor biotherapy, that reduce the likelihood of progression and recurrence of cancer, and tumor regression in some l is calization.
This technical result is achieved by the fact that the biomaterial, stimulating antitumor activity containing the lyophilisate lymph nodes mammal, according to the invention contains a lyophilisate postnatal induced lymph node cancer patient.
The invention is illustrated by the following figures: figure 1 shows NILU in breast cancer; the figure 2 is the same cross section; figure 3 - PILU formed with the stroma, stained with hematoxilin-eosin, ×60; figure 4 - profile of expression of inflammatory chemokines in PILU relatively intact lymph nodes; figure 5 - hierarchical analysis of the expression profile of inflammatory chemokines in PILU relatively intact.
Motivation applications are new data that were obtained in the result of long-term, multidisciplinary studies of the process of education NILU in cancer.
We have identified and studied in detail the formation NILU. These fabrics are reminiscent of the original intact lymphoid organs (figures 1, 2, 3), which ends after birth, with segregation in the area of T and b cells, dendritic cells, follicular dendrites cells, lymphatic vessels and large endothelial venules. When forming NILU activation of stromal cells leads to the release of molecules that control the processes of recovery, proliferation and survival of cells. One of the main roles in this process belongs to the interleukin-7 (IL-7R).
Creating a niche for incoming leukocytes occurs indirectly through cooperation with extracellular matrix components of adhesion molecules, cytokines and chemokines. The tumor necrosis factor (TNF) and lymphotoxin α (LT-a) expressed hematopoietic cells and play an important role in the activation of cytokines and chemokines in mesenchymal, stromal and endothelial cells, contributing to the creation of lymphoid niches [Francesca A., Ricardo P. Lymphoid neogenesis glue in chronic inflammatory diseases / Nature Rev. Immunol., 2006. - Vol. 6. - R. 205-217].
In transgenic mice, in which the observed overexpression of lymphoid chemokines or LT, is the development of organ-specific lymphoid organs (LO). It is known that increased expression of TNF-α in animals also leads to the formation of LO and chronic inflammatory reactions [Alitalo K., T. Tammela, T. Petrova Lymphangiogenesis in development and human disease / Nature, 2005. - Vol. 15. - P. 946-953; Bende R., van Maldegem f, van Noesel C. Chronic inflammatory disease, lymphoid tissue neogenesis glue and extranodal marginal zone B-cell lymphomas / Haematologica, 2009. - Vol. 94. No. 8. - P. 1109-1123].
Studies of chemokines and their receptors in mice led to the understanding of the molecular and cell-cell interactions, which underlie lymphoid organogenesis. With the development of lymphoid organs and maintaining microarchitecture whether vodnykh tissue cytokines inflammation, the family of TNF and LT-and closely interact with homeostatic chemokines CXCL13, CCL21, CCL19 and CXCL12. [Luther S., Bidgol A., Hargreaves, D., Schmidt, A., Xu Y., Paniyadi J., Paniyadi J., M. Matloubian, J. Cyster Differing activities of homeostatic chemokines CCL19, CCL21, and CXCL12 in lymphocyte and dendritic cell recruitment and lymphoid neogenesis glue / J. Immunol., 2002. - Vol. 169. - P. 424-433].
Important fact of the involvement of chemokines in specific metastasis of cancer cells (RK). It is found that breast cancer cells (breast cancer) human Express the receptor for chemokines CXCR4, whereas its complementary ligand CXCL12 is produced by the cells in tissues (bone marrow, lung and lymph nodes), which are often metastasize RK [Muller a, Homey C., Soto, N., Ge n, Catron d, Buchanan M.E. Ter-Minassian, T. McClanahan, E. Murphy, W. Yuan, S.N. Wagner, Barrera J.L., Mohar a, Verastegui e, Zlotnik A. Involvement of chemokine receptors in breast cancer metastasis / Nature, 2001. - Vol. 410. - R. 50-56].
The in vitro breast cancer cells are able to migrate toward CXCL12 gradient, and antibodies, neutralizing CXCR4, inhibit the formation of lymph node metastasis in experimental animals. Cells of lymphatic endothelium produce and other specific chemokines (CCL21), attracting to them the Republic of Kazakhstan from the primary tumor, which correlates with the presence of metastases in sentinel lymph node (LCS) [Takeuchi N., Fujimoto, A., Tanaka, M., Yamano T., Hsueh E, Hoon D. CCL21 chemokine regulates chemokine receptor CCR7 bearing malignant melanoma cells / Clin. Cancer Res., 2004. - Vol. 10. - R. 2351-2358].
The material for our study the Oia was the RNA samples, isolated from tissue of 15 lymph nodes of patients with clinically and cytologically proven diagnosis of breast cancer T2N1-3M0stage. Age of breast cancer patients ranged from 36 to 68 years (median 48 years). Total RNA was isolated from 200-300 mg of tissue histologically established normal lymph nodes and from NILU, Trizol reagent (Invitrogen, USA) according to standard Protocol. The quality of the selected RNA was determined by spectrophotometer NanoDrop 1000 (Thermo FS) the ratio of the optical densities of the A260/A280 (used samples only with a factor of 1.9 to 2.1). The selected RNA was treated with Dnazol. The expression profile of chemokines and their receptors was determined by fluorescent real-time PCR using a set of Human Chemokines&Receptors PCR Array (Qiagen) in two replications. The analysis of the expression of the following genes: chemokine (C-C motif) ligands: CCL1, CCL11, CCL13, CCL15, CCL16, CCL17, CCL18, CCL19, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8; chemokine (C-X-C motif) ligands: CXCL1, CXCL10, CXCL11, CXCL12, CXCL13, CXCL2, CXCL3, CXCL5, CXCL6, CXCL9; chemokine (C-C motif) receptors: CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR10, CCRLl, CCRL2; chemokine (C-X-C motif) receptors: CXCR5, CXCR3, CXCR4, CXCR6, CYFIP2; other chemokines and related genes: APLNR, BDNF, C5, C5AR1 GPR77), CCBP2, CKLF, CMTM1, CMTM2, CMTM3, CMTM4, CMKLR1, CSF3, CX3CL1, CX3CR1, TYMP, GDF5, GPR31, HCAR1, HIF1A, IL13, IL16, IL18, IL1A, IL4, IL8, CXCR1, LTB4R, MMP2, MMP7, MYD88, NFKB1, AIMP1, SDF2, SLIT2, TSR, TLR2, TLR4, TNF, TNFRSF1A, TNFSF14, TREM1, VHL, XCL1, XCR1.
Analysis of the gene expression, stroenie scatter charts and closterovirus was conducted using cloud applications based on the ΔΔCt analysis, on the website of the manufacturer of the test set [http://www.sabiosciences.com].
Our analysis of gene expression of cytokines and their receptors in the formation NILU revealed increased expression of genes CCL16, XCR1, CYFIP2, TNFSF14 and decrease in the expression of genes CXCL5 and CXCL12 (figure 4).
In our study, TNFRSF14 gene expression was increased in 4,82 times in PILU. The protein encoded by the gene TNFRSF14, is a member of a family of ligands of the TNF receptors. The protein of this gene activates two cell receptor - receptor cellular penetration of herpes virus (HVEM, TNFRSF14) and receptor lymphotoxin-β (LT-b) [Marsters, S.A., T.M. Ayres, Skubatch M, Gray C.L., Rothe M, Ashkenazi A. Herpesvirus entry mediator, a member of the tumor necrosis factor receptor (TNFR) family, interacts with members of the TNFR-associated factor family and activates the reduced factors NF-kappaB and AP-1 / J.Biol. Chem., 1997. - Vol.272. -No. 22. - R. 14029-14032]. Interaction TNFRSF14-RLT-b induces a cascade of reactions, mediated by transcription factors NF-KB (Re1A and Re1B), inducing the expression of genes involved in homeostasis, such as the genes for chemokines organizations tissue (CCL21, CXCL16 and others) and intercellular adhesion molecules (ICAM-1). TNFRSF14 together with its ligands-prologue, TNF, LT-a and LT-b, immunoglobulins, attenuator and T lymphocytes and CD160 forms a complex multisignal system regulation of inflammation and homeostasis of the immune system [Soroosh p, Doherty So, So So Herpesvirus entry mediator (TNFRSF14) regulates the persistence of T helper mmory cell populations / J. Exp. Med., 2011. - Vol.208. No. 4. - R. 797-809].
In autoimmune diseases characterized by chronic inflammation in the Genesis of which is attended by cytokines of the TNF family (TNFRSF14), there are typical events that lie in the penetration of antigen-specific T cells in the target organ, the involvement of other T cells, b cells and macrophages, and activation of cells that are in this place. The continuous production of cytokines TNF leads to the formation of organized lymphoid structures [ibid].
The analysis revealed a 30-fold increase in gene expression of chemokine CCL16 in PILU. Chemokinesis (C-C motif) ligand 16 (CCL16) is representative of a small family of CC chemokines, which are known under different names, including chemokine expressed in the liver (LEC) and Monotactin-1 (MTN-1). Chemokine CCL16 is expressed in the thymus, liver, spleen and stimulates the migration of monocytes and lymphocytes. Cellular expression of CCL16 in monocytes significantly induced by IL-10, IGF-γ and bacterial lipopolysaccharide. Gene CCL16 is located on chromosome 17, in the cluster of genes CC chemokines. CCL16 starts a cascade of reactions, interacting with the chemokine receptors on the cell surface, such as CCR1, CCR2, CCR5 and CCR8 [Nomiyama H., Hieshima, K., Nakayama T., Sakaguchi T. Human CC chemokine liver-expressed chemokine/CCL16 is a functional ligand for CCR1, CCR2 and CCR5, and constitutively exressed by hepatocytes / Int. Immunol., 2001. - Vol. 8. - P. 1021-1029]. In our study also revealed an increase of more than 4 times the expression of the chemokine XCL1 responsible for the migration of T-lymphocytes. Chemokine XCL1 belongs to a small family of chemokines, which retained only 2 of the 4 conservative cysteine, characteristic of all chemokines, for chemotaxis of T-cells. The gene is located along with the related chemokine XCL2 on the long arm of chromosome 1. Is expressed CD8+T cells and activated NK cells [Dorner B.G., Dorner M.V., X. Zhou, C. Opitz, R.A. Kroczek Selective expression of the chemokine receptor XCR1 on cross-presenting dendritic cells determines cooperation with CD8+T cells / Immunity, 2009. - Vol. 31. No. 5. - R. 823-833].
The detected increase in the CYFIP2 gene expression in PILU 4.7 times. CYFIP2 gene encodes a cytoplasmic protein that interacts with the gene product, fragile X chromosomes (FMR1), is expressed primarily in brain tissue, white blood cells and kidneys. Currently assumes the role of CYFIP2 in the regulation of apoptosis. It is believed that inducible oncosuppressor p53 protein CYFIP2 performs a proapoptotic function in RK [R.S. Jackson 2nd, Y.J. Cho, S. Stein, P. Liang CYFIP2, a direct p53 target that is leptomycin B sensitive / Cell Cycle, 2007. - Vol.6. No. l. - R-103].
Research a number of authors have shown that the chemokine receptors CCR7 and CXCR4 are expressed at high levels in breast cancer cells. While their respective ligands, secondary lymphoid chemokines (SLC/CCL21 and CXCL12 produced of limp the political endothelium in lymph nodes, bone marrow, lungs, liver, which are the predominant location of metastasis in breast cancer [Muller a, Homey C., Soto, N., Ge n, Catron d, Buchanan M.E., T. McClanahan, E. Murphy, W. Yuan, S.N. Wagner, Barrera J.L., Mohar a, Verastegui e, Zlotnik A. Involvement of chemokine receptors in breast cancer metastasis / Nature, 2001. - Vol. 410. - P. 50-56; GunnM., Tangemann K, Tam C, Cyster, J., Rosen, S., Williams L. A chemokine expressed in lymphoid high endothelial venules promotes the adhesion and chemotaxis of naive T lymphocytes, Proc. Natl. Acad. Sci. USA, 1998. - Vol.95. - P. 258-263]. Inhibition of the interaction of CXCL12/CXCR4 significantly reduces the metastasis of the Republic of Kazakhstan with breast cancer in the lymph nodes and lungs [ibid]. This type of molecular signals should be the most common mechanism of targeting of tumor cells to specific designated purpose and may represent a potential therapeutic target for the inhibition of progression of tumors to lethal diseases from metastases [Zlotnik A. Chemokines in neoplastic progression / Semin. Cancer Biol., 2004. - Vol.14. No. 3. - R. 181-185]. Our data showed a decrease in the expression of secondary lymphoid chemokines CXCL12 5.8 times and CXCL5 in 5,27 times in PILU (figure 5).
Perhaps the decrease in the expression of ligands for the chemokine receptors CXCR2 and CXCR4, are highly represented in the membrane RK, reduces the likelihood of migration of cells in PILU, preventing metastasis of a tumor.
Performed studies allowed us to propose the hypothesis that obtained from NILU biological is kind of substance can be used for the treatment of cancer, tertiary prevention of cancer.
In the regional lymphatic system in the area of the tumor through the lymph flow increased PILU [patent RU No. 2422914, 2011]. The method of obtaining the proposed biomaterial is homogenization NILU, its subsequent extraction and separation of the supernatant by centrifugation with subsequent lyophilization. Upon receipt of this biomaterial is not used cells derived from affected by a malignant tumor of the body. The biomaterial is a lyophilized extract NILU gray color with a yellow tinge.
Example 1. After excision of the cancer patient NILU, prepare the homogenate was diluted with 0,85-0,9% solution of sodium chloride in the ratio of 1:10. Extraction of the produce 24 hours at a temperature of 3±0.5°C and pH values of 7.0, centrifuged 10 minutes at 3000 rpm at a temperature of 1°C. the Obtained supernatant is poured into a container and lyophilizers. The resulting biomaterial is a lyophilized extract NILU gray color with a yellow tinge.
The closest analogue means with the biomaterial, stimulating antitumor activity, is effective for treatment of oncological diseases in the form of solution for injection in the following ratio of components (wt.%): 5-fluorouracil - 0,087, metronidazole - 0,5, water for injection, 1,81, low-molecular-weight polyethylene - 66,527, emulsifier T-2 - 16,778, Lutron F 127 (Lutrol F 127) -1,677, cremophor RH 40 (Cremofor RH 40) - 2,520, olive oil - 10,070 [patent RU 2445097, 2012].
Object of the invention is the expansion of the means to treat cancer.
The technical result - getting a new means of domestic production, containing the biomaterial, stimulating antitumor activity, is effective for injection route of administration with a uniform distribution in the tissues due to the composition of the injection solvent, pronounced with prolonged action, easy to use.
This technical result is achieved in that the means for encouraging antitumor activity, in the form of a solution for injection that contains the active substance and the solvent include water for injection, cremophor RH-40, emulsifier T-2 and oil, according to the invention the active substance and the solvent is taken in the ratio 1:2, as an active substance contains a lyophilisate postnatal induced lymph node cancer patient solvent additionally contains glycerin, kollidon CL-M and benzyl alcohol, and vegetable oil contains oil of peach in the following ratio, wt.%:
|Oil peach||at 10.64|
|Benzyl alcohol :||1|
|Water for injection||61,76|
Peach oil (GOST 21314-75) - vegetable oil extracted from peach kernels. Peach oil is used for the preparation of injection solutions of vitamins (ergocalciferol, retinol acetate), hormones (progesterone, sinistral, testosterone propionate, and others), camphor, krizanola and suspensions (biogenol).
Glycerin (FS 42-2202-84) - non-volatile co-solvent.
Cremophor RH-40 (Cremofor® RH-40) BASF (Germany) No. 013635/01-2002 - ethoxylated hydrogenated castor oil, improves the structure of the solvent increases the bioavailability of the incorporated active substances through the surface-active properties.
Emulsifier T-2 (TU 18-17-05-76) gives the mass a sufficient elasticity, used as a stabilizer, plasticizer and emulsifier, improves the structure is the solvent, increases the bioavailability of the incorporated active substances through the surface-active properties.
Kollidon CL-M Kollidon® CL-M, BASF (Germany) - white hygroscopic powder. The thickener. Non-toxic, is a fine powder with a low bulk density, and 90% of particles have a size less than 15 microns. This substance acts as a stabilizer suspensions in liquid dosage forms for oral administration. When introduced into the body kollidon promotes the resorption of the active substance from the injection site, which, in turn, increases the bioavailability of the composition, and also reduces the risk of local side effects. The increase in the content of kollidon above 5.5 wt.% leads to a strong increase of the viscosity and, consequently, deterioration of passing the composition through a needle of the syringe. The concentration of kollidon less than 0.1% of the composition is insufficient for stable complex formation.
Benzyl alcohol GOST 8751-72 - preservative, it is used for disinfection of oil solutions of preparations for intramuscular injection in pharmacology.
Example 2. The composition 100,0 emulsion solvent biomaterial: water for injection, 61,76; oil of peach - at 10.64; glycerol - at 10.64; cremophor RH-40 (Cremofor RH-40) - 5,32; emulsifier T-2 - 5,32; kollidon CL-M - 5,32; benzyl alcohol is 1.0.
Get injectable emulsion solvent as education is om.
In a water bath melt cremophor RH-40 (Cremofor RH-40), emulsifier T-2, kollidon CL-M at 70°C in varicellae Cup. To add alloy peach oil, glycerin and 5% water for injection. The aqueous phase emuleret. To the resulting structure add the remaining water for injection. The composition emuleret for 20 minutes. To the resulting mixture are added benzyl alcohol. The resulting composition is thoroughly mixed, filtered, filled into sterile vials under tested and sterilized at 100°C for 30 minutes.
A ready means of injection solvent biomaterial in the form of an emulsion is a homogeneous viscous mass of white with a yellowish tinge, with a density of 1.14 g/cm3pH is 6.2. The resulting solution was emulsion for injection has a liquid homogeneous viscous.
After excision of PILU from him prepare the homogenate was diluted with 0,85-0,9% solution of sodium chloride in the ratio of 1:10. Extraction of the produce 24 hours at a temperature of 3±0.5°C and pH values of 7.0, centrifuged 10 minutes at 3000 rpm at a temperature of 1°C. the Obtained supernatant is poured into the vessel lyophilizer and dissolved in the emulsion injection solvent in a ratio of 2:1 (emulsion: lyophilisate). Due to the consistency and physico-chemical properties of the composition obtained solution is securely retained in the tissues when any cconom the introduction, due to component solvent has a prolonged action of the active substance. The resulting solution injectable biomaterial convenient and hygienic to use.
In the available scientific and medical and patent literature found no information about the popularity of the proposed biomaterial, stimulating antitumor activity containing the lyophilisate PNILE a cancer patient, as well as products containing lyophilisate PNILE a cancer patient as an active agent and as a solvent water for injections, cremophor RH-40, emulsifier T-2, peach oil, glycerin, kollidon CL-M and benzyl alcohol. Thus, the claimed group of inventions meets the criterion of "novelty."
The research of the authors has revealed that the biomaterial, stimulating antitumor activity containing the lyophilisate PNILE a cancer patient, when, for example, injecting a solution containing the biomaterial as an active agent and as a solvent water for injections, cremophor RH-40, emulsifier T-2, peach oil, glycerin, kollidon CL-M and benzyl alcohol at a certain ratio of components, provides the inhibition of tumor growth and increased life expectancy. Thus, the claimed group of inventions meets the criterion of "subretinally level.
Receipt of the proposed biomaterial and tools with the biomaterial, stimulating antitumor activity, easily and reproducibly achieved using the specified technical result. Thus, the claimed group of inventions meets the criterion of "industrial applicability".
1. The biomaterial stimulating antitumor activity containing the lyophilisate lymph nodes of a mammal, characterized in that it contains lyophilized postnatal induced lymph node cancer patient.
2. The biomaterial according to claim 1, characterized in that to obtain hold homogenization postnatal induced lymph node cancer patient in a physiological solution at a ratio of 1:10, extraction, and separation of the supernatant by centrifugation, lyophilization supernatant, while extraction of the produce 24 hours at a temperature of 3±0.5°C and pH values of 7.0, centrifuged 10 min at 3000 rpm and a temperature of 1°C.
3. A means of encouraging antitumor activity, in the form of a solution for injection that contains the active substance and the solvent include water for injection, cremophor RH-40, emulsifier T-2, and vegetable oil, characterized in that the active substance and the solvent is taken in the ratio 1:2, as an active substance contains liof the lysate postnatal induced lymph node cancer patient
the solvent additionally contains glycerin, kollidon CL-M and benzyl alcohol, and vegetable oil contains oil of peach in the following ratio, wt.%:
|Oil peach||at 10.64|
|Benzyl alcohol :||1|
|Water for injection||61,76|
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to methods for reducing B-cell count or treating a disease or disorder related to pathological activity of B-cells. That is ensured by administering a therapeutically effective amount of CD37-specific binding molecule and a therapeutically effective amount of mTOR or PI3K inhibitor into an individual. There are also presented a composition and a kit for treating non-Hodgkin lymphoma.
EFFECT: group of inventions provides a synergetic effect in administering CD37-specific binding molecules (SMIP) in a combination with mTOR or PI3K inhibitors for treating or preventing the B-cell related hyperproliferative disease.
36 cl, 9 dwg, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to pharmaceutics and concerns irinotecan liposomes or its hydrochloride containing irinotecan or its hydrochloride, neutral phospholipid and cholesterol, wherein the weight ratio of cholesterol to neutral phospholipid makes 1:3-5, and a method for preparing them.
EFFECT: liposomes have higher stability.
15 cl, 3 dwg, 10 tbl, 10 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to peptide derivatives of general formula
their stereoisomers, mixtures and/or pharmaceutically acceptable salts, to methods for preparing them, to pharmaceutical compositions containing them, to using them for treating, preventing and/or diagnosing conditions, disorders and/or pathologies involving sstr1, sstr2, sstr3, sstr4 and/or sstr5 somatostatin receptor expression.
EFFECT: preparing the compositions for preventing and/or diagnosing the conditions, disorders and/or pathologies involving sstr1, sstr2, sstr3, sstr4 and/or sstr5 somatostatin receptor expression.
14 cl, 5 tbl, 33 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to compounds of formula 1.0:
where Q represents tetrahydropyridinyl ring substituted. R5, R1 are selected from: (1) pyridyl, substituted with substituent, selected from group, consisting of: -O-CH3, -O-C2H5, -O-CH(CH3)2, and -O-(CH2)2-O-CH3, R2 is selected from group, consisting of: -OCH3 and -SCH3; and R5 is selected from (a) substituted triazolylphenyl-, where triazolyl is substituted with one or two alkyl groups, selected from group, consisting of: -C1-C4alkyl, (b) substituted triazolylpheenyl-, wheretriazolyl is substituted on nitrogen atom with -C1-C4alkyl, (c) substituted triazolylphenyl-, where triazolyl is substituted on nitrogen atom with -C2alkylene-O-C1-C2alkyl, (d) substituted triazolylphenyl-, where triazolyl is substituted on nitrogen atom with -C2-C4alkylene-O-CH3, and (e) substituted triazolylphenyl-, where triazolyl is substituted on nitrogen atom with hydroxy-substituted -C1-C4alkyl, and where phenyl is optionally substituted with from 1 to 3 substituents, independently selected from group, consisting of halogen; and their pharmaceutically acceptable salts and solvates, which are claimed as ERK inhibitors.
EFFECT: obtaining pharmaceutically acceptable salts and solvates, claimed as ERK inhibitors.
15 cl, 2 tbl, 32 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of immunology, medicine and biotechnology. Claimed are versions of anti-EPHA2 antibodies. Claimed antibodies are bound with polypeptide, consisting of amino acids 426-534 in SEQ ID NO:8. Also described are hybridomes, which produce such antibodies, and pharmaceutical compositions and methods of application of said antibodies and compositions.
EFFECT: invention can be used in medicine.
74 cl, 14 dwg, 14 ex, 1 tbl
SUBSTANCE: invention relates to compounds or their pharmaceutically acceptable salts, where compound has formula 1-a, in which R1 and R3 are absent, m represents integer number from 1 to 2, n represents integer number from 1 to 3, A represents , B represents or , where X2 represents O or S, R4a is absent, R4b is selected from the group, consisting of: , , , , and ; Rk is selected from C1-6alkyl and C1-6halogenalkyl, L and E are such as given in i.1 of the invention formula; or compound is such as given in b) of i.1 of the invention formula. Invention also relates to pharmaceutical composition, which contains said compounds.
EFFECT: compounds by i1, possessing inhibiting activity with respect to anti-apoptosis protein Bcl-XL.
27 cl, 6 dwg, 2 tbl, 126 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to novel cyclic compounds of general formula which possess properties of CaSR modulator. In general formula I group represents cycloalkyl, which contains 4-7 carbon atoms, optionally substituted with one or several similar or different substituents, selected from R2, R3, R4 or R5; A represents 1-naphthyl; R1 represents methyl, ethyl or n-propyl, each of which is optionally substituted with one or several, similar or different substituents, selected from halogen and hydroxy; R2 and R3 represent hydrogen; R4 represents hydrogen, halogen, hydroxy or C1-6alkyl; each R5 represents independently one or several similar or different substituents, represented by hydrogen or C1-6alkyl; G represents -C(O)NH2, C3-8cycloalkyl, C1-6heterocycloalkyl, C1-6heterocycloalkenyl, C3-8cycloalkenyl, C6-14aryl, C1-10heteroaryl, C6-10arylamino, hydroxyaminocarbonyl, C6-10arylaminocarbonyl, C1-4aminocarbonyl, C1-6heterocycloalkylcarbonyl, C1-10heteroarylaminocarbonyl, C6-10arylsulfonylaminocarbonyl, C6-14aryloxy, or C1-4alkoxycarbonyl, where said substituents are optionally additionally substituted with one or several, similar or different substituents. Other values of radicals are given in the formula of invention.
EFFECT: compounds can be applied in treatment, relief or prevention of physiological disorders or diseases, associated with impairment of activity of CaSR, such as hyperparathyreosis, and other diseases.
23 cl, 9 dwg, 3 tbl, 315 ex
SUBSTANCE: invention relates to the application of a solid medicinal product, which is heated under the impact of an alternating magnetic field, for further therapeutic treatment after surgical ablation of tumours and cancerous ulcers. The medicinal product represents a surgical implant, presented in the form of a physiologically acceptable fabric, sponge or film. The medicinal product contains magnetic particles, which generate heat when excited by an impact of the alternating magnetic field, and in this way, heat the medicinal product.
EFFECT: invention ensures considerable improvement of further treatment after operation on cancerous tumour in comparison with chemotherapy.
21 cl, 14 ex
SUBSTANCE: method involves teletherapy combined with a local electromagnetic hyperthermia, chemotherapy with cisplatin and bleomycin, metronidazole as an ingredient of a composite mixture introduced through rectum. Teletherapy is conducted with using a continuous course of 3D and IMRT planning with photons 6-18 MeV covering the tumour and regional metastases in a single basic dose of 1.6 to 1.8 Gy daily 5 times a week, 27-30 fractions, in a total radiation dose of 48.0-48.6 Gy. At the same time, the tumour is exposed to teletherapy with using a simultaneous integrated boost in a single basic dose of 2.0 to 2.2 Gy, 27-30 fractions, in a total radiation dose of 59.4-60.0 Gy.
EFFECT: method enables reducing a radiation exposure, a teletherapeutic dose on the surrounding critical structure, saving the time and conducting a fewer sessions of teletherapy, ensuring lower intensity and the number of toxic reactions, the total length of treatment, eliminating early recurrences.
SUBSTANCE: blood prostate specific antigen is measured. Radiation therapy is carried out, and Zoladex 3.6 mg is administered every 28 days before, during and after radiation. If the blood PSA is lower than 1 ng/ml three months after the beginning of the Zoladex therapy, administering the preparation is continued for the following 6 months. Radiation therapy involves a split course in the regimen of the common radiation dose fractionation to the total radiation dose 64 Gy. If the blood PSA is found to be below 1 ng/ml only six months after the beginning of the Zoladex therapy, administering the preparation is continued for the following 9 months. Radiation therapy involves the split course in the regimen of the common radiation dose fractionation to the total radiation dose 70 Gy. If the blood PSA is above 1 ng/ml six months later, Zoladex is administered in a combination with Casodex 50 mg administered daily for the following 12 months. Radiation therapy involves the split course in the regimen of the common radiation dose hypofractionation to the total radiation dose 70 Gy.
EFFECT: higher effectiveness and optimisation of treatment, substantial reduction of a number of complications on the part of a wide range of a patient's body systems, and prolonged remission.
FIELD: food industry.
SUBSTANCE: invention relates to functional food products and represents a method for production of a non-aqueous health-improving product that is administrated in the form of drops and contains strain of lactic bacteria suspended in vegetable food oil; vegetable food oil is mixed with vitamin D; Lactobacillus rhamnosus GG lactic bacteria are suspended in the produced oil mixture.
EFFECT: invention ensures combination of beneficial effects of the proposed composition ingredients and easiness of the latter dosing during intake.
11 cl, 1 dwg, 1 ex
SUBSTANCE: invention discloses a pharmaceutical composition in the form of a solution, having nootropic and neuromodulating activity, characterised by that it contains N-carbamoylmethyl-4-phenyl-2-pyrrolidone as an agent, hydroxyethyl starch as an auxiliary substance and water for injection. The invention also relates to a method of producing such a pharmaceutical composition, which includes adding hydroxyethyl starch to the water for injection which is preheated to 85-90°C and mixing until complete dissolution, adding N-carbamoylmethyl-4-phenyl-2-pyrrolidone powder and mixing at temperature of 75-85°C until complete dissolution, adding water for injection to obtain the end volume of the solution and mixing once more, holding the obtained solution for 35-40 minutes at temperature of 75-85°C, cooling to 20-30°C, sterilising by filtering through filters with pore diameter of 0.22 mcm and packing in ampoules, which are further sterilised for 8 minutes at temperature of 120°C.
EFFECT: composition is effective, safe and can be used for a long period of time.
2 cl, 3 ex
SUBSTANCE: invention relates to a pharmaceutical composition of clear solution containing peptide, where the said peptide is H-Inp-D-Bal-D-Trp-Phe-Apc-NH2, acting as a ligand of receptor GHS, or its pharmaceutically acceptable salt. This peptide forms a depot in situ after subcutaneous or intramuscular injection to a subject. The pharmaceutical composition additionally comprises PEG, the said peptide is in the form of the pamoate salt, and the clear solution is an aqueous solution.
EFFECT: significant increase of the hormone as compared to the preparation of the acetate salt.
13 cl, 1 ex, 2 dwg, 5 tbl
SUBSTANCE: 5-10 Pletnev drops are administered orally twice a day for 60-90 days according to the following scheme: on the first day in the morning, Pletnev drops No. 5 and in the evening Pletnev drops No. 7, on the second day in the morning, Pletnev drops No. 30 and in the evening Pletnev drops No. 4, or No. 44, or No. 68; Pletnev drops No. 5 represent the wild strawberry leaves infusion; Pletnev drops No. 7 is the greater plantain leaves infusion; Pletnev drops No. 30 is the bilberry shoot infusion; Pletnev drops No. 4 is the wild strawberry leaves and foalfoot leaves infusion; Pletnev drops No. 44 is the wild strawberry leaves and wormwood herb infusion; Pletnev drops No.68 is the common yarrow herb and wild strawberry leaves infusion in 95% ethanol with taking substance 15-25 mg per 1 ml of the infusion.
EFFECT: invention provides high-effective immune therapy of viral hepatitis C with no side effects.
13 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to biopharmacology and represents a composition, a method and a set, which include a single-chain antibody scFv and a penetration-enhancing substance, selected from amino acid sequences, with a peptide being bound with a protective group with N-terminus, and the penetration-enhancing substance facilitates delivery of large macromolecules (i.e. larger than 10 kDa) through intercellular contacts.
EFFECT: group of inventions provides delivery of therapeutic antigen-binding polypeptides into CNS by intranasal introduction for treatment of neurological disorders.
8 cl, 14 dwg, 8 tbl, 4 ex
SUBSTANCE: invention relates to a concentrated acid component for bicarbonate hemodialysis. The acid component includes sodium (Na+) in an amount of 2450.0-4550.0 mEq/l, chlorine (Cl-) in an amount of 2453.5-4553.5 mEq/l, hydrogen H+ (hydrochloric acid), succinate and citrate, each in an amount of 3.5-98.0 mEq/l. The invention also relates to a diluted acid component for bicarbonate hemodialysis, which includes sodium chloride 70.0-130.0 mEq/l, hydrochloric acid 0.1-2.8 mEq/l, succinic acid 0.1-2.8 mEq/l, and citric acid 0.1-2.8 mEq/l. The invention also relates to a concentrate for preparation of the acid component, which contains hydrochloric acid in a liquid form, and all other components in a dry form, as well as to a method of obtaining the acid component, which includes dissolution of dry chemical reagents from the said concentrate in water and addition of liquid reagents.
EFFECT: invention ensures obtaining a solution for dialysis, applied in case of acute and chronic renal failure.
14 cl, 1 tbl, 3 ex
SUBSTANCE: pharmaceutical composition can additionally contain ethylenediaminotetraacetic acid, polyvinyl pyrrolidone, polyvinyl alcohol, a preserving agent specified in a group: Nipagin, Nipasol, benzoic acid, sodium benzoate, sorbic acid, benzalkonium chloride. As a body-forming base, the composition can contain distilled water, polyethylene oxide 400, polyethylene oxide 4000, polyethylene glycol, propylene glycol, a phosphate buffer, a borate buffer, an acetate-borate buffer depending on a dosage form.
EFFECT: high therapeutic effectiveness, prolonged corneal contact of the preparation which reduces the number of instillations, avoids a risk of side effects and provides good tolerance.
5 cl, 5 ex
SUBSTANCE: composition includes a bactericidal substance - catapol - in amount of 2.1-2.5 wt %, zosterin in amount of 1.1-5.0 wt % and distilled water.
EFFECT: providing a composition which stimulates a reparative process in external protective tissue, having anti-inflammatory and radioprotective action.
1 tbl, 1 ex
SUBSTANCE: pharmaceutical composition contains a combination of dalargin in an amount of 0.1-4 mg and vitamin specified in B1 (thiamine hydrochloride or thiamine bromide) or vitamin B6 (pyridoxine hydrochloride) or a mixture thereof in an amount of 25-100 mg.
EFFECT: composition manifests higher efficacy.
4 cl, 2 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to pharmaceutics and medicine, and presents a pharmaceutical composition for injections for acute forms of parkinsonian syndrome characterised by the fact that an active substance is presented by a therapeutically effective amount of N-(2-adamantyl)-hexamethylenimine hydrochloride (hymantan), and additive substances are presented by water for injections or a physiological solution. The invention also concerns a method for preparing the pharmaceutical composition.
EFFECT: pharmaceutical composition possesses storage stability, releases the active substance easily that provides its high bioavailability and efficacy.
6 cl, 15 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to chemical-pharmaceutical industry, and represents a composition containing polyethylene glycol 30 to 150 g, an ascorbic acid ingredient 3 to 20 g specified in ascorbic acid, ascorbic acid salt, or a mixture thereof, alkali or alkali earth sulphate, and at least one electrolyte specified in sodium chloride, potassium chloride, and sodium hydrocarbonate in one litre of an aqueous solution; it additionally contains flavouring agents and is effective in the colon cleansing for preparing for an endoscopy, particularly a colonoscopy. The invention contains a unit dose for preparing the colon cleansing solution, the colon cleansing solution, the method for preparing and using the same, and the kit for preparing and using the colon cleansing solution.
EFFECT: composition is safer than the standard colon cleansing compositions of sodium phosphate, and thereby may be used for the patients exposed to an elevated risk of using the composition of sodium phosphate, and is more tolerable than the standard compositions of PEG (polyethylene glycol) leading to a better compliance with the regimen and providing the effective use by outpatients.
48 cl, 6 ex, 29 tbl