The product containing granulocyte colony-stimulating factor (g - csf) and tnf-binding protein
(57) Abstract:The invention relates to medicine, in particular to new drugs, containing immunoactive proteins, and is intended for the prevention and/or treatment of septic shock. The subject of the invention is a product containing G-CSF or its pharmaceutically acceptable salt and TNF-binding protein or its pharmaceutically acceptable salt. This product is designed for the treatment and/or prevention of septic shock. The invention expands the Arsenal of immunoactive protein funds. 12 C.p. f-crystals, 2 tab. The present invention relates to products containing granulocyte colony-stimulating factor (Granulocyte Colony Stimulating Factor = (G-CSF) or its pharmaceutically acceptable salt and the factor that causes necrosis of tumor cells (Tumor Necrosis Factor = TNF) binding protein (TNF-binding protein = TNF-BP) or its pharmaceutically acceptable salt in the form of a combined preparation, in particular for the prevention and/or treatment of septic shock.G-CSF is a major factor in the differentiation and maturation of granulocytes [Metcalf, D., Blood 67, 257-264 (1986)]. As was demonstrated that treatment with G - CSF leads to rapid improvement of the immune status of troparia [Gabrilove, J. L. et al., N. An Enol.J.Med. 318, 1414- 1422 (1988)] . Along with this, G-CSF confirmed the ability to reduce the impact of febrile seizures and nosocomial acquired infections in neutropenic patients and in cancer patients undergoing cytotoxic chemotherapy [Crawford, J. et al., N. Engl.J.Med. 325, 164-170 (1991)]. However, the effect of G-CSF on neutrophils is not limited to the differentiation and proliferation. It is obvious that it also plays an important role in regulating the functions of a Mature leukocytes [Lopez, A. F. et al., J. Immunol. 131, 2983 (1983)].Recently it was discovered that G-CSF reduces the fatality rate caused by lipopolysaccharide (LPS) septic shock in animals [Gorgen, I. et al., J. Immunol. 149, 918-924 (1992)]. It is also known in relation to soluble fragments of the TNF-receptor (TNF-receptor = TNFR) or chimeric polypeptides containing such soluble fragments [Lesslauer, W. et al., Eur. J. Immunol. 21, 2883-2886 (1991)]. Experimental animals with LPS-induced shock was widely used to identify compounds that could have potential in the prevention and treatment of shocks in humans. However, it appears that experimental animals caused by infection shocks are more correlated with the clinical situation than experimental animals with LPS-induced shock [Zanetti, G. et al., J. Immunol. 148, 1890-go fact shock observed distribution of peritonitis from Escherichia coli. At that time, as G-CSF and the aforementioned chimeric polypeptide alone showed a significant increase in the survival rate, it was found that, when G-CSF and chimeric polypeptide was administered together in the same state of the experimental animals was seen marked improvement in protection. Accordingly, the present invention is the preparation of products containing G-CSF or its pharmaceutically acceptable salt and TNF-BP or its pharmaceutically acceptable salt in the form of a combined preparation for simultaneous, separate or sequential use in the prevention and/or treatment of septic shock.The term G-CSF in the context of the present description and in the claims is used in its broadest sense, from the point of view of biological activity, understood by the person skilled in the art, and includes polypeptides (including natural or synthetic elements recombinant origin, modified or not), as defined and described (including their preparation and use in the scientific literature, for example, in any of the following patent publications: DE 3027105, EP 169566, EP 215126, EP 237545, EP 396158, EP 220520, EP 217404, EP 230980, EP 231819, DE 372 is described in EP 237545 [G-CSF broad-spectrum per person (hpG-CSF)], i.e. is preferred recombinant molecule may contain meinenemy residue at its N - end. Most preferred is G-CSF, having particular amino acid sequence which is encoded by the DNA sequences shown in EP 237545.In other words, the term G-CSF in addition to G-CSF natural origin contains any G-CSF-encoded DNA sequence which, upon expression in a prokaryotic or eukaryotic cell host leads to the production of a polypeptide product having at least a portion of the primary structure and one or more of the biological properties of naturally occurring hpG-CSF, as defined in EP 237545, with the specified DNA sequence chosen among:
(a) the DNA sequence shown in Table VII EP 237545, or its complementary chain;
(b) DNA sequences which hybridizing under any suitable conditions, hybridization, for example, as described in EP 237545 or described in "Molecular Cloning", Sambrook et al. 1989, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, and
(C) DNA sequences which, if not for the degeneracy of the genetic code, was hybridisable with the DNA sequences set is Telenesti.DNA sequences that hybridizing with the above sequences, the so-called mutant DNA sequences can be obtained using non-specific or siteprovides mutagenesis or by chemical synthesis or by using polymerase-cableway reaction (polymerase chain reaction = PCR) using primers based on the DNA sequences described in EP 237545, using methods known in this field and are described, for example, Sambrook et al. (see above), or in respect of PCR technology authors Innis et al. [PCR Protocols: A Guide to Methods and Applications, Academic Press, Inc. (1990)] . Thus, using mutant DNA sequence, can be obtained well-known in the field methods of mutant G - CSF, which are covered by the term "G-CSF", is described, for example, in the above patent publications. Mutant G-CSF characterized and their preparations are described in particular in EP 243153, WO 90/12874, WO 89/05824, EP 272703 or EP 456200.As indicated above, the term G-CSF includes G-CSF natural or recombinant origin, also in modified form, for example, when it is associated with chemical structural units, which, without changing the underlying biological actiity modification of polypeptides such as G-CSF, is a binding of water-soluble polymers, such as polyethylene glycols or polypropylenglycol, within the broad limits of molecular weight, for example, from 500 to 20,000 daltons. This leads to secure G-CSF, for example, polyethylene glycol, which can be practically non-immunogenic. In the prior art there are several options for linking polymer with G-CSF via different linkers, which are described in General, for example, in "Perspectives in Bioconjugate Chemistry", edt. by C. F. Meares, American Chemical Society, Washington 1993, and in particular, for example, in U.S. patent N 4179337. Modified G-CSF and their preparation are described, for example, in EP 401384, EP 335423 and EP 473268. Modified G-CSF also includes, for example, G-CSF, which demonstrates a different pattern of glycosylation, as is known, naturally occurring or recombinant G-CSF in the form of at least one additional polycarbohydrates circuit, as described, for example, in EP 370205.The term TNF-binding protein (TNF-BP) includes any protein or protein fragment, consisting of a sufficient number of amino acids that could be formed structure, which promotes the binding of TNF person regardless of where or how the same and any type of chimeric antibody to human TNF, as described, for example, in WO 91/02078. Such a chimeric antibody is an antibody in which the various parts of the molecule have different types of origin, in particular from human and animal, such as a mouse, rat or rabbit. In a preferred embodiment, such antibodies only complementary determining the plots have the origin is not from man, and if desired, additional amino acids in the variable regions. They can be prepared in accordance with methods known in this field, and as described, for example, in EP 239400 or in WO 90/07861.TNF-BP, as defined above, can be any naturally occurring or obtained by recombinant TNF-receptor or part thereof, preferably a protein, which contains part of the P55 or p75-TNFR person or is derived from it, and this part binds TNF, such as, for example, soluble forms of these receptors or, for example, so-called chimeric polypeptides that contain soluble portion of these receptors and all or part of the constant regions, but at least one constant region of the heavy or light chain of human immunoglobulin. Preferred such chimeric polypep the dimensional polypeptides are preferred, in which immunoglobulin part contains all of the region except the first region constant region of the heavy chain of human immunoglobulin, such as IgG, IgA, IgM or IgE, in particular, IgG, e.g., IgG1 or IgG3.Specialist in this field are very well aware of the fact that any amino acid of the immunoglobulin or part of the TNF - binding part can be removed or substituted by one or more amino acids, or one or more amino acids may be added, while TNF-binding portion binds TNF, and part of the immunoglobulin shows one or more of its characteristic properties. This is true for TNF-BP, as described above, without immunoglobulin part. Proteins TNF-BP, their isolation from natural sources or obtaining recombinant techniques, including the preparation of specific structures, such as, for example, in the form of a chimeric polypeptide containing, in addition to TNF - binding part and the immunoglobulin portion, as described, for example, in the following patent publications: EP 308378, EP 422339, GB 2218101, EP 393438, WO 90/13575, EP 398327, EP 412486, WO 91/03553, EP 418014, JP 127800/1991, EP 433900, US 5136021, GB 2246569, EP 464533, WO 92/01002, WO 92/13095, WO 92/16221, EP 512528, EP 526905, WO 93/07863, EP 568928, WO 93/21946, WO 93/19777 and EP 417563.This item is particularly the P55 TNFR, which binds TNF, or chimeric polypeptides containing such soluble part and the immunoglobulin portion, as defined above and as described in EP 417563. The person skilled in the art will understand that the determination of TNF-BP according to the present invention will also include TNF-BP, which has been modified by chemical means as described above in relation to G-CSF, for example, by binding a water-soluble polymer, such as polyethylene glycol or polypropylenglycol, according to the method described in the prior art, for example, in WO 92/16221.In addition, the present invention is the preparation of the above products, in which G-CSF and/or TNF-BP submitted/presented in the form of pharmaceutically acceptable (s) salt (s). For purposes of this description, the term "pharmaceutically acceptable salt" refers to both salts of the products of the present invention. Such salts of carboxyl groups can be formed by means known in the field, and include inorganic salts, for example sodium, calcium salts, ammonium salts, salts of trivalent iron or zinc, and the like, and salts with organic bases, as well as the salt formed, for example, with amines is, salts with mineral acids as hydrochloric acid or sulfuric acid, and salts with organic acid or sulfuric acid, and salts with organic acids as acetic acid and oxalic acid.Other objectives of the present invention is the use of G-CSF or its pharmaceutically acceptable salts in the preparation of medicines, in particular for the prophylaxis and/or treatment of septic shock patients receiving TNF-BP or its pharmaceutically acceptable salt as defined above, and the use of TNF-binding protein or its pharmaceutically acceptable salt as defined above in the preparation of medicines, in particular for the treatment and/or prevention of septic shock in patients receiving G-CSF or its pharmaceutically acceptable salt.In addition, the subject of the present invention is also a method of treating septic shock comprising introduction as TNF-BP or its pharmaceutically active salts and G-CSF or its pharmaceutically active salts.Products containing G-CSF and TNF-BP in the form of a combination, preferably in the form of a "kit of parts" containing these active ingredients in a separate opakowanie and/or treatment of septic shock. This means that the patient is injected with G-CSF and TNF-BP separately at the same time (simultaneously) or administered separately in a consistent way with synergistic effect. However, this combination also includes such combinations, in which G-CSF and TNF-BP is presented in the form of a mixture in the same package.For the prevention and/or treatment of septic shock using the above method G-CSF can be administered to the patient by any known method in the art, for example, intramuscularly, intravenously, subcutaneously, for example, using a subcutaneous implant, as described, for example, in EP 246322, orally in the form of a special oral dosing, as described, for example, in EP 459516 and EP 459795, pernaselli, as described, for example, in EP 565722, or the introduction into the lungs, as described, for example, in EP 505123.G-CSF may be made in the form of suitable dosage according to the specific method of administration. When G-CSF receive in the form of G-CSF-containing solution by any known methods, it is preferably kept in the solution at a temperature of about 4oC, but, depending on the type of the received G-CSF, it can also be stored in a frozen state. Alternatively, this solution can x the but for example, PEG-G-CSF in EP 335423. If desired, the G-CSF can be stored in the appropriate buffer with subsequent aseptic filtration through microporous filter or any other appropriate means in order to prepare suitable for injection of the drug.To produce dosage forms that are suitable for introduction to a patient a particular method to be applied according to the method of treatment according to the present invention, G-CSF may contain appropriate additives selected from known pharmaceutical carriers, fillers, diluents, stabilizers, for example, in the form of proteins, such as serum albumin human, protivoerozionnye substances, preservatives, soljubilizatory and emulsifiers, as they are described, for example, in DE 3723781, or presented in the form of stable hydrophobic composition, as described, for example, in EP 373679, or in the form of the drug in the form of particles with a slow release, as described, for example, in EP 263490 or EP 58481, or in respect of protected peg G-CSF, as described, for example, in EP 473268.The dose level and frequency of injection of G-CSF can be appropriately determined taking into account various factors, such as the severity ø be entered dosage, containing approximately 10-2000 mg/kg, preferably about 50-1500 mg/kg, most preferably about 10-250 μg G-CSF/kg body weight.TNF-BP according to the present invention are preferably administered parenterally by injecting, although there might also be other effective forms of introduction, as intra-articular injection, the formulations of oral action, transdermal iontophoresis or suppositories. One preferred carrier is physiological saline solution, but I believe that can also be used other pharmaceutically acceptable carriers. The primary solvent in the medium can be aqueous or non-aqueous nature. In addition, this carrier can contain other pharmaceutically acceptable excipients for modifying or maintaining the pH, osmolarity, viscosity, clarity, color, sterility, stability, rate of dissolution, or odor of the composition. Similarly, the carrier may contain still other pharmaceutically acceptable excipients for modifying or maintaining the stability, rate of dissolution, release, or absorption of a TNF inhibitor. Such excipients are those substances usually and customarily used for a composition which are prepared treatment composition, it can be stored in sterile containers in the form of a solution, suspension, gel, emulsion, solid, or digidratirovannogo or liofilizirovannogo powder. Such formulations may be stored either in a ready-to-use form or in a form requiring recovery immediately before use. The preferred storage of such compositions will be at temperatures not higher than 4oC and preferably at -70oC. it is Also advisable, to such compositions containing TNF-BP, stored and used at physiological pH value or close to it.In some embodiments of the invention, the introduction is used to retrieve a pre-selected range of concentrations of TNF-BP in the blood stream of the patient. I believe that maintaining concentrations of circulating TNF-BP exceeding 0.01 ng per ml of plasma is necessary for its effectiveness.The possible range of dosage for the treatment of septic shock may be in the range of from about 0.1 to 200 mg per kg of body weight of the patient for 24 hours, administered in equal doses in the range of about 4 to 15 times in 24 hours. The frequency of dosing and optimal dose will depend upon the pharmacokinetic parameters of TNF-BP used in the composition.Treatment of septic the following diagnosis of the possible septicemia. For example, treatment can begin immediately after surgery, or accident, or any other occasion when there is a risk of septic shock.Regardless of the method of introducing a specific dose is calculated according to the approximate body weight of the patient. Further refinement of the calculations required to determine the appropriate dosage for treatment involving each of the above compositions is performed in the usual way by an expert in the field of technology.Example
Septic shock caused by peritonitis from E. coli
Swiss mice albino (Jbm MoRo, weight 16-20 g) were administered by injecting an aqueous solution of G-CSF (Neupogen, Hoffmann-La Roche, Basle, CH; 0.1 mg/kg subcutaneously) and/or TNFR/IgG3 (TNF-BP, as described in EP 417563 consisting of the first 182 amino acids p55-TNFR person, merged with the hinge region of the heavy chain of IgG3 human rights and expressed in murine myeloma cells J558L; 2.5 mg/kg intraperitoneally), as shown in table 1. Peritonitis was induced by intraperitoneal injection of 106CFU (colony forming units) (colony-forming units = CFUs) culture of E. coli 25922 left on all night, which is about 500 times the number of organisms required to kill 50% of robotically from 5-10 mice each. The development of a common infection was stopped in all animals by administration of ceftriaxone (Hoffmann - La Roche, Basle, CH; 1 mg/kg subcutaneously) through 3.8 hours after injection of bacteria. This has led to bacterial cleansing power of the blood, but it was too late to prevent lethal septic shock in > 80% of the unprotected animals. Surviving mice received additional treatment with Ceftriaxone (ceftriaxone) 24 hours after infection. Animals that were seriously ill and were not able to eat and drink after 21 hours or longer after administration, wordplays and were recorded as adverse outcomes in therapy. For control took the blood out of the heart from at least one dead or userswindows mouse being treated groups and the control group was cultured on agar. Aggregated data from multiple experiments with G-CSF and mentioned specific TNF-BP, are presented in table 1.When used as the sole connection G-CSF resulted in a 20% survival value, and specific TNF-BP led to the magnitude of the survival rate of 22 %. These differences relative to the control group with saline solution with the magnitude of the survival rate of 10% is not very significant. A noticeable improvement in the defense was up to the sa and 2 hours before infection, in the scheme of admission to the combined form. All these animals showed visible signs of the beginning of septic shock after 3 hours after infection as the control animals. However, they did not die and was beginning to recover approximately 24 hours after infection. When combined scheme of introducing missed the last dose of G-CSF for 2 h before infection, the degree of protection was reduced to 65%. Counted the number of viable bacteria after 3.8 hours after infection, when the septicemia was stopped by injection of Ceftriaxone were only slightly lower in the group of G-CSF plus specific TNF-BP than in the group with saline solution (2.6 x 108CFU/ml as against 3.5 x 108CFU/ml, respectively). The content of endotoxin in G-CSF (25 µg/ml) and TNFR/IgG3 (250 μg/ml) was determined during limulus-test with the lysate of amoebocytes (sensitivity: to 0.06 endotoxin units/ml) (EU/ml), as is known in the art, it was < 0,05 endotoxin units/ml In accordance with this desensitization against lipopolysaccharides (LPS) can be excluded. The dependence on the dose of G-CSF in shock, caused by E. coli in Swiss mice albino, are shown in table 2 (G-CSF was administered in varying doses, as indicated, for 48 hours, 24 hours and 2 hours before paragenetically factor (G - CSF) or its pharmaceutically acceptable salt and the protein that binds the tumor necrosis factor (TNF-binding protein), or its pharmaceutically acceptable salt, and optionally, a pharmaceutically acceptable carrier.2. The product under item 1 for simultaneous, separate or sequential use in the treatment and/or prevention of septic shock.3. The product under item 1 or 2, in which TNF-binding protein contains the TNF-receptor or portion thereof, which binds TNF.4. Product according to any one of paragraphs.1 to 3, in which TNF-binding protein contains part of the P55 or P75 TNF receptor, and this part still binds TNF.5. The product under item 4, in which TNF-binding protein is a soluble portion of the P55 or P75 TNF receptors, soluble part of which still binds TNF.6. The product under item 4, in which TNF-binding protein is a chimeric polypeptide, which contains the soluble part of the P55 or P75 TNF receptor and all constant domains or part of a heavy or light chain of human immunoglobulin.7. The product under item 6, in which the chimeric polypeptide contains all domains except the first domain of the constant region of the heavy chain of IR represents JgG, in particular IgG1 or IgG3.9. Product PP. 1 and 2, in which TNF-binding protein is an antibody to TNF.10. Product according to any one of paragraphs.1 to 9, in which G - CSF is a natural G - CSF or the product of its prokaryotic or eukaryotic expression.11. The product under item 10, in which G - CSF is a product of prokaryotic expresii.12. The product under item 10 or 11, in which G - CSF modified.13. The product under item 12, in which G-CSF polietilenglikolya.
FIELD: medicine, oncology.
SUBSTANCE: the method deals with polychemotherapy and is fulfilled by the following technique: it is necessary to carry out blood sampling in these patients at the quantity of 25 ml: during the 1st d of polychemotherapy one should incubate 5 ml autoblood with 50 mg/sq. m metothrexate, during the 2nd, 3d, 4th and 5th d one should incubate per 5 ml autoblood with 15 mg/sq. m bleomycin daily in thermostat at 37 C for 30 min, carry out application anesthesia and introduce chemopreparations upon autoblood into endonasal tumor component or alternate intratumor introduction of chemopreparations upon autoblood at introducing them into maxillary antrum if it is affected with tumor process. The method provides local concentration and decreased toxicity of chemopreparations due to their intratumor introduction upon autoblood.
EFFECT: higher efficiency of therapy.
1 ex, 1 tbl
SUBSTANCE: the present innovation includes polychemotherapy and radiation therapy. Moreover, polychemotherapy should be carried out by the following scheme: on the 1st and the 8th d of the first and the third courses it is necessary to introduce doxorubicin, cyclophosphan, vincristine, and since the 1st to the 14th d - procarbazine and prednisolone; moreover, on the 1st and the 8th d of the second and the fourth courses one should introduce doxorubicin, bleomycin, vinblastine, dacarbazine. The method enables to decrease the quantity of late therapeutic complications, improves the results of relapse-free, total tumor-specific survival rate and decreases the number of polychemotherapeutic cycles.
EFFECT: higher efficiency of therapy.
FIELD: biotechnology, medicine, in particular treatment, prevention and diagnosis of diseases, associated with Neisseria meningitides.
SUBSTANCE: Claimed protein includes one or more N. meningitides protein fragments with known amino acid sequences, wherein said fragment contains one or more antigen determinants and has not more than 1977 amino acid from SEQ ID NO:1 and/or not more than 1531 amino acid from SEQ ID NO:2 which are described in specification with the proviso, that general protein sequence is not characterized by amino acid sequences represented in NO:1 and NO:2. Each claimed protein is encoded by nuclear acid (NA) with nucleotide sequence that defines protein amino acid sequence according to gene code. Peptides and nuclear acid of present invention are useful in drug production for treatment and prophylaxis of infections induced neisseria, as well as in production of diagnostic reagent for detection of neisseria or specific antibodies. Aldo disclosed is peptide- or NA-based pharmaceutical composition in effective amount with acceptable carrier. Said composition is useful for treatment of N. meningitides mediated infection. For prophylaxis composition is applied in vaccine form. Invention makes it possible to overcome diversity of neisseria antigen properties.
EFFECT: improved method for neisseria infection prophylaxis and treatment.
24 cl, 2 tbl
SUBSTANCE: invention discloses pharmaceutical compositions containing substance effective as modulators of biological activity "induced by activation of lymphocytic immunomodulating molecule (AILIM)" (known also as "induced common stimulator (ICOS)"), in particular modulating transduction of AILIM-mediated signal.
EFFECT: achieved suppression, treatment, or prevention of rejection of transplant arising in case of transplantation of organ, a part thereof or tissue.
11 cl, 7 dwg
FIELD: agriculture, animal science.
SUBSTANCE: one should inject thymogen intramuscularly about 60-45 d before calving thrice at 24-h-long interval at the dosage of 500 mcg/animal.
EFFECT: higher efficiency.
1 ex, 1 tbl
FIELD: medicine, biomolecular pharmacology.
SUBSTANCE: invention relates to the preparation that represents the product of biological and chemical treatment of vegetable biomass or its waste by using microorganisms. The parent raw represents carbon-containing biomass of vegetable origin wherein carbohydrate moiety represents 5% of the total biomass, not less. Biomass is mixed with water with addition of lactic acid microorganisms taken as a standard ferment with the parent content 0.2-0.5% per 1 l of an aqueous mixture. The mixture is incubated at 40°C for 6 h, not less, under condition of maintaining pH value at the level 6.75 ± 0.05 by addition of ammonium hydroxide. The prepared paste-like mass can be used as the assimilable protein precursor. Also, invention involves characterization of a method for preparing the preparation that involves treatment of the parent vegetable biomass with lactic acid microorganisms and fractional addition of ammonium hydroxide to provide pH value 6.75 ± 0.05. The final product represents the mass comprising the protein assimilable precursors. Invention provides the additional source of assimilable protein precursors.
EFFECT: improved preparing method, valuable properties of preparation.
6 cl, 7 ex
FIELD: veterinary ophthalmology.
SUBSTANCE: the present innovation deals with treating infectious-inflammatory ocular diseases in cattle due to complex treatment with applying antibiotic preparation and biolan, moreover, antibiotic should be introduced into parotid lymphatic center through catheter once or twice daily at the quantity of about 8-10 ml, and biolan - at the dosage of 3-10 mg into the lower conjunctival arch of an eyeball. Such a technique due to taking into account optimal conditions of intraocular liquid circulation provides synergistic effect of the preparations introduced.
EFFECT: higher efficiency of therapy.
1 cl, 1 ex
FIELD: medicine, otorhinolaryngology.
SUBSTANCE: the present innovation deals with preventing relapses of post-traumatic dacryocystitis. For this purpose, one should introduce a silicone tube perforated in checkered order along the whole length with openings of 1 mm diameter being closed at its free ends with stoppers into surgically developed lacrimo-nasal anastomosis to introduce in it 1.0 ml 1%-thymodepressin solution as immunodepressant once/5-7 d for 2 mo. The innovation provides suppression of excessive collagen-formation due to prolonged and steady impact of immunodepressant along lacrimo-nasal anastomosis.
EFFECT: higher efficiency of prophylaxis.
FIELD: medicine, oncology, pharmacy.
SUBSTANCE: after proving the pleural fluid sterility method involves its exfusion from pleural cavity and administration of antitumor chemopreparations. Firstly, 20 mg of bleomycetin is incubated with 20 ml of autopleural fluid, pleural fluid remained after exfusion is placed into packages "Gemakon" and centrifuged at 2000 rev/min for 60 min and liquid part is frozen. Pleural fluid is removed again as its accumulation and administration of incubated mixture of pleural fluid with bleomycetin is repeated also wherein the bleomycetin dose is increased up to 25 mg. In 2-3 days after removal of pleural liquid chemopreparations incubated with preliminary defrosted liquid part of pleural fluid are administrated each 5-7 days in the following sequence and doses: the first administration - 100 mg of cisplatin; the second administration - 100 mg of cisplatin; third, fourth and fifth administrations - 30 mg of doxorubicin and 1000 mg of cyclophosphan up to the total amount - cisplatin, 200 mg; doxorubicin, 90 mg, and cyclophosphan, 3000 mg. Method provides elimination of pleural fluid in full volume, to compensate loss of liquid, protein and trace elements, to reduce tumor size and to avoid toxic symptoms of chemotherapy. Invention can be used in the presence of exudative pleuritis in patients with lung cancer who can't to be subjected for operative and radiation treatment.
EFFECT: improved treatment method.
FIELD: medicine, biotechnology.
SUBSTANCE: invention relates to preparing biologically active substances from human and animal tissues. Invention proposes an antibacterial agent from pig and cattle platelets of molecular mass 0.5-15 kDa. Agent is prepared by freezing and defrosting platelet mass at temperature (-15)-(-20)°C for 24 h, centrifugation at 1000g, filtration of supernatant through dialysis membranes 15000 and 500 and elution of component comprising peptides by acetonitrile linear gradient from Sephadex G-50. Invention provides the synergetic antibacterial effect of peptides as component of the agent.
EFFECT: improved preparing method, valuable properties of agent.
8 cl, 6 tbl, 7 ex