Method of correction of biochemical and immunological disorders in conditions of experimental acute toxic hepatology, accompanied by an increase in immune responsiveness
The invention relates to medicine, namely to Hepatology and immunology, and the development of a method for correcting biochemical and immunological changes in conditions of experimental acute toxic Hepatology, accompanied by an increase in immune responsiveness. For this experimental animal parenteral (intraperitoneal) dissolved in physiological sodium chloride solution Tripeptide H2N-Gly-L-His-L-Lys-COOH in the dose of 1.5 mcg/kg of body weight ten times with an interval of 24 h between injections. The method provides a reduction of hyperactively immune system, which prevents the development of autoimmune process. table 2., 1 Il. The proposed method relates to medicine, namely to Hepatology and immunology, and can be used for the correction and prevention of biochemical and immunological changes in toxic hepatopathies.The reactivity of the immune system in various forms of liver disease plays a significant role in the pathogenesis of the disease, due to the direct influence of the immune system on the development of autoimmune complications of toxic and infectious lesions of the liver, on the transition of acute liver disease. // Wiss. Beitr. M. - Luther-Univ. Halle-Wittenberg R.-1987.- 99.-R. 192-198.), as well as in the maintenance of chronic pathologic process (yakhontova O. I., Karacharovo L. L. , Starkov S. A. Some immunological changes and their diagnostic value in chronic liver disease. // Braces. case. 1988 at M.V.Lomonosov.- 4.-S. 39-42. Many questions of the relations of the liver and immune system are solved in the experiment in the simulation of the pathological process in the liver by various hepatotropic agents (Alekseeva, I. N., Bryzgina T. M., P. C. I., Ilicevic N. C. the Liver and immunological reactivity. -Kiev: Nauk. Dumka, 1991.-S. 5). The reaction of the immune system depends not only on the damaging agent, but also on the dose, frequency and route of administration in the body. In terms of toxic hepatopathies can be expected as the reduction of the immunological reactivity (Molotkov A. O. impaired physiological mechanisms of regulation of the immune system in acute poisoning by organophosphorus compound karbofos: author for obtaining the academic degree of Cand. the honey. Sciences. - Saratov, 2002.-S. 20), and its increase (Hemp A. I. Endogenous immunomodulators as a factor in maintaining homeostasis in liver pathology: author. dis. for obtaining the academic degree of Dr. med. Sciences. - Kiev, 1989.-C. aryshire T. M., The P. S. I., Ilicevic N. In. Liver and immunological reactivity. -Kiev: Nauk. dum-ka, 1991. -S. 43-45). It can be effective only on condition of strict accounting stage of the disease, the nature and extent of immune disorders. Because in terms of toxic hepatopathies may experience different reactions of the immune system, shows the effectiveness of the treatment in some cases, immunostimulators, and other immunosuppressants. Immunostimulants effective against the immunodeficiency observed in some forms of liver disease, and to activate regenerative processes in the body. But the use of Immunostimulants (thymogen) in pathology of the liver it with increased reactivity of the immune system and a high level of immunoglobulins in the blood often leads to an exacerbation of the pathological process (B. I., Kuznik, V., Morozov, V. H. Khavinson. Cytomedines 25 years of experience in experimental and clinical studies. Saint-Petersburg.- "Science".-1998.-S. 197-198.). Immunosuppressants are shown in order to avoid the damaging action of immune mechanisms in the liver (Alekseeva, I. N., Bryzgina T. M. , P. C. I., Ilicevic N. In. Liver and immunological reactivity. - Kiev: Nauk. dum-ka,1991.-S. 43), as is ( ibid, S. 77).The closest correction method in the setting of acute hepatic toxicity CCL4(carbon tetrachloride (choral music theatre), another name for carbon tetrachloride (Chu), is the use of polysaccharide complex extracted from the roots and rhizomes of Filipendula castelpetroso (PSC SS), with immunosuppressive activity ( Mansimov O. C. abstract. Diss. for obtaining the academic degree of candidate of medical Sciences. Kursk, 1999, S. 12-16). In polysaccharide complex of the rhizomes and roots of Filipendula castelpetroso contains glucose - 13,6%, arabinose - 8,5%, xylose -%, galactose -%, mannose - 3,5%, ramnose, and 7.1%. In conditions of increased immunological reactivity caused by the introduction of the CCL4, PSC SS shows significant immunocorrective effect, that is, reduces the increased immunological reactivity to the level of healthy animals. This normalization of immunological reactivity is accompanied by decrease in the intensity of biochemical signs of liver problems: syndromes of cytolysis (the activity of alanine and aspartate aminotransferase), cholestasis (concentration of bilirubin, cholesterol) and hepatocarcinoma syndrome (fibrinogen concentration), indicating that the improvement in the liver. tis the development of autoimmune complications, the chronicity of the process and deterioration of the liver, and immune deficiency does not occur because of the reactivity of the immune system is reduced to the level of healthy animals, but not below it.The disadvantage of this method of correction is the complexity of obtaining UCS SS, the difficulty in standardization, since the percentage of active ingredients in the UCS SS may depend on the initial quality of the raw materials and conditions of the allocation, in addition, it is not clear what is the main active ingredient in this complex, which affects its effects from the exact percentage of monosaccharides or their sequence in the polysaccharides.The objective of the invention is to develop a way of correction of biochemical and immunological changes in the conditions of experimental acute toxic hepatopathies, accompanied by an increase in immune responsiveness.The objective of the invention is achieved by the fact that the experimental animal (rat) toxic hepatopathies involving high immunological reactivity, parenteral (intraperitoneal) dissolved in physiological solution (FR) sodium chloride suppressive immune response Tripeptide H2N-Gly-L-His-L-Lys-COOH (chemical nevala in the 24 hours between injections.In the developed correction method is used Tripeptide with weak immunosuppressive effect, and selected this dose and the multiplicity of its introduction that, without causing immunodeficiency, it limits hyperactively immune system. It can be argued that such immunotherapy positive effect on the liver, as it reduces the severity of biochemical symptoms of its destruction (syndrome of inflammation and cholestasis).This peptide isolated from blood plasma and is known for its stimulating effect on hair growth, collagen synthesis by fibroblasts, the accumulation of intercellular substance of connective tissue, which is largely due to the accelerated healing of wounds and ulcers under his influence, and why there are assumptions about the physiological role of this peptide as a stimulator of wound healing ( F. X. Maquart, Pickart L., Laurent M., Gillery P., Monboisse JC, Borel JP. Stimulation of collagen synthesis in fibroblast cultures by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+. // FEBS Lett. 1988. Vol. 238. N2. P. 343-346; Trachy R. E., Fors T. D., Pickart L., Uno H. The hair follicle-stimulating properties of peptide copper complexes. Results in C3H mice. // Ann. NY Acad.Sci. 1991. Vol. 642. P. 468-469.) Gly-His-Lys binds divalent copper and thus may participate in the activation of copper-containing enzymes, thereby providing haqour Century , Wegrowski J, Patt L. M., Trachy RE, Monboisse JC, Chastang F, Birembaut P, Gillery P. In vivo stimulation of connective tissue accumulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+. // J. Clin. Invest. 1993. Vol 92. N5. P. 2368-2376; Linder, M. C. Biochemistry of Copper. E. Freiden, Ed. Plenum Press. 1991).At the moment, because the known exact chemical formula of the peptide, it can be synthesized in any laboratory synthesis of peptides or purchased from companies involved in the delivery of biologically active compounds (Sigma, ICN, Merk, Bachim and others). We used the drug, synthesized in the Institute of chemistry, St. Petersburg State University.The drawing shows a histogram demonstrating immunocorrective effect of the peptide Gly-His-Lys, in terms of toxic liver injury by carbon tetrachloride.The method is as follows. As a model of acute toxic Hepatology (GTG), accompanied by stimulation of the immunological reactivity, we have chosen the model Hepatology, caused by carbon tetrachloride poisoning (choral music theatre), since this model has great similarity of the mechanisms of development of liver injury with clinical forms of acute, chronic Hepatology and liver cirrhosis of different etiologies in humans (Bluher A. F., Mayor A. J., Gerstein E. S. and other Patologii. /Ed. by A. F. of Bluger.-Riga.-1981.-Vol.9.-C. 5-24.). Experimental animals (rats), was injected intramuscularly 50% solution of the choral music theatre in vegetable oil at a dose of 3 ml/kg body weight for 5 consecutive days. Simultaneously with the arrival of hepatotropic poison, before and after him, ten times (once a day), intraperitoneally injected with 0.1 ml of a solution containing the peptide Gly-His-Lys in the dose of 1.5 mcg/kg of body weight, dissolved in sterile isotonic solution of sodium chloride. With such a multiplicity of the introduction of the peptide has a corrective action in respect of a number of biochemical indicators such as the level of beta-lipoproteins and severity of thymol samples, as well as weak IMMUNOSUPRESSIVE effect, which does not significantly weaken the immune system, prevents further increase the immunological reactivity that can lead to hyperactively and possible development of autoimmune complications. With the introduction of peptide in the same dose, but five normalizing biochemical changes, the effect is less pronounced, and increased in conditions of poisoning choral music theatre immunological reactivity is practically not reduced. Exceeding the recommended dosage more than 10 times (doses of 50 and 150 mg/kg) accompanied by a more pronounced suppressive aminotransferases, bilirubin concentrations, and beta-lipoproteins in the blood, as well as the severity of thymol samples, indicating the worsening of the functional activity of hepatocytes.Specific example Wistar Rats were divided into groups of 8-10 rats in each. Groups of animals are presented in table 1. The peptide Gly-His-Lys was introduced animals ten times (once a day) intraperitoneally at a dose of 1.5 mcg/kg and 100-fold greater dose of 150 mcg/kg GTG caused by five times (once a day) intramuscular injection of 50% solution of the choral music theatre in vegetable oil at a dose of 3 ml/kg of body weight, alternating legs. The peptide was administered prior to and simultaneously with the introduction of choral music theatre. The next day after the last injection of the choral music theatre rats were scored by exsanguination under ether anesthesia was given the serum, which was used for biochemical studies. In terms GTG observed increase in the activity of alanine and aspartate aminotransferase, bilirubin level, beta-lipoproteins and thymol turbidity of the sample, which is indicative of liver damage and the development of syndromes cytolysis, cholestasis and inflammation. In these conditions manifested corrective action of the peptide in a dose of 1.5 mcg/kg mass concentration of beta-lipoproteins and the level of thymol turbidity of the sample, and the whether the peptide at a dose of 150 µg/kg of body weight resulted in increased biochemical disorders (table 1).The same group of animals were taken to study the state of immunological reactivity. The intensity of the humoral immune response (GRI) to be judged by the number of antibody productive (KLA) and rosette (ROCK) cells in the spleen on day 5 after immunization of experimental animals (rats) (Mahlberg K. , Siegl, E. Immunological methods. Ed., Primes; TRANS. with it. A. P. Tarasova. - M., 1987-S. 273-282.). As the antigen used sheep erythrocytes (EB). The peptide was administered both before and after immunization according to the scheme in table.2 presents experimental Results on the drawing. The introduction of choral music theatre accompanied by the development GTG and stimulation of GMOs on the T-dependent antigen-EB. Introduction peptide under these conditions reduces the immunological reactivity that can be used for the purposes of immunological correction.Thus, the proposed Tripeptide in the dose of 1.5 mcg/kg tenfold, intraperitoneally experimental animals (rats, mice) significantly reduces the increased immunological reactivity, thereby inhibiting its hyperactively and the development of autoimmune complications, and also normalizes the number of biochemical parameters of blood serum, which indicates corrigiola the Denia has simultaneous corrective effect as regards the functional activity of hepatocytes, and in terms of immunological reactivity.
ClaimsMethod of correction of biochemical and immunological disorders in conditions of experimental acute toxic Hepatology, accompanied by an increase in immunological reactivity, characterized in that parenterally once a day for ten consecutive days is administered suppressive immune response Tripeptide H2N-Gly-L-His-L-Lys-COOH in the dose of 1.5 mcg/kg of body weight.
< / BR>where a is Gly; the remainder of the formula II
< / BR>m= 0 or 1; n= 2 or 3; R1and R2each independently of the other represents H, R1and R2both together represent also
< / BR>or
< / BR>where IS -(CO)-(CH2)q-(CO)rwhere q=1, 2, or 3, r=0 or 1, or-CO-CH=CH-CO-; X IS H, Cl or1-C6alkyl; and if the mean remains optically active amino acids and derivatives of amino acids, are included as D-and L-forms, and their salts, process for the preparation of compounds of formula I and their salts; pharmaceutical composition having the ability to inhibit integrin containing in its structure at least one compound of the formula I and/or one of its physiologically acceptable salts
FIELD: medicine, immunology, peptides.
SUBSTANCE: invention relates to a new composition of biologically active substances. Invention proposes the composition comprising of peptides of the formula: Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH wherein X means Gln and/or Glu; Y means Cys(acm) and/or Cys that elicits ability to inhibit the proliferative response for phytohemagglutinin, to induce the suppressive activity of mononuclear cells and ability of peptides to induce secretion of immunosuppressive cytokines of grouth-transforming factor-β1 and interleukin-10 (IL-10). The composition can be prepared by a simple procedure.
EFFECT: valuable biological properties of composition.
3 cl, 16 tbl, 9 ex