Hemostimulating medication

FIELD: chemistry.

SUBSTANCE: application of combination of diterpene alkaloids of aconitum baicalense: napelline, zongorine, mesaconitine, hipaconitine and N-oxide of epinapelline, obtained by extraction with chlorophorm as hemostimulating medication.

EFFECT: medication possesses expressed hemostimulating activity.

2 tbl, 2 ex

 

The invention relates to medicine, specifically to pharmacology and Hematology.

The high occurrence of anemic conditions and their influence on the duration and quality of life of patients [1, 2], is the basis of wide application gemostimuliruyuschee tools in medical practice.

Many gemostimuliruyuschee tools [1, 2].

The disadvantage of these tools is often their low efficiency and high frequency of adverse effects [3, 4].

Problem solved by the present invention is an expanding Arsenal of highly effective gemostimuliruyuschee funds.

The task is achieved by using the amount of diterpene alkaloids of Aconite Baikal as gemostimuliruyuschee tools.

New in the present invention is the use as gemostimuliruyuschee means amount of diterpene alkaloids of Aconite Baikal.

Used original means amount of diterpene alkaloids of Aconite Baikal was developed and received NII pharmacology" SB RAMS (Tomsk) in conjunction with the National research Irkutsk State Technical University (Irkutsk, Russia). Diterpenoid alkaloids were extracted from raw materials (herbs Aconite Baikal) in the form of free bases by extraction with chloroform, OTDELA is of n and identified according to standard methods [5].

In practical medicine for the treatment of anemic States, representing as independent disease, and pathological syndrome, occurring in various diseases, the most widespread means on the basis of recombinant forms of natural regulators of haematopoiesis (cytokines): erythropoietin, granulocyte colony-stimulating factor, granulocytemacrophage colonies-stimulating factor [1-3]. When this protein nature of the specified substance determines the existence of the only possible - parenteral route of its introduction into the body and a significant risk of side effects and complications [4, 5], due to the immunogenicity of these growth factors.

At the same time in NII pharmacology" SB RAMS was shown the principal possibility and high efficiency management functions of mesenchymal stem cells with the help of diterpene alkaloids of Aconite Baikal [6]. When oral detected marked stimulation of stromal progenitor cells of the skin, accompanied by accelerating the process of regeneration of the surface tissues [6].

While the effect of diterpene alkaloids of Aconite Baikal on the regeneration of hematopoietic tissue and the possibility of stimulation of hematopoiesis due to the activation of rodonachalnik the blood cells is not known. The experiment showed unexpected results.

The fact of the use amount of diterpene alkaloids of Aconite Baikal with achieving a new technical result consists in stimulation of hemopoiesis, for the specialist is not obvious.

The new properties are not derived explicitly from the prior art in this field and is not found in the patent and scientific literature.

The present invention can be used in medicine.

Based on the above, you should consider the claimed solution meets the criteria of "Novelty", "Inventive step", "Industrial applicability".

The experiments were conducted on 62 mice CBA, Animals obtained from the nursery of the Department of experimental biomedical modeling NII pharmacology" SB RAMS.

Studies were performed in accordance with good laboratory practice (GLP), the Order of healthcare of the Russian Federation No. 708n from 23.08.2010 "On approval of rules of good laboratory practice", the Federal Law of April 12, 2010 №61-FZ "On circulation of medicines", the "guidelines for the study gemostimuliruyuschee activity pharmacological substances" (2002) [7].

Example 1

Used the original tool was obtained from the herb Aconite Baikal. The aboveground part of the plants collected during the flowering period the Irkutsk region, were ground to particle size less than 5 mm, were treated with sodium carbonate solution and loaded into percolator. 2 hours after load filled him in 2000 ml of chloroform. Extraction with chloroform was conducted on a laboratory installation of continuous operation, consisting of percolator film evaporator and direct the fridge. Continuous circularity solvent is conducted at a speed of 2 l/hour Duration of exhaustive extraction was 5 days. The concentrated chloroform extract of percolator merged through the bottom valve of percolator and were concentrated in the evaporator. The obtained chloroform extract of alkaloidal was purified from alcaloide compounds by extraction with an aqueous solution of sulfuric acid. Further purification of the amount of alkaloids they were re-chloroform extraction from sulfuric acid extract after neutralization and its alkalizing to pH=9-10. To get rid of traces of alkali was washed with 50 ml water. Then the chloroform extract was filtered through a desiccant (15 g anhydrous sodium sulfate). At the last stage the received amount of alkaloids was evaporated to dryness. The product yield was 0.23%. Plant-derived agent, contained an amount of alkaloids: kapellen, somorin, mesaconitine, hypaconitine and N-oxide 12-epinepherine. Received alkaloids diluted distillirovanna the th water and brought them Concentratio to 0.0005%.

Example 2

The effectiveness of stimulation of hemopoiesis was determined on the model of cytotoxic mielosupression in accordance with the methodological guidelines for the study of gemostimuliruyuschee activity pharmacological substances [7].

Cytostatic myelosuppression was modeled by a single intraperitoneal injection of a solution of 5-fluorouracil (5-FU)1/2the maximum tolerated dose (MTD) (114 mg/kg). Starting from the next day after administration of cytostatic mouse experimental group received per os 0,0005% - aqueous solution the amount of alkaloids 0.05 mg/kg 1 time per day for 5 days. The control animals under similar conditions was injected solvent (distilled water) equivalent volume (0.2 ml).

5, 8 th and 12-th day after injection of cytostatic animals were killed by inhalation of CO2. From experimental and control mice by standard methods was determined by the content of reticulocytes in the peripheral blood, as well as indicators of bone marrow hematopoiesis (the total number of myelokaryocytes, myelogram) [8]. Content commiteeman precursor cells erythropoiesis (CFU-e) and stromal mechanical (CFU-f) in the bone marrow, as well as products eritropoeticescoe active substances (EPA) stick and non-stick items gemopoeticescoe microenvironment (SHM) was determined by culture is erodov in vitro [8].

Statistical data processing was performed by the method of variation statistics using t-student criterion. In case of deviation of the distribution in samples from a normal to evaluate the reliability of differences was applied nonparametric test of Wilcoxon signed-Mann-Whitney.

In the experiment, the introduction of 5-FU resulted in developing a deep and prolonged depression of erythroid germ blood. Throughout the timeframe of the study was to decline the content of eritrotsitov in hematopoietic tissue and the number of reticulocytes in the peripheral blood (up 10.2% 0,0% from the background on the 5th day, respectively). However, had expressed compensatory reaction from the pool of hematopoietic precursors. Registered the increase in the number of bone marrow CFU-e (table 1), associated with increased production of hematopoietic cells (EPA) stick and non-stick by myelokaryocytes. In addition, it was revealed increased functional activity of mesenchymal precursors (table 2), which plays an important role in the recovery of corrupted data cytotoxic agent SHM [9].

Introduction alkaloids led to a significant increase in the intensity of the recovery process of erythropoiesis. An increase in the content of eritrotsitov in the bone marrow, which is on the 8th with the t of the experiment was reached levels of background values and were 190,4% from the similar values in animals cytotoxic control (table 1). Reflection regenerate the hematopoietic tissue has been the emergence of a significant number of reticulocytes in the peripheral blood on the 5th day (35,63% of the background) and the increase in their number in the remaining observation period: up to 1151,0% and 237,7% of control for the 8-th and 12-th day, respectively.

Investigation of the mechanisms gemostimuliruyuschee actions alkaloidsetoposide drug revealed the dependence of the formation of the blood picture and bone marrow from the state of the pool parental bone marrow cells. The introduction of the solution of the pharmacological agents were accompanied by a sharp increase in the content of erythroid precursors in hematopoietic tissue on the 5th and 12th day experience (up to 132.6% and 347,6% of the cytotoxic control, respectively) (table 1).

The reaction of the haematopoietic precursors were largely determined by the change in the secretory functions of the stromal elements of the SHM. During the experiment were recorded increase production eritropoeticescoe active substances stick myelokaryocytes on the 5th, the 8th day of the study. At the same time, the alkaloids of Aconite Baikal does not affect the mobile fraction of the nuclears bone marrow. Production of EPA non-stick by surrounding cells remained at the level of the control values (table 2).

In addition, in the in-depth study of the influence of the investigated compounds the substances on the local mechanisms of regulation of erythropoiesis was installed another feature of the pharmacological action of alkaloids in respect of stromal progenitor cells from the bone marrow. The introduction of the drug on the background modeling cytotoxic mielosupression resulted in lower levels of fibroblastic SOME 5 day experience 1.7 times compared with the control group (table 2). Based on information in the literature [6] on the stimulating effect of the alkaloids of Aconite Baikal on mesenchymal precursors with predominant activation of processes of differentiation, it follows that the development of the identified phenomenon could be the result of the ripening acceleration data of the parental cells. Where compliance with the dynamics of changes of the studied parameter in such cytostatic control (table 2), when there is a significant part of CFU-f in regeneration SHM [9]. specifies the compensatory nature of drug induced shifts. Moreover, the development of the described reactions from mesenchymal precursors, probably served as the basis described above (stimulation of secretion EPA stick myelokaryocytes) enhance the role of the stromal component of the SHM in the restoration of suppressed 5-FU erythropoiesis due to the formation of de novo functionally active adherent of myelokaryocytes.

Thus, the results obtained indicate expressed gemostimuliruyuschee properties, the amount of diterpene alkaloids of Aconite Baikal in their oral use. This mechanismo the acceleration of regeneration of hematopoietic tissue is the activation of the parental hemo poetic cells by direct action on them alkaloids [6], and as a result of increased feeder capacity stromal compartment SHM. In addition, identified the importance of changes in the functional activity fibroblastic predecessors in the implementation gemostimuliruyuschee effects of the proposed tools.

Cited literature:

1. Metcalf D. Hemopoietic growth factors. 1. // The Lancet. - 1989. - Vol.15. - P.825-827.

2. Mashkovsky PPM Medicines: 15-ed. - M: PA "Publishing house New Wave", 2008. - 1206 S.

3. Volkova MA.. Shirin A.D. Erythropoietin in the treatment of anemia in cancer // Hematol. and transfusion. - 1997. - V.42. No. 6. - Pp.33-36.

4. Anderson J.A. Allergic reactions to drugs and biologic agents. JAMA. - 1992 - vol.268. - p.2845-2857

5. Pogodaev N.N., Gapova C., Vereshchagin A.L., Gorshkov A.G., Semenov A., study of the alkaloid composition of some species of Siberian aconito // Rast. resources, issue 2, 2000, S-84.

6. Sushkov GN,, Krapivin V., Nesterov Y.,, Povetieva T.N., Zhdanov V., Suslov NI, T. Fomin.. Udut E.V., Miroshnichenko, L.,, Kamanina E.V., A.A. Semenov, S. Kravtsova, Digi A.M. regenerative Mechanisms of action of diterpene alkaloids of Aconite Baikal // bul, experimental. Biol. and medicine. - 2012. No. 6. - S-827.

7. Digi A. M., Zhdanov V., Goldberg V.E. and other guidelines for the study gemostimuliruyuschee activity pharmacological substances // Bulletin of the scientific expertise and state control Le is arctonyx funds. - 2002. No. 1 (9). - P.29-32.

8. Goldberg ED, Digi A. M., Shah V.P. Methods of tissue culture in Hematology. - Tomsk: Publishing house of Tomsk state University, 1992. - 272 S.

9. Dygai A.M., Zyuz'kov G.N., Zhdanov V.V., et al. Effect of Transplantation of Peripheral Blood Mononuclears Obtained Using Granulocytic Colony-Stimulating Factor and Hyalu-ronidase on Hemopoietic Regeneration of Tissue during Myelosuppression // Bulletin of Experimental Biology and Medicine. - 2009. - Vol.148. No. 1. - P.120-125.

Table 1.
Dynamics of the content of erythroid cells in the bone marrow, reticulocytes in peripheral blood CFU-e in the bone marrow of mice CBA/CaLac after injection of distilled water (1) or the amount of alkaloids (2) against a single injection of 5-fluorouracil in1/2MTD, (X±m; R)
The timeframe of the study dayGroupErythroid cells
(×106/‰hip)
The reticulocytes (%)CFU-e (×105the nuclears)
before the introduction of1,77±0,3236,00±2,0522,00±3,02
510,18±0,05*020,31±0,08*12,83±2,7768.50±247*#
811,04±0,163,33±0,76*73,33±4,89*
21,98±0,30#38,33±1,26#40,00±to 7.61#
1210,59±0,08*14,33±0,49*20,33±2,91
20,81±0,21*34,00±1,03#70,67±7,52*#
Note: * - differences significant in relation to the background,
# - the difference is significant compared to the group treated with distilled water by infusion of 5-fluorouracil (1)

Table 2.
Dynamics of the level eritropoeticescoe activity in supernatant from non-stick, stick of kariolou bone marrow, the content of CFU-f in the coast is the second brain of mice CBA/CaLac after injection of distilled water (1) any amount alkalai Dov (2) against a single injection of 5-fluorouracil in 1/2MTD, (X±m; R)
The timeframe of the study dayGroupEPA from sticking to the myelokaryocytesEPA from the non-stick myelokaryocytesCFU-f (2.5×105the myelokaryocytes)
before the introduction of2,33±0,334,0±0,739,00±0,73
514,33±0,67*6,33±0,6716,00±1,98*
24,17±0,65*#13,00±1,90*#was 9.33±0,67#
814,33±1,099,00±0,587,17±0,79
24,50±0,9612,17±1,5*#8,17±1,01
1212,50±0,726,0±1,32*3,33±0,84*
2 2,17±0,409,51±1,69#1,83±0,48*
Note: * - differences significant in relation to the background,
# - the difference is significant compared to the group treated with distilled water by infusion of 5-fluorouracil (1)

The use amount of diterpene alkaloids herb Aconite Baikal: kapellen, somorin, mesaconitine, hypaconitine and N-oxide 12-epinepherine obtained by extraction with chloroform, as gemostimuliruyuschee funds.



 

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EFFECT: method allows to produce an extract with enhanced biological activity, nutritive value and bactericidal effect and to increase the extractive, flavouring and colouring substances yield.

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely physiotherapy. A method involves the administration of chophytol, diet therapy, magnetic therapy and exposure to electric current. The magnetic therapy is performed by a low-frequency sinusoidal alternating magnetic field. The exposure covers a collar zone and liver for 10-15 minutes. A continuous mode, frequency 50 Hz, grade I-II magentic induction are specified. The magnetic therapy is immediately followed by the exposure to complex-modulated pulse electric current with a neural-like pulse shape. The exposure is generated at the level ThVII - ThXI, paravertebrally to the right, and cover skin projections within the liver and gallbladder transversally. For this purpose, the electrodes are placed in 6 procedure fields: 1 - paravertebrally to the right at the level of the ThVII spine, 2 - paravertebrally to the right at the level of the ThXI spine, 3 - in a projection of the liver on a front surface of the body, 4 - in a projection of the liver on a back surface of the body, 5 - in a projection of the gallbladder on the front surface of the body, 6 - in a projection of the gallbladder on the back surface of the body. The 1st and 2nd electrodes are connected to the channel IV, while the 3rd and 4th electrodes - to the channel I, and the 5th and 6th electrodes are connected to the channel II. The channels I, II work in a frequency-drift group mode. The channel IV work in a continuous mode at fixed frequency 20 Hz. The current intensity is adjusted to apparent vibration sensation. Duration of a session is 10-15 minutes. The therapeutic course is 10-12 daily procedures.

EFFECT: method improves the motor function of the gall bladder ensured by the improved cholekinesis, hepatic circulation and cholestasis elimination.

2 ex, 3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a method for preparing an agent possessing anticoagulant, hypotensive and diuretic action. The method for preparing the agent possessing anticoagulant, hypotensive and diuretic action, consisting in the fact that milled dark mullein is extracted in 60% ethanol by re-percolation in a cluster of 3 unequally batched percolators; the prepared extract is settled, filtered, boiled down and dried in a vacuum drier in the certain environment.

EFFECT: agent prepared by the above method possess pronounced anticoagulant, hypotensive and diuretic action.

4 tbl

FIELD: medicine, oncology, amino acids.

SUBSTANCE: invention relates, in particular, to the development of an antitumor preparation based on natural substances. Invention relates to an amino acid preparation comprising at least one modified essential amino acid obtained by treatment of amino acid by ultraviolet radiation (UV) at wavelength 250-350 nm for 12-80 h at temperature 15-30oC or with ozone at temperature 15-25oC. The modified amino acid has no toxicity for health cells. Also, invention relates to a method for preparing such preparation. Invention provides the development of an antitumor preparation based on modified amino acids and expanded assortment of antitumor preparations being without cytotoxicity for normal cells.

EFFECT: valuable medicinal antitumor properties of preparation.

8 cl, 4 tbl, 2 dwg, 4 ex

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