Antiishemic agent

FIELD: medicine.

SUBSTANCE: there is offered to apply 4-(2-hydroxyethyl)phenol(n-thyrozol) as a medicinal agent with anti-ischemic properties.

EFFECT: intravenous introduction of n-thyrozol improves the survival rate of animals with old myocardial ischemia and reperfusion, reduces a region of myocardial ischemia, and enables higher integrity of myocardial tissue.

3 tbl, 3 ex

 

The invention relates to medicine, specifically to pharmacology, and relates to agents that have anti-ischemic properties.

Known means exhibiting anti-ischemic properties: Trimetazidine [1, 2], Mildronate [2, 3], emoxipin [4], Mexidol [5].

The closest medicinal product (prototype) is Mexidol, has anti-ischemic and anti-oxidant properties [5-7].

Object of the invention is the expansion of the range of anti-ischemic funds.

The problem is solved by the use of 4-(2-hydroxyethyl)-phenol (n-tyrosol) as anti-ischemic tools.

It is known that n-tyrosol has antioxidant, hemorheological, antihypoxic and antiplatelet properties [8-13].

The use of n-tyrosol as anti-ischemic tools are not described in literature.

Brand new in the present invention is that as anti-ischemic tools used n-tyrosol. This property explicitly does not follow from the prior art in this field and it is not obvious to a person skilled. n-Tyrosol can be used for the treatment of patients with cardiovascular diseases.

Thus, this solution meets the patentability requirements of "novelty", "inventive step", "the industry is fair application".

Material and methods

Experiments for the study of anti-ischemic properties of n-tyrosol was carried out on 44 rats male Wistar weighing 200-250 g Animals were divided into 3 groups: 18 to rats intravenously injected with saline in equianharmonic quantities (control), 13 rats intravenously injected Mexidol in the dose of 10 mg/kg, 13 rats intravenously injected n-tyrosol in the dose of 20 mg/kg

Anti-ischemic properties of n-tyrosol and Mexidol was investigated using electrocardiographic (ECG) and morphological methods.

To create myocardial ischemia in anesthetized with methohexital sodium (100 mg/kg intraperitoneally) rats after inkubirovanija and connected to an artificial lung ventilation (ALV) HADES-2 put the ligature of the left coronary artery according to the method Accugane [14] without violating the topography of the heart in the chest. After 45 min the ligature was loosened, sutured the wound, placing the ends of the ligature under the skin. The ventilator was disconnected after restoration of spontaneous breathing; an endotracheal tube was removed after the onset of voluntary movements (reflex-turn). n-Tyrosol, Mexidol or equiano number of physiological solution (control) was injected intravenously 10 min of ischemia, after 4 and 22 h after the start of reperfusion.

After 25 hours after reperfusion rats was again narcoticyou, registrera the Ali ECG, put on a ventilator and tightened the ligature. To identify areas of hypoperfusion intravenous bolus was administered 0.2 ml of 5% solution of the dye patent blue violet (Sigma) and 10-20 extracted with heart. Areas of the myocardium with preserved perfusion was painted in green, repertoiremap remained unpainted.

Heart cross-sections with a thickness of 300 μm were prepared on a freezing microtome. The obtained slices (39-42 - depending on the size of the heart) were divided into 3 subject glasses, placing them on each one of three adjacent slices. Slices on one glass left in the native form (to assess areas of hypoperfusion), sliced on the other two slides were stained with nitrosonium tetrazolium (NBT) (Sigma) according to Seligman and Ruthenberg also finishing in [15] to identify the tissue dehydrogenase activity. Sections were concluded in glycerol-gelatin gel and scanned. Assessment of the extent of areas of hypoperfusion and changes in dehydrogenase activity was performed using Adobe Photoshop CS2.

Staining of sections of the PCT showed that in areas of myocardium subjected to 45 min of ischemia, there are areas with different degree of decrease in dehydrogenase activity; however, as a rule, areas with no activity and diminished activity of the mosaic was located amongst the areas with less significant change in enzyme activity. Observe the AMI uneven change in the activity of dehydrogenases in the area, subjected to a 45-minute ischemic effects, is a reflection of the high possibilities of collateral blood flow in rats [14]. Therefore, the area of infarction was conditionally accepted zone with a sharp decrease in dehydrogenase activity. For this purpose, sections of the myocardium was isolated areas, the total intensity of the color which was reduced by 2.5 times or more relatively intact areas of the myocardium.

During the whole experiment lasted ECG monitoring and the size of the zone of ischemic cardiomyocytes was evaluated at 25 h after reperfusion also on the EKG.

Statistical analysis was performed by standard methods using t-test, Student and criterion χ2using the software package Statistica for Windows 5.0".

The results are shown in examples 1-3.

Example 1. In the control group of animals within 1 day after the restoration of blood supply to the myocardium of the left ventricle died 7 rats of 18 (39%), death in all cases occurred in the first 2-3 h of reperfusion (table 1).

In this group of rats 1 day after the resumption of perfusion of the myocardium of the left ventricle on ECG were observed appearance of pathological Q wave, indicating the presence of necrotic changes in the myocardium, and a decrease in the amplitude of the T wave with respect to the initial values. The decrease of the voltage of subset in the control group was significant and caused by the presence of 5 animals from 11 ECG negative of the T wave (table 2). The presence in the subacute phase of myocardial infarction on ECG negative of the T wave characterized by the presence of zones of ischemic cardiomyocytes surrounding area of necrosis [16]. After 1 day after the restoration of blood supply to the myocardium in the control group remained significant decrease in the amplitude of R-wave relative to the original values (table 2)describing the violation process of depolarization of the ventricles due to the shutdown of part of the myocardium of the processes of excitation [16].

Table 1
The effect of Mexidol and n-tyrosol on the survival of animals after 1 day after ischemia-reperfusion
GroupThe total number ofSurvivors (%)
Control1811 (61%)
Mexidol1310 (77%)
n-Tyrosol1312 (92%)+
Note.+- p<0.05 compared with values in control group

Area of hypoperfusion is the control group took 32,1±2.2 percent. The area is sharply restricted dehydrogenase activity averaged 23,0±2.0% of the total area of the sections of the myocardium and took 72,7±5.1% of the area of the zone of hypoperfusion (table 3).

Example 2. In the group of rats with Mexidol has killed 3 rats out of 13 (23%) (table 1).

In this group of rats after 1 day after reperfusion of the Q wave on ECG was observed in 5 animals out of 10, which corresponds to the monitoring indicators. The average value of the T wave did not differ from control values, negative the T wave was observed in 4 animals out of 10. The decrease in R-wave in 1 day after reperfusion compared with baseline values were not reliable in nature (table 2).

td align="center"> to 266.8±22,2
Table 2
The effect of Mexidol and n-tyrosol on ECG parameters in rats after 1 day after ischemia-reperfusion
IndexControl (n=11)Mexidol (n=10)n-Tyrosol (n=12)
The original values1 dayThe original values1 dayThe original values1 day
HR, min-1463±9451±10444±9456±7478±13455±11
PQ, MS40,0±1,141,7±1,739,2±1,241,0±1,542,1±1,740,0±1,7
QT, MS85,0±2,681,7±5,087,5±2,279,8±3,681,7±3,378,3±3,3
Q µv024,3±9,1020,0±8,1031,0±13,4
R, µv302,3±39,4155,7±44,5#324,1±9,8233,0±52,7316,8±22,2to 266.8±38,8
T, µv256,3±28,85,6±22,2#EUR 236.9±27,120,6±34,6#105,6±33,4#+
Note.#- p<0.05 compared with baseline values;
+- p<0.05 compared with values in control group

Area of hypoperfusion in the group of animals treated with Mexidol, occupied 36,9±3.8% of the total area of cross sections of the myocardium. The area is sharply reduced dehydrogenase activity were 15.5±2.8% of the total area of the myocardium and 43.5±8.1% of the area of hypoperfusion and was 29% less than in the control group (table 3).

Table 3
The effect of Mexidol and n-tyrosol on the sizes of areas of hypoperfusion (in % of the total area of the sections and zones drastically reduced dehydrogenase activity (in % of the total area of the sections and in % of the area of hypoperfusion) after 1 day after ischemia-reperfusion
GroupArea of hypoperfusion, % of the area of infarctionThe area is sharply reduced dehydrogenase activity
percent of the area of infarction% of the area of hypoperfusion
31,2±2,223,0±2,072,7±5,1
Mexidol (n=10)36,9±2,315,5±2,8+43,5±8,1+
n-Tyrosol (n=12)31,9±2,316,7±1,8+53,7±6,3+
Note.+- p<0.05 compared with values in control group

Thus, intravenous administration of Mexidol has led to improved survival of animals undergoing myocardial ischemia with reperfusion. The use of Mexidol in rats during acute myocardial ischemia with subsequent reperfusion, according to ECG, reduce the area of myocardial ischemia and contributes to a higher compared to control preservation of myocardial tissue during ischemia and subsequent reperfusion.

Example 3. In the group of rats with the introduction of n-tyrosol died 1 animal out of 13 (8%) (table 1).

In the group of animals with the introduction of n-tyrosol in 1 day after reperfusion, the frequency of occurrence of the Q wave on ECG after an episode of ischemia-reperfusion injury in the experimental group was similar to the benchmark. The decrease in the amplitude of the T wave was less pronounced than in to the strole; the frequency of observations is the negative of the T wave in the group with the introduction of n-tyrosol was also significantly lower than the control values (1 of 12). In this group of rats revealed a clear trend (p<0,1) to normalize the amplitude of R-wave relative to the control group, and the decrease of this indicator in the context of the initial values after 1 day were not reliable in nature (table 2).

Area of hypoperfusion in the group of animals treated with n-tyrosol, occupied 31,9±2.3% of the total area of cross sections of the myocardium. The area is sharply reduced dehydrogenase activity was 16.7%±1.8% of the total area of the myocardium and 53.7±6.3% of the area of hypoperfusion and was 19% less than in the control group (table 3).

Thus, intravenous administration of n-tyrosol significantly increased the survival of animals undergoing myocardial ischemia with reperfusion, as compared with control, and with the group of animals treated with Mexidol. Introduction n-tyrosol rats during acute myocardial ischemia with subsequent reperfusion, according to ECG, reduce the area of myocardial ischemia and contributes to a higher compared to control preservation of myocardial tissue during ischemia and subsequent reperfusion.

Sources of information taken into account

1. Chekman I.S., Gorchakov N.A., Frantsuzova D.B, Mincer V.O. Cardioprotector - clinical-pharmacological TSA is you // Ukr. medicine honey. - 2003. - V.38, №6. - P.18-25.

2. Register of medicines of Russia. Encyclopedia of drugs. - M.: "radar-2007", 2006. - VIP. - P.573, 891.

3. Parkhomenko A.I. Metabolic approaches to the treatment of acute and chronic forms of ischemic heart disease and congestive heart failure // Ukr. practical use. doctor. - 1999. No. 1. - S-25.

4. Great Russian encyclopedia of medicines - M: Remedium, 2001. - S.

5. A.P. Golikov, main VP, Polumiskov V.Y., Fighters S.A., theological E.N. Experience in the application of cardioprotector Mexicor in patients with acute coronary syndrome // Diagnostics and treatment of disorders of regulation of the cardiovascular system: Sa. proceedings of the VI scientific-practical conference. - M, 2004. - S-492.

6. Lukyanova L.D., Atabaev R.E., Shepeleva HE Antihypoxic effects of some derivatives of 3-oksipiridina in isolated myocardium in rats // bul. the experimental. Biol. and the honey. - 1993. - T, No. 4. - S-368.

7. Kotlyarov A.A., Smirnov L.D. Effect of oxitetraciclina succinate on electrophysiological and hemodynamic parameters heart with thoracotomy and acute myocardial ischemia in the experiment /Experimental. and wedge, Pharmacol. - 2004. - C, No. 3. - P.14-17.

8. Mrs. N.V., Naruhina E.N., Krysin, A.P., a Pointer NM, Khrapova N.G., Burlakova E.B. ABOUT the relationship between structure and activity of natural and synthetic antioxidants // Cytology. - 1999. No. 9. - S-81.

9. Carpenter MB, Chernysheva GA, fungal V.I., M. Maslov, Cherkashina IV, Krysin, A.P., Sorokina I.V., Tolstikova T.G. Effect of n-tyrosol on blood viscosity and platelet aggregation, Byull. the experimental. Biol. and the honey. - 2007. - T, No. 1. - P.67-69.

10. Cherkashina IV, fungal VI, Chernysheva GA, carpenters could BE the Effect of n-tyrosol on thrombus formation in the rat model of arterial thrombosis // Chemistry and technology of vegetable materials: proc. Dokl. IV all-Russian scientific conference. - Syktyvkar, 2006. - s.

11. Carpenter MB, Chernysheva GA, fungal V.I., M. Maslov, Cherkashina IV, Krysin, A.P., Sorokina I.V., Tolstikova T.G. Effect of n-tyrosol on blood viscosity and platelet aggregation, Byull. the experimental. Biol. and the honey. - 2007. - T, No. 1. - P.67-69.

12. RF patent №2239423 "Hemorheological and antitrinitarian tool".

13. Fungal VI, carpenter MB, Chernysheva G.A., I. Golubeva, Krasnov EA Antihypoxic activity of n-tyrosol // Pharmacology - practical health: Mater. III Congress of pharmacologists Russia. Psychopharmacology and biological narcology. - 2007. - V.7, spec. vol. - S.

14. Kogan AKH Surgical simulation method coronacion myocardial infarction and aneurysm // Pathol. Fiziol. and experimental. therapy. - 1979. No. 3. - S-81.

15. Pier E. The Histochemistry. - M.: Publishing house of foreign literature, 1956. - 488 S.

16. Murashko CENTURIES, Strut the CENTURIES nski Electrocardiography. - M.: Medpress, 2000. - 312 S.

The use of 4-(2-hydroxyethyl)phenol as an anti-ischemic funds.



 

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