Water-soluble composition, possessive properties of cardio-protector

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of medicine and chemical-pharmaceutical industry, in particular, to medication, applied in case of myocardial infarction and operations on heart in conditions of artificial blood circulation. Water-soluble composition for intravenous introduction, contains ingredients with the following component ratio in wt %: D-glucose 23.40-28.60; potassium chloride 0.14-0.16; potassium salt of L-asparaginic acid 0.81-0.99; semi-magnesium salt of L-asparagenic acid (magnesium L-aspartate) 0.72-0.88; human insulin genetically engineered (in IU/l) 54.00-66.00; dinitrosyl complex of iron (II) with glutathione 0.19-0.23; water for injections - remaining part, with solution pH 7.4±0.1 at 22°C.

EFFECT: application of composition ensures limitation of myocardial infraction size, recovery of metabolic condition of heart at reperfusion, reduces injury of membranes of post-ischemic cardiomyocytes with smaller haemodinamics disorders as compared with traditionally applied medications.

2 dwg, 7 tbl, 4 ex

 

The invention relates to the field of medicine and is intended for use as a medicine in myocardial infarction and heart surgery under extracorporeal circulation.

The main damaging factors of ischemia and reperfusion of the heart are changes in the energy provision of myocardial cells and the generation of reactive oxygen species (ROS)associated with impaired intracellular ion homeostasis. Their impact on itemizedoverlay the myocardium is able to completely block oxidative phosphorylation, to damage the structure of the mitochondrial and plasma membranes, to cause contraction of the myofibrils, to be the cause of the lack of recovery of blood flow and death of cardiomyocytes. These violations require the use during recovery in coronary blood flow correctors metabolism and antioxidants to restore the energy supply of cardiomyocytes and inactivation generated ROS [1]. Typically, this approach reduces irreversible myocardial damage, the occurrence of arrhythmias and reduces postischemic dysfunction of the heart [2]. Despite advances in modern pharmacology, mortality caused irreversible ischemic and reperfusion injury of the myocardium, in Russia and the developed world remains in the high. This is due primarily to the lack of effective drugs, while having the properties of metabolic protectors, stabilizers of membranes and vasodilators. Currently, attempts are being made to solve this problem through the use of offsets metabolism or donors of nitric oxide (NO).

The use of nitric oxide (NO) and its donors in cardiology due to the fact that NO is an endogenous product of the myocardium involved in the regulation of vascular tone, cardiac contractility, platelet aggregation, protection of cells from necrosis and apoptosis [3]. Most of the exogenous NO-donors, recommended for the treatment of myocardial ischemia and acute cardiac failure, have been used as vasodilators or inhibitors of platelet aggregation [4]. These include organic nitrates (nitroglycerin, ernits, nitrosorbid), heterocyclic amides (nicorandil) and complex compounds of iron with NO (nitroprusside). I believe that the most important mechanism of action of the most studied donor NO - nitroglycerin - is the reducing pre - and posterski on the heart, reducing the need for hospitalization of energy resulting from the expansion of peripheral blood vessels and deposition of blood in the veins [5]. There is evidence that nitroglycerin limits platelet aggregation [6]. Its ability to reduce RA the measures acute myocardial infarction (mi) and damage to membranes ischemic cardiomyocytes was confirmed experimentally [7]. These facts formed the basis for selection of nitroglycerin as an analogue of the developed cardioprotector. Despite prolonged use in cardiology, in the available literature there is no information about its effect on metabolic indices of ischemic heart. Well-documented negative effects - the rapid development of tolerance (the phenomenon with no clear explanation and correction method), side effects (headache and more dangerous - collapse), the inconvenience of its use and storage [8-10]. Positive effects of nitroglycerin (and long-acting nitrates) on the life prognosis of patients with acute or recently suffered THEM to demonstrate failed. Thus, in studies ISIS-4 (Fourth International Study of infarct dementia Survival) and GISSI-3 (Gruppo Italiano per lo Studio della Sopravvivenza Nell Infarto Miocardico) did not reveal significant positive effect of nitroglycerin on mortality and the incidence of complications in patients after acute THEM [11]. In the latter case the recommendations of the European society of cardiology its intravenous administration is not recommended for use in all patients with acute IM.

To reduce metabolic changes in the myocardium during ischemia and reperfusion applied protectors, which are composed of the natural metabolites of the heart - "metabolic cocktail" glucose-insulin-potassium (GIK), aspartate potassium and MAG is s (asparkam, Panangin), succinate derivatives (Mexicor), and synthetic inhibitors of the oxidation of free fatty acids (Trimetazidine, Mildronate). Their cardiotoxin action is based on the improvement of the energy state ischemic cardiomyocytes support higher levels of ATP and phosphocreatine (RKF) in conditions of ischemia, which provides better recovery of cardiac function when resuming circulation [2, 12-15]. Most of these drugs reduces the level of circulating in the blood free fatty acids (FFA) or reduce their consumption by the myocardium [12, 13]. This limits the toxic effect of FLC on itemizedoverlay myocardium associated with damaged membranes, which leads to arrhythmias and suppression of cardiac function [14-16]. In addition, metabolic protectors increase the capture of K+and prevent the overload of cardiomyocytes ions of Ca2+and Na+contribute to the transport of glucose into cells [15]. The result is a decrease in the excitability and conductivity of the myocardium during reperfusion, less development of reperfusion contraction, reduction of arrhythmias and myocardial infarction [17]. Based on these data as one of the prototypes developed cardioprotector was selected widely used drug is a mixture of the BOOM. The feasibility of using the BOOM b is La confirmed our earlier recovery of metabolic, morphological and functional parameters of the heart in models of ischemia and reperfusion in vitro and in vivo [18-20]. In clinical trials of various compositions BOOM in patients with acute THEY were also marked improvements in biochemical parameters of blood and cardiac function [12]. At the same time was identified lack of effectiveness in the clinical outcome of the disease - low limit THEM and reduce mortality of patients [21, 22]. These features of the BOOM, as well as other metabolic protectors due to the lack vasodilatation ability and antioxidant activity. In addition, some patients were noted discomfort in the epigastric region, nausea, allergic reactions, and headache [21]. This substantially limits the use of the BOOM in clinical practice.

The results of experimental and clinical studies suggest the need for the introduction of safer and more effective NO-donors and metabolic protectors in practice the treatment of acute coronary syndrome. In this regard, the present invention is the creation of a water-soluble cardioprotector possessing properties of metabolism regulator, stabilizer membranes and vasodilator. The problem is solved by the combination of iron (II)-organic complex oxide is zhota, namely, dinitrosyl iron complex with glutathione (dnci-GLn), with polonnoe biologically active medium containing energy substrates and insulin.

It is fundamentally important that a stable polymer matrix organoiron complexes with nitric oxide is solid-phase compounds, their use in the composition of potential drugs requires the development of water-soluble forms. The approach used to create the drug, was the use of biologically active medium for dissolving stabilized by dextran dnci-GLn, with cardioprotective properties. Its membership includes: d-glucose, potassium and magnesium salts of l-aspartic acid and insulin. The use of such an environment reduces the potential cytotoxicity of the complex. It is known that the result of excessive bioavailability of nitric oxide is the blocking of mitochondrial respiration and inhibition of the tricarboxylic acid cycle and the key enzymes of the glycolytic flux (phosphofructokinase and glyceraldehyde-3-phosphate dehydrogenase) [23]. Under conditions of energy deficit that can lead to irreversible lethal damage to myocardial cells, which is excluded by the joint application dnci-GLT and biologically active environment.

Created cardioprotector not only is em a water-soluble composition (pH 7.4), containing d-glucose, potassium chloride, potassium salt of 1-aspartic acid, polymagnet salt of l-aspartic acid, insulin human genetically engineered, dnci-GLn and water in the following ratio of ingredients (wt.%):

d-glucose23,40-28,60
potassium chloride0,14-0,16
potassium salt of l-aspartic
acid0,81-0,99
polomania salt of l-aspartic
acid0,72-0,88
insulin human genetically engineered
(IU/l)54,00-of 66.00
Dnci-GLnfor 0,19 0,23
water for injectionrest

The composition of the proposed cardioprotector original and is not described in the available literature. The combination of its components is not obvious and cannot be selected empirically. Bibliography patents on dinitroaniline the complexes of iron (II) with low-molecular weight thiol-containing ligands does not contain information on the subject of the invention is the development on the basis of NO-donors such chemical structure cardioprotector to reduce damage to the heart during ischemia and reperfusion.

The method of preparation of cardioprotector for experiments on animals.

Consists of three stages: 1) preparation of the environment with energy substrates, 2) preparation of dinitrosyl iron complex with glutathione, 3) obtaining water-soluble forms of cardioprotector with a given concentration dnci-GLT.

The cooking medium with d-glucose, K-Mg salts of l-aspartic acid and insulin. To 100 ml of distilled water is added to 26.0 g of d-glucose, to 150.6 mg of potassium chloride and stirred at low heat (37-40°C) until complete dissolution. To the resulting solution add 904,0 mg of the potassium salt of l-aspartic acid and 800,5 mg pollumassiivi salts of l-aspartic acid and dissolved while stirring at room temperature. the pH of the resulting clear mixture was adjusted 2N HCl to pH 7,4±0,1 at 25°C. the Final concentration of the components is in mm: d-glucose - 1200,0; potassium chloride - 20,2; potassium salt of l-aspartic acid - 44,0 and polomania salt of l-aspartic acid 23,0. The solution is injected suspension of human insulin to a concentration of 60 U/l and mix thoroughly.

Preparation of dinitrosyl iron complex with the restored glutathione. In 57 ml of distilled water, the solution is ut 5 g of glutathione, in 3 ml of distilled water dissolve 305 mg sodium nitrite. The solutions are mixed and incubated at 4°C for 10 min before the appearance of pink color due to the formation of S-nitrosoglutathione. 5% dextran solution in citrate-phosphate buffer is prepared by dissolving 50 g of dextran (30-40 kDa) and 9 g of NaCl in 400 ml of distilled water. Next, 60 ml of this solution add 30 ml of 100 mm phosphate-citrate buffer (pH of 7.4), and then 130 ml of distilled water. The final volume of 5% dextran solution in the buffer 220 ml. is added a solution of S-nitrosoglutathione and after stirring was adjusted pH to 7.4±0,1 30% NaOH solution. To the resulting mixture are added 20 ml of distilled water with dissolved 450 mg FeSO4×7H2O and 2.5 mg of sodium citrate. The mixture becomes orange color caused by the formation of dnci-GLT. The solution is filtered through a 5 μm Millipore filter and poured into ampoules of 0.5 ml to dryness in vacuum. Chemical reactions leading to the formation of dncg:

sodium nitrite+glutathione→nitrosoglutathione;

nitrosoglutathione+FeSO4+glutathione→dnci-PT+disulfide with glutathione.

One ampoule contains 22 mg of dry matter or 1.8 µmol dnci-Get in terms of iron (II) complex.

Obtaining water-soluble forms of the drug. The drug is prepared by dissolving dnci-GLT in medium with d-glucose, K-Mg salts of l-aspartic acid and insuli the Ohm, to create a dose of 1 µmol dncg/kg of body weight.

Methods of assessment of biological activity of cardioprotector.

To study the properties of the drug used model of regional ischemia and reperfusion in rats in vivo, providing an analysis of its impact on hemodynamics, limiting the size of the myocardial infarction, the recovery of the metabolic condition of the ischemic heart and reducing damage to the membrane of cardiomyocytes.

The model of regional ischemia and reperfusion in rats in vivo. In anesthetized with 20% urethane Wistar rats (120 mg/kg intraperitoneally) in terms of tracheotomy perform artificial ventilation of room air by the apparatus KTR-5 (Hugo Sacks Electronik). Jugular vein kateteriziruyut for staining of intact areas of the heart with a solution of Evans at the end of the experience. Carotid artery kateteriziruyut for registration of blood pressure (BP) using Mingograph-804 (Siemens Elema). The chest is opened by a longitudinal incision of the sternum and frees the heart from the pericardium. For the creation of regional myocardial ischemia left ventricular (LV) stitch atraumatic needle 5-0 under the left eye in the direction perpendicular to the major axis of the heart. While tightening the ligature on anterior descending coronary artery (PNA), are in the thick stitched attack, cease krovosnabzhayutsya myocardium; the weakening of ligatures leads to the restoration of coronary flow. Indicators of acid-base balance of arterial blood (pH, pO2and pCO2) during the entire experience is controlled by acid-base analyzer ABL-30 (Radiometer) and support at the physiological level. After preparation should be a 30-minute period of stabilization of hemodynamic parameters (initial state). Then perform the occlusion of the PNA for 40 min, the duration of reperfusion 60 min [20].

The sizing of myocardial infarction. At the end of the experience to identify areas of risk (WR) and the intact region of the myocardium occluder PNA and jugular vein injected bolus of 2% solution of Evans (2 ml). Then cut out the heart and separates the left ventricle, which is frozen at -20°C until histochemical sizing THEM colouring fabric 2,3,5-triphenyltetrazolium chloride. From LV prepared cross sections and thickness of approximately 1.5-2 mm, which is incubated for 10 min in 1% solution of 2,3,5-triphenyltetrazolium chloride in 0.1 M phosphate buffer (pH of 7.4 at 37°C). The samples are scanned, square and LA determined by computer planimetry using Imagecal. Then the slices are weighed to determine the mass of the left ventricle. In each group, calculate relations SP/LV mass and/WR in percent.

Meta is alicence the state of the heart. At the end of the periods of stabilization of hemodynamics (initial state), ischemia and reperfusion tissue WR frozen nippers of Wollenberger, chilled in liquid nitrogen for subsequent determination of metabolites. Frozen samples of myocardial tissue homogenized in cold 6% HClO4(10 ml/g of tissue) using a homogenizer (Ultra-Turrax T-25 (IKA-Labortechnik, Germany). Proteins precipitated by centrifugation at 3000 g and 4°C for 10 min Supernatant neutralize the 5 M K2CO3to a pH of 7.4. Sediment KClO4separated by centrifugation in the same conditions. Protein-free extracts stored at -20°C until determination of metabolites. Dry weight of the samples is determined by weighing a piece of tissue after extraction HClO4and drying at 110°C over night. Metabolites in tissue extracts determined spectrophotometrically, their contents are expressed in µmol/g dry weight.

Damage to cell membranes. Appreciate the increased activity of lactate dehydrogenase (LDH) and the MB fraction of creatine phosphokinase (MB-ck) in the plasma at the end of reperfusion. Blood is collected in heparinized tubes from the venous catheter in the initial state (before occlusion PNA) and after an hour of reperfusion. Plasma was separated by centrifugation (2000 rpm for 10 min at 10°C). The activity of enzymes in freshly isolated plasma determine the spectrophotometrically at λ=340 nm, using kits BioSystems and reagents company Sigma-Aldrich.

The indicators of the functional hemodynamics. Changes in average blood pressure (BP cf) and frequency of heart rate (HR) register on-line using Mingograph-804 (Siemens Elema) during the entire experience. The obtained results are statistically analyzed, comparing with the original and change in control (no drug) or under the influence of drugs comparison. Differences between groups assessed using analysis of varians (ANOVA) and considered statistically significant at p<0,05.

The Comparators. Used nitroglycerin (Perlinganit) and a mixture of glucose-insulin-potassium (GIK) of the following composition: glucose 2.8 M, insulin human genetically engineered 100 IU/l, KCl In 0.1 M Nitroglycerin, as a NO donor, provides peripheral vasodilatation and has koronarorasshiryayuschee effect, can reduce irreversible damage postischemic cardiomyocytes. GEEK metabolic protector, affects the processes of intracellular metabolism, limits the size of THEM[12, 20, 5, 7]. Currently, both drugs are used in clinical practice in the treatment of heart lesions in acute coronary syndrome.

Example 1. Limit the size of myocardial infarction.

Testing cardioprotector and compare its effects with the action is of nitroglycerin and BOOM carried out on the model of regional ischemia and reperfusion in rats in vivo. Performed two series of experiments with intravenous preparations: 1 - to occlusion of the PNA and 2 - after regional ischemia in the first minute of reperfusion. BOOM, nitroglycerin and cardioprotector was injected intravenously at a rate of 1 ml/kg of body weight of the animal. Dose of nitroglycerin (1 µmol/kg rat) was equimolar content dnci-Get in cardioprotector. In the control intravenously was administered an equal volume of saline.

Table 1
The impact of intravenous cardioprotector and Comparators on the size of myocardial infarction in rats in vivo
The route of administration of medicationThe infarction area/risk %
Control (n=22)before occlusion of the PNA or the first minutes of reperfusion47,3±2,1
Cardioprotector (n=12)32,4±2,3and
Nitroglycerin (n=12)before occlusion of the PNA37,7±2,7and
GEEK (n=12)39,2±2,2AB
Cardioprotector (n=12)29,5±2,0and
Nitroglycerin (n=10)in the first minutes of reperfusion40,3±2,9b
GEEK (n=10)36,8±2,1AB
Data are presented as M±M. Control - introduction of saline. Significantly different (p<0,05) from:andcontrol;bcardioprotector.

Histochemical analysis of the slices of the left ventricle after reperfusion revealed no significant differences in the amount of SP between groups. The average ratio LA/LV weight (in %) for all groups was 35,9±4.0 percent. This indicated the validity of the comparison data to limit THEM in the groups studied.

With the introduction of drugs before coronary artery occlusion was observed a significant decrease in comparison with the control. The ability to restrict THEM fell in the range of cardioprotector>nitroglycerin>BOOM. It is seen that the size of THEM when using cardioprotector was significantly lower than with the introduction of the BOOM (p<0,05). Although the introduction of cardioprotector THEM was less than when using nitroglycerin, the differences between groups did not reach the level of statistical significance (p 0,53).

With the introduction of drugs after a period of regional ischemia (in the first minute of reperfusion) significant limitation in comparison with the control was under the influence of cardioprotector and BOOM. Significant differences in limiting THEM with the introduction of nitroglycerin compared with controls were observed (p-value of 0.066). The average values of the size THEY were minimal during the introduction of cardioprotector: 20±1% lower than in the group GEEK (p<0,05).

The results obtained demonstrate the advantages of using cardioprotector, especially at the early stage of reperfusion, to limit THEM in comparison with traditionally used for this purpose drugs.

Example 2. The metabolic state of the heart during reperfusion.

At the end of reperfusion in the hearts of the studied groups were defined levels of high energy phosphates (ATP, ADP, AMP and RKF), creatine (CR) and end products of glycolysis (lactate and pyruvate) to assess the impact of drugs on the metabolism of the myocardium. Based on these data were calculated indicators of energy status postischemic heart:

the energy charge of cardiomyocytes - EZ=(ATP+0,ADF)/Σ;

the degree of phosphorylation of ATP and phosphocreatine - ATP/ADP,

phosphocreatine/creatine (RKF/Kr);

the ratio of lactate/pyruvate, characterizing the degree restore the value of the cytoplasmic peritendinitis [NAD +]/[NADH] [24].

The comparison of obtained data with metabolic state of the heart in the initial state (before the simulation ischemia and reperfusion) are given in table 2.

0,67±0,03ABC
Table 2
The impact of intravenous cardioprotector and Comparators before occlusion of the PNA on the metabolic state of the SP at the end of reperfusion in rats in vivo.
EZ=(ATP+0,ADF)/ΣAHATP/ADPRKF/CuThe lactate/pyruvate
Initial state (n=6)0,94±0,028,03±0,060,65±0,039,8±0,7
Control (n=12)0,60±0,01and1,31±0,02and0,19±0,02andto 98.6±4,6and
Cardioprotector (n=12)0,76±0,02a b1,84±0,04a b0,36±0,03a bto 72.3±4,6a b
GEEK (n=12)1,32±0,03AB0,22±0,03AB87,4±3,1and
Nitroglycerin (n=12)0,71±0,02AB1,44±0,03ABC0,25±0,02ABC78,8±3,9AB
Data are presented as M±M. Control - introduction of saline. Significantly different (p<0,05) from:andinitial state;bcontrol;incardioprotector.

From the data presented in table 2, it is seen that the control energy state WR by the end of reperfusion was significantly reduced compared with the original state of the heart. This is indicated by the lower values of EZ and relations of ATP/ADP and RKF/Cu and more than an order of magnitude increased ratio of lactate/pyruvate. The introduction of cardioprotector or both of the Comparators was significantly improved most indicators of metabolic status SP, bringing them to the values in the initial state. The exception was the ratio of lactate/pyruvate in the BOOM, which did not differ significantly from the values in the control. From the analysis of the indicators that the recovery of the metabolic status of the LA after in the edenia drugs was improved in a series of BOOM< nitroglycerin<cardioprotector and consistent with their ability to limit the size of THEM (table 1). Moreover, under the effect of the introduction of cardioprotector recovery EZ cardiomyocytes was significantly better than in the BOOM, and the relationship of ATP/ADP and RKF/CR was significantly higher than after use of any of the Comparators. The obtained results indicate that the use of cardioprotector to ensure more efficient recovery of aerobic metabolism in LA during reperfusion.

Example 3. Damage to the membrane of cardiomyocytes.

Markers of damage to the sarcolemma postischemic cardiomyocytes served lactate dehydrogenase activity (LDH) and the MB fraction of creatine phosphokinase (MB-ck) in the plasma. Comparison of the increase in the activities of both enzymes were studied at the end of reperfusion in comparison with the values in the initial state are presented in table 3.

Table 3
The impact of intravenous cardioprotector and Comparators before occlusion of the PNA on the activity of LDH and MB-CPK in the blood plasma of rats in vivo.
Initial stateThe end of reperfusion
Control (n=12)71,2±7,7905,9±46,0
Cardioprotector (n=12)72,9±6,9656,3±26,2and
Nitroglycerin (n=12)70,3±5,2750,0±31,6a b
GEEK (n=12)72,9±7,5746,1±35,2a b
MV-ck, IU/l
Control(n=10)430,2±26,01868,0±64,3
Cardioprotector (n=12)428,3±20,11130,1±47,2and
Nitroglycerin (n=10)429,0±34,81323,8±51,7a b
GEEK (n=10)425,2±24,51556,8±60,3a b
Data are presented as M±M for a series of 10 to 12 experiments. Significantly different (p<0,05) from: controland; cardioprotectorb.

In the initial state, the activity of LDH in the plasma was practically the same in all the studied group is Oh (table 3). The development of THEM was accompanied by a significant (12 times) increased activity of LDH in the plasma by the end of reperfusion in the control compared with the original value. The introduction of cardioprotector, as well as comparative drugs before regional ischemia significantly reduced the release of the enzyme into the bloodstream. The minimum increase in the activity of LDH was detected using cardioprotector. It was significantly lower than after the introduction of the BOOM and nitroglycerin, and was about 70% from the value in the control.

In the initial state was no significant differences between groups in activity MB-CPK. By the end of reperfusion activity MB-CPK in the control was increased more than 4 times compared with the original value. A significant reduction of the activity of MV-ck in plasma at the end of reperfusion compared with the levels in control has occurred under the action of all drugs. However, in the case of the introduction of cardioprotector enzyme activity was lower at 15 and 28%higher than in the group of nitroglycerin and BOOM, respectively (p<0,05).

Thus, the use of both markers of damage to cell membranes (LDH and more specific - MB-CPK) clearly indicates greater stability of the sarcolemma postischemic cardiomyocytes SP in the case of cardioprotector proposed composition.

Example 4. Change is hemodynamic in rats in conditions of regional ischemia and reperfusion of the heart.

Action of drugs on the indicators of the functional hemodynamics in rats in vivo was studied when administered before coronary artery occlusion. Drugs and saline were injected, as described in "restricting the size of myocardial infarction". The results are presented in figure 1 and in table 4.

When analyzing the dynamics of blood pressure were identified: 1) lack of vasodilatating response after the introduction of the BOOM as opposed to cardioprotector and nitroglycerin, 2) less sharp drop in Map. in the first phase of activity compared with nitroglycerin, 3) the ability of cardioprotector to prolonged hypotensive effect in combination with the full restoration of BPA. by the end of reperfusion (figure 1).

The analysis of changes in heart rate were found: 1) significantly less decline with the introduction of cardioprotector during occlusion of the coronary artery than in the case of nitroglycerin, 2) a significant decrease in the duration of arrhythmias (on average 3 times) during regional ischemia in comparison with both Comparators (table 4).

Table 4
The effect of intravenous nitroglycerin, BOOM and cardioprotector before occlusion of the PNA on heart rate and duration of drug is making rhythm during ischemia and reperfusion of the heart in anesthetized rats in vivo.
HR, beats/min
Initial state1-I min occlusion of the Palestinian authorityAt 60 min of reperfusionThe duration of the arrhythmia, s
Control224±6228±4229±6445±30
Nitroglycerin222±5183±4and227±7482±42
GEEK230±5225±6224±4518±51
Cardioprotector226±7211±5a b222±5205±13a b
Presents M±M for a series of 10 experiments. Significantly different (p<0,05) from: original status and controland; BOOM and nitroglycerinb.

Thus, a comparison of the actions of drugs comparison with the influence of cardioprotector on dokazateljstvami in rats in vivo evidence on the benefits of using the latest in regional myocardial ischemia and reperfusion al. They are smaller hemodynamic while maintaining prolonged vasodilatation effect and the reduced duration of the arrhythmia. Taken together, the results suggest a promising use of cardioprotector specified composition for reducing ischemic and reperfusion injury of the myocardium.

Variation of the concentrations of the components of cardioprotector.

Were tested two modified composition of cardioprotector containing lower and higher concentration of the main active substances - d-glucose, potassium and pollumassiivi salts of l-aspartic acid, dnci-GLn. The final pH of cardioprotection was straight and was 7.4±0,1 at 22°C (table 5). The effectiveness of these drugs were evaluated in the regional model THEM and reperfusion in rats in vivo to limit the size of myocardial infarction with simultaneous measurement of hemodynamic parameters and activity fractions MV-ck in plasma.

Table 5
The composition of cardioprotection edited content component.
ComponentContent
cardioprotector-1 cardioprotector-2
d-Glucoseto 12.030,0
Potassium chloride0,100,20
Potassium salt of l-aspartic acid0,362,25
Polomania salt of l-aspartic acid0,322,0
Insulin human genetically engineered30,0120,0
Dinitrosyl complex of iron (II) with glutathione0,850,52
Water86,3765,03
Final pH (22°C)7,4±0,17,4±0,1
The content of the components listed in mass %, insulin, IU/l

The data of table 6 show that in the series of experiments with cardioprotection-1 and cardioprotection-2 the values of LA/LV weight (in %) did not differ from those in the control and in the experiments with cardioprotection the optimal composition. Reduced, so is e and the increase in the content of the main active components, increased respect THEM/SP (%) 1.2 and 1.3 times, respectively, compared with the rate when using cardioprotector the optimal composition. In both cases, this led to no significant reduction in the size of infarction compared with control, that is, to the loss of protective actions.

Table 6
The effect of changes in the composition of the components of the drug to limit the size of myocardial infarction in rats in vivo.
The route of administration of medicationThe LA/LV weight, %HIS/SP %
Control (n=12)34,0±3,347,3±2,1
Cardioprotector optimal composition (n=12)before occlusion of the PNA35,5±3,732,4±2,3and
Cardioprotector-1 (n=8)34,6±3,839,8±3,4b
Cardioprotector-2 (n=8)37,0±4,0 42,1±4,1b
Data are presented as M±M. Control - introduction of saline. Significantly different (p<0,05) from:andcontrol;bcardioprotector.

Figure 2 shows the changes in Map. in rats during ischemia and reperfusion of the heart with the introduction of cardioprotector-1 and cardioprotector-2. It is seen that the decrease dnci-Get in cardioprotector-1 2.5-fold compared with cardioprotection optimal composition significantly reduced vasodilatation effect of the drug in the first phase after the introduction and cancelled his prolonged hypotensive effect. In this case the Map. it was restored to its original value at the early stage of reperfusion and was not significantly different from values in control, that is, with the introduction of saline solution. This dynamic Map. pointed to the insufficient allocation of NO dncg and the absence of a binding complex with plasma proteins and erythrocytes [25]. On the contrary, when the introduction of cardioprotector-2 with a 2.5 times increased content dnci-GLT during the first minute was a drastic decrease of BPA. to 59±3% from baseline. By the end of regional ischemia of BPA. remained reduced to 68±4% and did not recover in the future, accounting for 79±4% from baseline at the end of reperfusion. It is obvious that udalenie excess of NO dnci-Get could be the cause lethal damage to cardiomyocytes risk zones [23] and, as a consequence, increase the size of the infarct (table 6).

In series of experiments with the introduction of cardioprotector-1 and cardioprotector-2 was found to decrease the duration of arrhythmias during regional ischemia. The duration of the arrhythmia was not significantly different from values in control and was 498±40 and 668±62 seconds for cardioprotector-1 and cardioprotector-2, respectively, thus exceeding the figure 2.5 and 3.3 times for optimal drug composition (205±13 sec, table 4).

The results of determining the activity of MB-CPK in the blood plasma of rats in vivo with the introduction of cardioprotection with a changed composition of the components shown in table 7. It is seen that the use of cardioprotector-1 and cardioprotector-2 did not reduce the activity of the enzyme at the end of reperfusion compared with the control. In these cases, the activity of this marker of damage to cell membranes was on average 1.6 times higher than in the experiments with the introduction of cardioprotector the optimal composition. These data indicated no reduction breaks proteins in postischemic cardiomyocytes, which agreed well with increasing size of THEM under the influence of cardioprotection with a changed composition components (table 6).

Table 7
The impact of intravenous cardioprotection with modified composition components prior to occlusion of the PNA on the activity of MB-CPK in the blood plasma of rats in vivo
Initial stateThe end of reperfusion
Control (n=10)430,2±26,01868,0±64,3
Cardioprotector optimal composition (n=10)428,3±20,11130,1±47,2and
Cardioprotector-1 (n=8)429,0±34,81704,8±59,0b
Cardioprotector-2 (n=8)425,2±24,51932,8±64,4b
Data are presented as M±M for a series of 8-10 experiments and expressed in IU/L. Significantly different (p<0,05) from: controland: cardioprotectorb.

The obtained results indicate that the variation of components does not increase myocardial protection from ischemic and reperfusion damage. On the contrary, these changes cause the cancellation of the protective action of the drug on all studied parameters. Thus, the best I have is the composition of cardioprotector, specified on page 4. It provides a higher limit of myocardial infarction, the best restoration of the metabolic state of the heart during reperfusion, the most stable membranes postischemic cardiomyocytes and less hemodynamic compared to traditionally used drugs.

Sources of information

Water-soluble composition having the properties of cardioprotector, intravenous, including d-glucose, potassium chloride, insulin human genetically engineered and water, characterized in that it further comprises a potassium salt of 1-aspartic acid, polymagnet salt of 1-aspartic acid and the dinitrosyl iron complex(II) with glutathione in the following ratio of ingredients, wt.%:

d-glucose23,40-28,60
potassium chloride0,14-0,16
potassium salt of 1-aspartic acid0,81-0,99
polomania salt of 1-aspartic acid0,72-0,88
insulin human genetically engineered (IU/l) 54,00-of 66.00
dinitrosyl complex of iron(II) with glutathionefor 0,19 0,23
water for injectionthe rest,

at this pH 7,4±0,1 at 22°C.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of medicine, namely to pharmaceutics in cardiology and can be applied for creation of hypotensive medication from vegetable raw material for treatment of teenagers with essential arterial hypertension. Hypotensive medication for treatment of teenagers with essential arterial hypertension includes dried milled natural raw material: grass of creeping thyme, grass of motherwort shaggy, grass of marsh cudweed, grass of stachys biacalensis fisch, fruits of black chokeberry with the following component ratio, g: creeping thyme, grass 20.0; motherwort shaggy, grass 20.0 marsh cudweed, grass 20.0; stachys biacalensis fisch, grass 20.0; black chokeberry, fruits 30.0. Water infusion is taken in dose 50 ml. 2 times per day in the morning and in the evening, 30 minutes after meal, course of treatment constitutes 3 weeks.

EFFECT: claimed hypotensive medication is simple in application, does not require complex technology of preparing, therefore can be applied in stationary and polyclinic conditions and does not have side effects.

2 cl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to an agent for chronic venous insufficiency. The agent for treating chronic venous insufficiency contains dihydroquercetin and lipoic acid in the ratio 1:1 (weight parts).

EFFECT: agent is effective for chronic venous insufficiency.

1 dwg, 4 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly a drug for glycogen synthesis stimulation in a cardiac muscle. The drug for glycogen synthesis stimulation in the cardiac muscle which represents an alcoholic extract of a porcine and foetal adrenal cortex. A method for glycogen synthesis stimulation in the cardiac muscle by parenteral introduction of the alcoholic extract of the porcine and foetal adrenal cortex in a certain dose.

EFFECT: agent effectively stimulates glycogen in the cardiac muscle with no side effects.

2 cl, 2 dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, specifically cardiology and concerns treating ischemic heart disease (IHD). For this purpose, the standard integrated drug treatment involving statin is added with administration of the biologically active additive MARISTIM 4.5 mg/day.

EFFECT: method provides higher clinical effectiveness of statins in the given group of patients due to elimination of their inhibitory effect on enzymes of a mitochondrial respiratory chain.

2 cl, 1 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a method of treating hypertension, congestive cardiac failure, stenocardia, myocardial infarction, atherosclerosis, stroke. A method of treating involves introduction to a patient requiring such treatment of the solid oral dosage form containing a therapeutically effective amount of aliskiren or its pharmaceutically acceptable salt in which the active ingredient makes more than 46 wt % of total weight of the oral dosage form. The oral dosage form is presented in the form of a tablet or a film-coated tablet and produced by methods other than wet granulation with excipients by means of water and/or a water solution of a binding agent.

EFFECT: realisation of the specified purpose.

12 cl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a method of treating hypertension, congestive cardiac failure, stenocardia, myocardial infarction, atherosclerosis, stroke. A method of treating involves introduction to a patient requiring such treatment of the solid oral dosage form containing a therapeutically effective amount of aliskiren or its pharmaceutically acceptable salt in which the active ingredient makes more than 46 wt % of total weight of the oral dosage form. The oral dosage form is presented in the form of a tablet or a film-coated tablet and produced by methods other than wet granulation with excipients by means of water and/or a water solution of a binding agent.

EFFECT: realisation of the specified purpose.

12 cl, 2 ex

FIELD: medicine.

SUBSTANCE: for the purpose of hypoestrogen-induced endothelial dysfunction correction in white female Wistar rats, hypoestrogen-induced endothelial dysfunction is simulated by bilateral ovariectomy. The dysfunction correction is ensured by introduction of mixed solutions of homoeopathic dilutions of polyclonal rabbit C12, C30, C200 antibodies to human endothelial nitrogen oxide synthase for 6 weeks following the ovariectomy. The specified mixture is added to drinking bowls once a day.

EFFECT: method provides effective correction with reducing an endothelial dysfunction coefficient in females to the level observed in intact animals ensured by the effect on various developmental mechanisms of this dysfunction in hypooestrogenic conditions.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: for the purpose of hypoestrogen-induced endothelial dysfunction correction in white female Wistar rats, hypoestrogen-induced endothelial dysfunction is simulated by bilateral ovariectomy. The dysfunction correction is ensured by daily intragastric introduction of enalapril 0.5 mg/kg for 6 weeks following the ovariectomy. The mixed solutions of homoeopathic dilutions of polyclonal rabbit C12, C30, C200 antibodies to human endothelial nitrogen oxide synthase are also introduced. The specified mixture is added to drinking bowls once a day.

EFFECT: method provides effective correction with reducing an endothelial dysfunction coefficient in females to the level observed in intact animals ensured by the effect on various developmental mechanisms of this dysfunction in hypooestrogenic conditions.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: endothelial dysfunction is ensured by intragastric introduction to a laboratory animal of the NO-synthase inhibitor - N-nitro-L-arginine of methyl ester (L-NAME) 25 mg/kg daily for 7 days. The endothelial dysfunction correction is ensured by intragastric introduction of atorvastatin 2.2 mg/kg and intragastric introduction of resveratrol 2 mg/kg once a day.

EFFECT: effective action of the combination of two specified preparations on vascular endothelium function.

1 ex, 2 tbl

FIELD: medicine.

SUBSTANCE: endothelial dysfunction is ensured by intragastric introduction to a laboratory animal of the NO-synthase inhibitor - N-nitro-L-arginine of methyl ester (L-NAME) 25 mg/kg daily for 7 days. The endothelial dysfunction correction is ensured by intragastric introduction of atorvastatin 2.2 mg/kg and intragastric introduction of resveratrol 2 mg/kg once a day.

EFFECT: effective action of the combination of two specified preparations on vascular endothelium function.

1 ex, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, particularly, to a conjugate of an insulinotropic peptide of prolonged action in vivo and stability, and containing an insulinotropic peptide selected from a group consisting from exendin-3 and exendin-4 and their derivatives, a nonpeptide polymer where one of the ends of a molecule of the nonpeptide polymer is connected with an amino acid residue different from a N-terminal residue of the insulinotropic peptide.

EFFECT: invention provides preparation of the conjugate of the insulinotropic peptide exhibiting activity in vivo which remains at a relatively high level, and substantially increased blood half-life, availability for development of the long-acting formulations based on various peptide drugs.

29 cl, 1 tbl, 18 dwg, 15 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to pharmacy and medicine. Pharmaceutical composition includes mixture of: (a) active macromolecular substance; (b) aromatic alcohol, absoption amplifier, selected from propylgallate, butylated hydroxytotuol (BHT), butylated hydroxyanisole (BHA) and their analogues and derivatives, or their mixtures; and (c) biguanide or its pharmaceutically acceptable salt, capable of increasing solubility of aromatic alcohol, absorption amplifier, in water medium, where aromatic alcohol, absorption amplifier, is present in amount (by weight) greater or equal to amount of active substance. Application in pharmaceutical composition of aromatic alcohol, selected from propylgallate, BHT, BHA and their analogues and derivatives, together with biguanide or its pharmaceutically acceptable salt, capable of increasing solubility of aromatic alcohol in water medium, as amplifier of macromolecule absorption through intestine wall. Method of increasing absorption of active macromolecular substance and method of patient treatment include introduction of claimed composition to patient. Application of active macromolecular substance, selected from insulin, C-peptide, GLP-1 or their mixture; aromatic alcohol, absorption amplifier, selected from propylgallate, butylated hydroxytotuol (BHT), butylated hydroxyanisole (BHA) and their analogues and derivatives, or their mixtures; and biguanide or its pharmaceutically acceptable salt, capable of increasing solubility of aromatic alcohol, absorption amplifier, in water medium, in manufacturing medications for treatment of diabetes, osteoporosis, obesity, cancer, osteoarthritis.

EFFECT: group of inventions ensures facilitation of passage of peptides, proteins and other macromolecular substances through intestinal wall for improvements of their bioavailability.

27 cl, 3 tbl, 11 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new polysaccharide protein derivatives, particularly insulin where polysaccharide is polysialic acid containing 20 to 125 units of sialic acid and where at least 85 % of insulin derivatives are modified on a N-tail of insulin B-chain only, and also to methods of producing it.

EFFECT: invention provides higher stability, prolonged biological half-life, lower insulin immunogenicity, and can find application in pharmaceutical compositions for diabetes treatment.

3 tbl, 15 dwg

FIELD: medicine.

SUBSTANCE: claimed is medication for treatment of blood loss, consisting of complex representing combination of insulin 14 U, glucose solution 10% - 400 ml, sodium adenosine triphosphate 1% - 1 ml, panangin 10 ml, strophanthin acetate 0.05% - 0.25 ml, ascorbic acid 5% - 2 ml, riboxin 10ml, analgin 25% - 1 ml, polyglucinum 6% -400 ml.

EFFECT: medication in express-mode simultaneously produces rehabilitating action on all factors of blood loss etiopathogenesis, including elimination of acidosis, stress and pain syndrome.

1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology and medicine, and represents a method of producing an oral preparation of proinsulin C-peptide of activated polyethylene glycol differing by the fact that polyethylene glycol is activated by ionising radiation in an acidic medium with calcium and/or zinc cations added to final concentration 5-10 mM, and then mixed with proinsulin C-peptide, polyethylene glycol is characterised by molar mass 0.4-40 kDa and concentration 1.0-50.0%, in relation of proinsulin C-peptide: polyethylene glycol equal to 1:(1-500) to final concentration of proinsulin C-peptide in mixture 1-10 mg/ml.

EFFECT: invention provides higher therapeutic activity in oral administration with respect to diabetic complications.

4 cl, 3 ex,1 tbl, 2 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to psychiatry, and deals with treatment of schizophrenia, resistant to pharmacotherapy. For this purpose insulin coma therapy is carried out, 20 mg of propranolol being introduced to patient sublingually immediately after each subcutaneous injection during the whole course of therapy.

EFFECT: method ensures increase of efficiency and reduction of treatment terms due to potentiating insulin action by propanolol at the background of reduction of side effects frequency, which is conditioned by elaborated mode of introduction of medications.

1 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, biotechnology and medicine, particularly to methods of producer cell destruction, and can be used in insulin substances production. A commercial production process of human recombinant insulin involves the bacterial cell destruction; the bacterial cell destruction and following filtration are performed in a nanohomogeniser. The nanohomogeniser 'Microfluidiser' is used as said nanohomogeniser. The bacterial cell destruction is performed at pressure 2070 bar and flow rate 330-450 ml/min.

EFFECT: use of the method provides higher efficiency and reduced length of the technological commercial insulin production process; the recombinant protein granule yield is 98%, the one-stage cell destruction is enabled, the granules of the same nanosize, particularly 60-80 nm are produced.

1 dwg, 1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine and may be used to heal wound epithelium of various genesis. For this purpose an agent is introduced into wound, causing modulation of expression and/or activity of one isoform PKC and therapeutically effective amount of one additional facility selected from group that consists of insulin, growth factor, adipokin, PKC RACK and GW9662. According pharmaceutical compositions are proposed for introduction in wound, containing, apart from therapeutically effective amounts of necessary substances, a pharmaceutically acceptable carrier.

EFFECT: accelerated processes of epithelium wound healing by explicit synergetic action of specified substances.

60 cl, 43 dwg, 10 tbl, 27 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: method of obtaining dry diketopiperaxine powder includes obtaining diketopiperazine solution, stage of diketopiperazine-containing particles formation and removal of solvent by drying by means of dispersion. Method can include additional stage of loading diketopiperazine particles with active agent, which represents peptide or protein, preferably, insulin. Diketopiperazine preferably is fumaryldiketopiperazine. Diketopiperazine particles, obtained by invention method, possess improved aerodynamic characteristics, active agent, in particular insulin, is more stable and is delivered in more efficient way in comparison with composition of diketopiperazine-insulin, dried by means of sublimation.

EFFECT: dry powders are used as compositions of pharmaceutical preparations for delivery in pulmonary way.

16 cl, 10 dwg, 5 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: invention refers to pharmacology and medicine, particularly to endocrinology. A method for making proinsulin preparation consists that in 1-50 % aqueous polyethylene glycol with molar mass 0.4-40 kDa pre-exposed to ionising radiation in amount 1.0-5.0 Mrad in acid medium at pH 2.5 - 4.5 with calcium and/or zinc cations added to final concentration of 5-10 mm, proinsulin is introduced to final concentration of 1-10 mg/ml in the ratio of polyethylene glycol to proinsulin equal (1-500)/1. The mixture is stirred to prepare a homogeneous transparent or slightly opalescent solution.

EFFECT: use of the method allows simplifying process for making proinsulin preparation and ensuring possibility of its application as a hypoglycaemic agent and maintenance of activity at pH 2-3 and room temperature.

4 cl, 1 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, and concerns a method for making a hemodialysis solution used in extracorporal blood purification for treating patients with acute and chronic renal insufficiency. The hemodialysis solution contains the following components, mmol/l: Sodium chloride NaCl 137.3-140.3; Calcium chloride CaCl2·2H2O 1.2-1.75; Magnesium chloride MgCl2·6H2O 0.5-1.0; Potassium chloride KCl 0.3-4.0; Glucose, g/l 1.0-5.0; Hydrochloric acid 0.1-2.9; Amber acid C4H6O4 0.1-2.9; Sodium bicarbonate 32.0.

EFFECT: invention aims at producing a physiologic composition of an acid component with no acetic acid added, providing minimum complications in the form of acidosis, and haemodynamic stability in patients of a risk group and a cardiac intensive profile.

4 ex

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