RU2549448C1 - Balanced infusion solution - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents a balanced infusion solution containing sodium, potassium and magnesium chlorides, a solvent and sodium L-arginine succinate of formula: Na⁺[NH=C(NH₂)NH(CH₂)₃CH(NH₂)COOH]⁺[OOC(CH₂)₂COO]^2-. The ingredients in the solution are found in certain proportions, wt %.

EFFECT: invention provides enhanced detoxification activity, low toxicity and wide range of clinical applications.

11 tbl, 6 ex

Description

The field of technology.

The invention relates to medicine and pharmacology, in particular to saline infusion solutions with a detoxifying effect, and can be used in the treatment of diseases and conditions associated with intoxication of the body of varying severity.

The prior art.

In medical practice, salt infusion solutions based on succinic acid salts are widely used.

For example, a polyionic solution based on sodium succinate - “Reogemin” (Registration Certificate of the Republic of Belarus RU 12/12/2047 of 12/04/12), used as an antihypoxic and detoxification agent for acute endogenous and exogenous intoxications of various etiologies and for the correction of water- salt balance in the body containing the following components: sodium succinate hexahydrate 7.5 g / l, sodium chloride 4.0 g / l, potassium dihydrogen phosphate 1.4 g / l, magnesium chloride hexahydrate 0.02 g / l, glucose 50 g / l and has the following ionic composition: sodium 161 mmol / l, potassium 10.2 mmol / l, magnesium 1.1 mmol / l, chlorides 70.5 mmol / l, phosphates (in terms of phosphorus) 10.0 mmol / l (Instructions for medical use , order of the Ministry of Health of the Republic of Belarus No. 1515 of 12/26/12)

It must be emphasized that this solution is not balanced for the main vital ions and contains an excess of sodium (by 10%), an excess of potassium ions (by 250% of the norm), and a lack of chlorine ions (by 30%), which creates correction problems water-salt metabolism in patients.

In addition, it is known that the combination of glucose solutions with low molecular weight dicarboxylic acids (in this case with succinic acid) poses a risk of impurities (furfurols) hazardous to the health of patients during production and storage of the drug (Encyclopedia of Pharmaceutical technology. Third Edition, 2006).

Also known is the drug "Succinasol" containing sodium chloride - 6.2 g; potassium chloride - 0.3 g; calcium chloride - 0.082 g; magnesium chloride - 0.1 g; succinic acid - 2.0 g; sodium bicarbonate - 3.0 g; water for injection - up to 1 l) and has the following ionic composition: sodium 142 mmol / l, potassium 4.0 mmol / l, magnesium 1.1 mmol / l, chlorides 114 mmol / l, proposed as a means for treating critical conditions and, also containing in its composition as an active component succinic acid. “Succinasol” improves blood microcirculation, activates energy metabolism, restores hemodynamic parameters and cardiac muscle function in various pathological conditions (patent UZ 4106 В from 04/21/1994, RU 10/140/3 from 05/11/10, Levin GS, Shevchenko LI, Gubaev SA. Comparative evaluation of the effects of crystalloid solutions containing sodium lactate and sodium succinate on the hemodynamic indicators in different types of hemorrhage, Gematol Transfuziol. 1991 Feb; 36 (2): 25-8., PMTO: 2055408).

Succinate solutions are known in combination with a complex of vitamins and metabolic components. Thus, the drug “Cytoflavin” is known (Registration number of the Russian Federation PN 00313/1 dated October 21, 2004) containing succinic acid in the form of sodium meglumine succinate, in combination with inosine, nicotinamide and riboflavin, which has a cytoprotective effect (patent EA 001099).

In clinical practice, the Reamberin polyionic infusion solution (Registration number of the Russian Federation R No. 99/363/2 10/08/1999) is used, which contains succinic acid in the form of sodium meglumine succinate and a balanced set of macronutrients of potassium, magnesium, sodium salts and used as a detoxifying drug (patent EA 000879) and Remaxol infusion solution (Registration number of the Russian Federation LSR-009341/09 of 11/19/2009) containing succinic acid in the form of sodium meglumine succinate in combination with inosine, nicotinamide and methionine having hepatoprotective action (patent EA 007865).

The closest analogue of the claimed invention in pharmacological action and composition is the salt infusion solution "Reamberin" (patent EA 000879), selected as a prototype. The drug is balanced in ionic composition and has detoxifying properties due to the normalization of water-electrolyte balance, antihypoxic and antioxidant effects.

However, despite the presence of a high therapeutic effect, the known infusion solution has a key pharmacological drawback, due to the peculiarity of the active component of meglumine sodium succinate, the cation of which (methylglucammonium) practically does not metabolize in the human body and, when the anion is used, monosubstituted sodium succinate is able to accumulate in the body and lead to significant alkalization of blood (alkalosis) and urine, as noted in the instructions for medical use of the prototype (Register National number of the Russian Federation R No. 99/363/2 10/08/1999). This condition is difficult to correct and is especially dangerous in patients with reduced blood buffer capacity and renal failure in case of serious critical conditions such as polytrauma, sepsis, frostbite of organs and tissues, liver failure, and other conditions associated with activation of catabolic processes in the body.

In this regard, the high sodium content of meglumine succinate in the prototype limits the maximum daily volume of administration of this drug to 800 ml, which is insufficient for infusion therapy of a number of critical conditions (acute blood loss, sepsis, polytrauma, hepatic encephalopathy) requiring the introduction of large volumes (up to 10 liters a day).

In addition, alkalization of urine, characteristic of the use of the prototype and due to the excretion of the organic base of methylglucammonium through the kidneys, can be dangerous for patients with nephrolithiasis. This is due to the possibility of solubilization of calcium urate and oxalate under the influence of methylglucammonium, which can initiate the migration of kidney stones and blockage of the ureters.

All of the above disadvantages of the known infusion solution limit its clinical use in high doses and for certain groups of patients, which significantly narrows the scope of its application.

A brief description of the invention.

The objective of the invention is to create a new balanced infusion solution with increased detoxifying activity, low toxicity and providing a wide area of clinical application.

The problem is solved in that a balanced infusion solution containing sodium, potassium and magnesium chlorides, a solvent and a biologically active component, according to the invention, contains sodium L-arginine succinate of the formula as a biologically active component:

in the following ratio of components, wt.%;

Sodium chloride 0.540-0.600 Magnesium Chloride Hexahydrate 0.015-0.030 Potassium chloride 0,025-0,040 Sodium L-Arginine Succinate 1,400-1,700 Water for injections Rest

The inventive infusion solution contains sodium chloride, magnesium chloride hexahydrate and potassium chloride in a certain proportion, providing a physiologically acceptable content of inorganic ions of sodium, potassium, magnesium and chlorine (Magu Lee, Basic skills in interpreting laboratory data., 4 ^th ed.).

As a biologically active component of the inventive balanced infusion solution contains a new chemical compound of sodium L-arginine succinate of the formula:

Physico-chemical properties of sodium L-arginine succinate are shown in table 1:

Good solubility in water and a physiologically acceptable pH value of aqueous solutions (7.0-7.2) of sodium L-arginine succinate, makes it suitable for the preparation of parenteral drugs.

It is known that salts of succinic acid in a composition with a conditionally essential amino acid L-arginine are used as components of drugs with stabilizing and buffering properties. For example, a tissue factor pathway inhibitor and L-arginine succinate composition is known (patent EP 1602667), for preserving antibody solutions (application US 20100239567, JP 2009-108040, application US 20120121581) and as a detergent and cosmetic product (application JP 2001 -081025).

Complex compositions are described, which include L-arginine and succinic acid for the treatment and prevention of hepatic encephalopathies in the form of effervescent tablets used as biological food additives (application RU 2011109884/15).

The scientific and technical literature describes the cardioprotective effect of mono- and di-argininosuccinates - various combinations of succinic acid with L-arginine.

Thus, the effect of L-arginine monosuccinate as a potential cardioprotective drug was established (B.V. Dubovik, A.A. Zhdanov, D.I. Romanovsky, Comparative cardioprotective efficacy of arginine hydrochloride and arginine succinate in ischemic reperfusion injury of rat myocardium, 2005, Medical Journal of the Belarusian State Medical University No. 4, S. 51-53).

On the basis of diargininosuccinate, the drug “Cardinosine” was registered (TU BY 100185198.112-2009, registration number 027761 dated 12/09/2009) containing the active substance in the form of a 5% aqueous solution, used as an agent for the treatment of acute myocardial infarction.

In a number of inventions, combined drugs have been proposed for the treatment of cardiovascular diseases based on a combination of arginine succinate and arginine asparaginate in the form of a solution for oral parenteral use (patents UA No. 36998, 37388, 39095, 90368).

The available scientific and technical literature does not describe infusion solutions containing, as the active component of sodium, L-arginine, succinate of the formula (1).

The present invention is the first to show that a new chemical compound of formula (1) can be used as a biologically active component of a detoxifying infusion solution.

In particular, the authors found that the introduction of sodium L-arginine succinate in the composition of the inventive infusion solution increases its detoxifying properties and reduces toxicity compared to the prototype.

As a result of experiments on the selection of a balanced composition of the proposed infusion solution, it was found that solutions obtained in the range of sodium L-arginine succinate from 14 to 17 g / l have a theoretical (calculated) osmolarity of 350 to 360 mmol / l and practical (physiological ) osmolality 315-324 mmol / kg, respectively. It is known that the actual osmolality of blood plasma is 286 ± 5 mmol / kg. However, the osmolality in the range of 315-324 mmol / kg for the inventive infusion solution is optimal, since the pharmacokinetics data showed that in vivo sodium L-arginine succinate quickly penetrates the cells and metabolizes to osmotically inactive carbon dioxide and water, which reduces it osmolality to physiological values.

These characteristics make it possible to balance the composition of the claimed infusion solution and introduce it without significant volume restrictions, which expands the scope of clinical use by eliminating a number of contraindications associated with alkalization of blood and urine.

DETAILED DESCRIPTION OF THE INVENTION

In the inventive infusion solution, the ratio of the components ensuring the balance of the composition was selected empirically.

So, the introduction of potassium and magnesium cations to the composition of the claimed infusion solution is proposed by adding potassium chloride in concentrations of 0.025-0.040 g / l and magnesium chloride hexahydrate in concentrations of 0.015-0.030 g / l, providing a physiologically acceptable content of potassium ions 3.4-5, 3 mmol / L and magnesium 0.7-1.5 mmol / L, respectively.

Introduction to the composition of the inventive infusion solution of sodium ions is proposed by adding sodium chloride. For example, the addition of sodium chloride at a concentration of 8.6 g / l makes it possible to obtain a physiologically acceptable content of sodium cations - 147 mmol / l. In this case, the content of chlorine anions formed during the dissociation of sodium chloride, potassium chloride and magnesium chloride hexahydrate will be about 153 mmol / L, which will significantly exceed the physiological norm (Mary Lee, Basic skills in interpreting laboratory data., 4 ^th ed.).

The present invention proposes to bring the chloride content to a physiological norm (97-110 mmol / L) by replacing part of the inventive infusion solution of a portion of sodium chloride (NaCl) with a new biologically active substance. At the same time, its content in the preparation in concentrations from 14 to 17 g / l provides the physiological content of chlorides without violating the content of sodium cations.

The invention is as follows.

Example. At the first stage, a new biologically active component is obtained - sodium L-arginine succinate of the formula (1).

To obtain sodium L-arginine succinate, 52.80 kg of succinic acid are taken (CAS Number 110-15-6); 77.89 kg of L-arginine (Eur. Ph.); 17.88 kg of sodium hydroxide (Eur. Ph.) And dissolved in 500 l of water. In this case, an aqueous 28% solution of the new sodium substance L-arginine succinate is formed according to the chemical reaction:

The resulting solution was spray-dried and 134.9 kg of sodium L-arginine succinate was isolated (96% yield) in the form of a white amorphous powder, the physicochemical properties of which corresponded to the data given in table 1. Subsequently, the sodium powder of L-arginine succinate packaged in a sealed primary packaging with high barrier properties with respect to water vapor and oxygen, and then into the secondary packaging.

At the second stage receive the claimed infusion solution.

To do this, take a powder of sodium L-arginine succinate in an amount of 14.052 kg and dissolve it in 500 l of water for injection, add 6.0 kg of sodium chloride (Eur. Ph.), 0.244 kg of magnesium chloride and 0.3 kg of potassium chloride to the solution (Eur. Ph.) And adjust the volume of the solution to 1000 L with water for injection (Eur. Ph.). Then the resulting solution is filtered, poured into a glass or plastic package, hermetically closed and sterilized using the standard mode (kept at a temperature of 121 ° C for at least 15 minutes).

The infusion solution made according to the example has the following composition:

A study of the safety and biological activity of the inventive balanced infusion solution obtained in the preparation example was carried out in comparison with the prototype drug Reamberin, a solution for infusion of 1.5%, in a series of experimental studies on animals (experiments 1-6).

Experience 1. The study of acute toxicity of the inventive infusion solution

The experiments were performed on white outbred rats of both sexes. To determine indicators of acute toxicity, the test solution was administered intravenously in accordance with the planned method for the use in clinical practice of rats of both sexes in increasing doses according to Litchfield-Wilcoxon and in the maximum volumes possible for acute administration. Control animals were injected with a 0.9% sodium chloride solution in a volume of 20 ml / kg (ICH M 3 (R2) Guidance of Non-Clinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals, Current Step 4, version dated 11 June 2009).

When studying the acute toxicity of the claimed infusion solution, it was revealed that with the introduction of the drug at the maximum possible dose of 8175 mg / kg in terms of the active substance, mortality was not observed in the group of animals. While the toxicity of the solution of the prototype was LD ₅₀ = 5600 mg / kg body weight of the animal in terms of the active substance.

Thus, in the conditions of acute experience, the claimed infusion solution is less toxic than the prototype, does not have mutagenic, teratogenic, embryotoxic, allergenic and immunotoxic effects.

Experience 2. The study of the effect on mortality of the proposed infusion solution on a model of acute endotoxemia induced by lipopolysaccharide from Salmonella enterica

The experiment was carried out on 200 standard Wistar rats aged 17-18 weeks and weighing 180-210 g. All animals received standard food and water.

Lethal endotoxemia was modeled by the administration of a bacterial endotoxin - Salmonella enterica lipopolysaccharide serotype enteritidis (SIGMA-ALDRICH.). Endotoxin was administered intravenously (in the tail vein) once in 0.2 ml of water for injection. Doses of endotoxin causing a pyrogenic reaction and 50 and 100% mortality of animals from toxic shock, which were 5 μg / kg and 25 μg / kg of endotoxin, respectively, were experimentally selected. The pyrogenic reaction was recorded by measuring rectal temperature (Filkins JP, Di Luzio NR, Endotoxin induced hypothermia and tolerance in the rat. Proc. Soc. Exp. Biol. Med. 1968 Dec, 129 (3): 724-6; Utili R, Abernathy CO, Zimmerman HJ. Endotoxin effects on the liver. Life Sci. 1977 Feb 15, 20 (4): 553-68; Banet M., Fever and survival in the rat. The effect of enhancing fever, Pflugers Arch. 1979 Jul, 381 (1): 35-8.).

For the study, eight groups of experimental animals were formed, 25 rats each: 2 control groups without treatment, 2 experimental groups of animals that were injected with a 0.9% sodium chloride solution at a dose of 20 ml / kg, 2 experimental groups of animals that were injected with a prototype at a dose of 20 ml / kg, 2 experimental groups of animals that were injected with a new infusion solution at a dose of 20 ml / kg. The studied drugs were administered intravenously once 2 hours after the administration of lipopolysaccharide. Evaluation of the effectiveness of the drugs was evaluated by the mortality rate in the groups within seven days after the administration of lipopolysaccharide (table. 2).

In the animals of the control groups, during the first seven days after the administration of lipopolysaccharide at doses of 5 and 25 mg / kg, toxic shock developed - hyperthermia, decreased motor activity, decreased muscle tone, refusal to eat, diarrhea and death.

Administration of endotoxin at a dose of 5 mg / kg resulted in 60% mortality.

Conducting standard therapy of the formed toxic shock with 0.9% sodium chloride solution reduced animal mortality by 1.5 times (from 60% to 40%). The use of the prototype reduced mortality compared with the control group of animals, by 1.88 times (from 60% to 32%), and compared with the group treated with sodium chloride by 1.25 times.

When using the inventive solution, there was a decrease in mortality during the seven days of the experiment compared with the control 2.5 times, compared with sodium chloride therapy - 1.67 times, and compared with the prototype - 1.33 times (from 32% up to 24%).

An increase in a single dose of endotoxin administered to laboratory animals to 25 mcg / kg led to an increase in toxic shock and 100% mortality within seven days of observation.

Conducting standard therapy with a 0.9% sodium chloride solution at a dose of 20 ml / kg reduced the mortality of animals in the group by 1.39 times (from 100% to 72%).

The use of the prototype reduced mortality compared to the control group by 1.47 times, and compared with the group receiving sodium chloride by 1.06 times.

The use of the proposed solution shows a more significant reduction in mortality during the seven days of the experiment compared with the control by 1.67 times, compared with sodium chloride therapy - by 1.2 times, and compared with the prototype 1.13 times (from 68% up to 60%).

Thus, testing the inventive infusion solution in a toxic shock model showed its high detoxifying activity at various doses of Salmonella enterica endotoxin.

Experience 3. The study of the effectiveness of the proposed infusion solution on a model of acute toxic hepatitis

Studies of the proposed solution were carried out in comparison with the prototype in rats of males of the breed "Wistar" with a body weight of 180-200 g. The experiment was conducted on 90 animals of which 6 groups were formed: intact animals (n = 15); control group (n = 15) animals were injected with 0.9% sodium chloride solution at a dose of 20 ml / kg; experimental groups No. 1 (n = 15) and No. 2 (n = 15) animals, which were introduced the prototype in doses of 10 ml / kg and 20 ml / kg, respectively;

experimental groups No. 3 (n = 15) and No. 4 (n = 15) animals that were injected with the inventive infusion solution in doses of 10 ml / kg and 20 ml / kg, respectively.

To form acute toxic hepatitis, the animals of the control and experimental groups were once intraperitoneally injected with phenylhydrazine sulfate at a dose of 50 mg / kg, and then after 24 hours a 50% oil solution of carbon tetrachloride at a dose of 3 ml / kg of body weight was administered intragastrically. After 2 hours in the experimental groups, infusion therapy with the studied solutions was started. The therapy was carried out for 2 days sequentially. At the end of the treatment, intoxication markers were evaluated: the activity of the enzymes alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, alkaline phosphatase, bilirubin level and hemostasis indicators: prothrombin time, activated partial platelet thrombotic time, thrombin time (Heimberg Мotchoterbox tetron, Watkins ML the direct action of CCl ₄ on the liver Pharmacol. Exp. Ther. 1964 Jul 145: 92-101; Plaa GL, Larson RE, CCl ₄ induced liver damage. Current concept, regarding mechanism of action. Arch Environ. Health., 1964 Oct. (9), 536-43; Bruckner JV, Mac Kenzie WF, Muralidhara S, Luthra R, Kyle GM, Acosta D. Oral toxicity of carbon tetrachloride: acute, subacute, and subchr onic studies in rats. Fundam. Appl. Toxicol. 1986 Jan. 6 (1): 16-34).

In the control group, signs of the development of toxic hepatitis were noted: cytolysis syndrome, impaired liver detoxifying function with an increase in the activity of alkaline phosphatase enzymes, aspartate aminotransferase, bilirubin level, “failure” of alanine aminotransferase activity and a tendency to increase lactate dehydrogenase activity (p = 0.222 compared to the group of intact animals) ( table 3).

Prototype infusion therapy helped to limit the cytolysis syndrome by its effect on the activity of alanine aminotransferase and aspartate aminotransferase, but did not affect the activity of the alkaline phosphatase cholestasis marker and bilirubin level. The action of the proposed solution, in contrast to the prototype, was characterized by a significant decrease in the level of bilirubin (p = 0.032) and alkaline phosphatase activity (p> 0.05 compared with the group of intact animals). However, the identified changes were multidirectional (Table 4).

Positive changes during therapy with the prototype and to a greater extent the claimed solution were expressed in a decrease in pathologically increased activated partial thromboplastin time (internal coagulation pathway) and thrombin time. At the same time, against the background of therapy with the claimed solution, a reliable normalizing effect was revealed in relation to the activated partial thromboplastin time and thrombin time (p = 0.009 and 0.05) compared with those of the prototype in a dose equivalent in volume.

Analyzing the obtained data, we can conclude that the claimed infusion solution has a significantly more pronounced hepatoprotective effect in the acute toxic hepatitis model than the prototype.

Experience 4. The study of the effectiveness of the proposed infusion solution on the model of poisoning with ammonium carbonate

The experiments were performed on 150 outbred white male rats weighing 170-180 g at the age of 13-14 weeks. A total of 5 experimental groups were formed: intact animals (n = 30); control group - animals with intoxication without treatment (n = 30); experimental group 1 - animals with intoxication and the introduction of the inventive infusion solution (n = 30); experimental group 2 - animals with intoxication and the introduction of physiological saline (n = 30); experimental group 3 - animals with intoxication and the introduction of the prototype (n = 30).

Ammonium carbonate intoxication was created by introducing its aqueous 10% solution intragastrically through an atraumatic probe at a dose of 250 mg / kg (LD ₅₀ ). The clinical picture with the introduction of the toxicant was accompanied by initial motor excitement, vocalization, followed by a pronounced general toxic effect - a significant decrease in activity, muscle tone, reaction to pain irritation against the background of ataxia and the development of a soporous state. Some animals showed lateral position and cramps. The acid-base balance of animal blood (Table 5) showed the nature of the seizure. The acid-base balance of animal blood (Table 5) showed a characteristic picture of the development of metabolic alkalosis under the action of ammonia released from ammonium carbonate (Karr NW, Hendriks EL. The toxicity of intravenous ammonium compounds. Am. J. Med. Sci. 1949 Sep. 218 (3): 302-7; Ting YC. The toxicity of ammonia. Science. 1950 Jul 21, 112 (2899): 91-2; Warren KS, The differential toxicity of ammonium salts. J. Clin. Invest. 1958 Apr 37 (4): 497-501; Handford SW, An experimental study of ammonium intoxication. Gastroenterology. 1959 Jun; 36 (6): 770-9.).

Compared solutions began to be administered intravenously 1 hour after the development of severe symptoms of intoxication into the tail vein at a dose of 20 ml / kg for 3 days. The effectiveness of treatment was judged by the clinical picture, mortality, indicators of oxygen budget and acid-base balance: NSO ₃ - plasma bicarbonate; pCO ₂ - partial pressure of carbon dioxide; pO ₂ is the partial pressure of oxygen; BB - excess blood bases; BE - deficiency of plasma buffer bases; BE ecf - excess of extracellular fluid bases. In addition, kidney function was assessed by the weight of the kidneys, daily urine output, protein and chloride in the urine, density and pH of the urine.

4 hours after the administration of the studied solutions, 10 animals from each group were decapitated, mixed blood was taken and the parameters of the oxygen budget and acid-base balance were measured (Table 5)

The use of the inventive solution allowed to reduce the excess of buffer bases of blood plasma, to normalize the pH level, pCO ₂ in contrast to the prototype, which, on the contrary, contributed to an increase in the pH level. Moreover, the inventive solution as a whole contributed to a more effective reduction in the manifestations of alkalosis and hypoxia.

By the third day of observation, mortality in the control group was 15 out of 20 animals (75%). Surviving animals gained weight worse, were less active compared with animals of the experimental groups.

The use of physiological saline (Experimental group 2) reduced mortality to 60% (12 animals died 20), and the prototype (Experimental group 3) - up to 20% (4 animals out of 20 died). Treatment with the claimed infusion solution (Experimental group 1) led to the absence of mortality.

The objective condition of the animals was better when using the inventive solution and prototype (table. 6).

In animals of the control group, there was a significant decrease in daily urine output and an increase in the amount of protein and urine pH. The use of the inventive solution, prototype and sodium chloride had a diuretic effect: increased daily diuresis and chloride content in the urine. However, the diuretic effect of the inventive solution was more pronounced than that of the prototype and physiological saline.

The results indicate a high efficiency of the proposed infusion solution in comparison with the prototype in the treatment of conditions accompanied by alkalosis. So, the claimed infusion solution contributes to the normalization of metabolic disorders in the body caused by alkalosis, reduces the severity of manifestations of hypoxia, which positively affects the function of the kidneys and the state of the body as a whole.

Experience 5. The effectiveness of the proposed infusion solution on the model of acute ethanol poisoning

The experiments were performed on 105 male rats. For each experimental model, 7 groups of animals were formed (n = 15).

For the formation of acute poisoning, animals were once injected intraperitoneally with 40 vol. % ethanol solution at a dose of 8 ml / kg body weight. After 2 hours, all experimental animals started the infusion therapy with the studied solutions. The therapy was carried out for 2 days in a row. The solutions were administered three times a day with an interval of 2 hours due to the limitation of the maximum single volume for administration to rat veins. A new infusion solution was administered in doses: 10 ml / kg, 20 ml / kg, 30 ml / kg, 40 ml / kg, and the prototype in a dose effective according to published data - 20 ml / kg per day (Di Luzio NR, A mechanism of the acute ethanol-induced fatty liver and the modification of liver injury by antioxidants. Am. J. Pharm. Sci. Support Public Health, 1966 Jan; 15: 50-63; Acute ethanol intoxication and liver lipid metabolism. Nutr Rev. 1965 Nov 23 (11): 338-40; Kalant H, Mons W, Mahon MA, Acute effects of ethanol on tissue electrolytes in the rat. Can J, Physiol. Pharmacol. 1966 Jan, 44 (1): 1-12.) .

At the end of therapy, mortality of animals in the groups was assessed for 24 and 48 hours, and intoxication markers (activity of enzymes of alanine aminotransferase, levels of glucose, lactate, pyruvate, lipid peroxidation products, diene conjugates, ketodienes of conjugated trienes and Schiff bases were determined in the blood of survivors.

The death of animals occurred during the first 2 days and was associated with hypothermia, collapse and general intoxication, provided the animal was in an unconscious (coma) state for a long time.

So, in the control group, on the first day, 3 rats died (20%), which after the induction of ethanol coma did not go out of the pathological state and did not show a reflex response to pain and light stimuli. Then, during the second day, 3 more animals died with signs of acute fulminant hepatitis with hyperthermia ("marble" liver when opening the abdominal cavity), with a total mortality rate of 40%.

On the first day, the best effect was observed with the introduction of the inventive solution in doses of 20 and 30 ml / kg - in groups of animals there was no death. In groups of animals receiving the claimed solution in doses of 10 (low therapeutic dose) and 40 mg / kg (overload of circulating blood volume), one animal died (6, 7%), as in the group receiving the prototype (6, 7%) .

On the second day of the experiment, deterioration of rats was revealed in all experimental groups. In groups of animals that received the claimed solution in doses of 10, 20, 30 and 40 mg / kg, one individual died, with a total mortality rate of 13.3%, 6.7%, 6.7%, 13.3%, respectively , in the group receiving the prototype, two more individuals (20.0%) (table. 7).

Positive changes during prototype therapy were expressed in normalization of alanine aminotransferase activity and glucose level. The activity of aspartate aminotransferase, the level of lactic and pyruvic acids, however, remained elevated and significantly different in their values from those in rats from the positive control group (Table 8).

When analyzing the dose dependence of the effects of the claimed solution, a positive effect was revealed on all the main indicators. The transition from a dose of 10 ml / kg to a dose of 20 and 30 ml / kg of the claimed solution gave a significant leap in the form of an increase in the positive effect on transaminase activity and energy metabolism (Table 8). At the same time, in their effectiveness, the doses of 20 and 30 ml / kg were almost equivalent and the average therapeutic dose was within the specified range (calculated average dose of 25 ml / kg).

Lipoperoxidation indices (Table 9) against the background of acute ethanol poisoning changed mainly in the form of an increase in the level of Schiff bases, the products of the interaction of aldehydes with amino groups of proteins and amino acids. Moreover, the claimed infusion solution in a dose of 30 ml / kg exceeds the prototype in terms of effectiveness.

Experience 6. The effectiveness of the proposed infusion solution on the model of acute poisoning with a cytostatic cyclophosphamide

The detoxifying effect of the solutions was studied on the model of toxicosis induced by cyclophosphamide in female mice weighing 20-22 g of the BDF1 hybrid line. The studied drugs were administered within 14 days after exposure to cyclophosphamide.

To assess the detoxifying effect of the claimed solution, 5 experimental groups of animals were formed: group 1 (n = 50) of the animals was once intravenously administered cyclophosphamide in a cytotoxic dose of 450 mg / kg (LD ₅₀ ), group 2 (n = 50) and group 3 (n = 50 ) the animals were injected intraperitoneally with the claimed solution at a daily dose of 40 ml / kg and 20 ml / kg, respectively, group 4 (n = 50); the animals were injected intraperitoneally with a prototype at a dose of 20 ml / kg for 14 days; Group 5 (n = 9) - intact animals (Chave Cytotoxic effects of the cyclophosphamide. An experimental study. Arch Ital Patol Clin Tumori. 1967 Jul-Dec; 10 (3): 223-34; Koss LG, Lavin P, Effectsof a single dose of cyclophosphamide on various organs in the rat. 3. Electron microscopic study of the liver, Am. J. Pathol. 1971 Feb, 62 (2): 159-68).

The death of animals and the biochemical blood parameters of the surviving animals were recorded. The material was taken on the 3rd, 7th, and 14th day after the introduction of cyclophosphamide. A biochemical blood test included determination of the activity of aspartate aminotransferase and alanine aminotransferase, the concentration of urea, creatinine, and the content of total bilirubin in the blood serum of experimental animals. In cyclophosphamide, hepatotoxicity is most pronounced, characterized by dystrophic and destructive changes in the liver.

The introduction of cyclophosphamide (group 1) led to 45% death of animals, the death of animals began from 4 days after exposure and continued for 5 days (table. 10).

The introduction of the inventive solution in daily doses of 40 ml / kg and 20 ml / kg against the background of cytostatic administration contributed to a statistically significant decrease in the toxic effects of cyclophosphamide - the death of animals in these animal groups was 10% and 5%, respectively.

In the 3rd group, where a prototype was used against the background of cyclophosphamide administration, there was a 35% mortality rate caused by the toxic effect of cyclophosphamide (Table 10), that is, according to the integral indicator “animal death”, the claimed solution shows a more pronounced detoxification ability than the prototype.

Analysis of laboratory blood parameters of experimental animals (group 1) showed that hepatotoxicity is leading in the toxic effect of cyclophosphamide. During the 7 days of observation after the administration of cyclophosphamide in animals, a statistically significant increase in the activity of the alanine aminotransferase enzyme was observed, and on the 3rd day of observation, a statistically significant increase in the activity of aspartate aminotransferase and the concentration of total bilirubin (Table 11) relative to indices in intact animals. It is characteristic that the transient increase on the 3rd day of aspartate aminotransferase activity also changed on the 7th day with a statistically significant decrease in enzyme activity compared with the level in intact animals. This is apparently due to the development in animals of liver damage by the type of cytolysis.

The results of biochemical studies confirmed a statistically significant decrease in the intensity of toxic reactions of cyclophosphamide with the introduction of the inventive solution in daily doses of 40 ml / kg and 20 ml / kg. Its use leads to the relief of the hepatotoxic effect of cyclophosphamide: the values of biochemical parameters remained at a normal level during the entire time of observation of animals with the introduction of the inventive solution in both doses. For example, with the introduction of the inventive solution in a daily dose of 40 ml / kg on the 3rd day after the administration of the cytostatic agent, the activity of the enzymes aspartate aminotransferase and alanine aminotransferase was 138 ± 21 and 40 ± 4 IU / L, respectively, and with the introduction of the inventive solution in a daily dose of 20 ml / kg - 146 ± 17 and 40 ± 5 IU / l, respectively, while in the cyclophosphamide group - 219 ± 14 and 326 ± 33 IU / l (normal: aspartate aminotransferase - 136 ± 5 IU / l, alanine aminotransferase - 42 ± 2 IU / l). A similar picture was observed on the 7th day of observation.

The concentration of total bilirubin also remained at a normal level, in contrast to the indicator in animals that received only cyclophosphamide, in which on the 3rd day the concentration was 13.1 ± 1.4 μmol / L (normal 6.4 ± 1.0 μmol / L table 11).

The effectiveness of the prototype was also expressed in maintaining the activity of the enzymes alanine aminotransferase and aspartate aminotransferase and the concentration of total bilirubin at a normal level for all periods of observation.

Thus, the detoxifying activity of the inventive infusion solution in relation to cyclophosphamide in various doses was not inferior to the prototype, while there was a significant difference in mortality in the groups of application of the inventive solution in various doses (10% and 5%) and prototype (35%).

The experiments performed show the wide possibilities when using the inventive infusion solution as a detoxifying drug due to the use of a new biologically active component - sodium L-arginine succinate of formula (1).

The inventive balanced infusion solution contributes to the rapid reduction of hypoxic processes in organs and tissues, the removal of under-oxidized waste products of cells, activation of energy metabolism, normalization of water-electrolyte balance and acid-base balance of all body fluids.

In addition, a balanced infusion solution significantly reduces the alkalization of blood and urine, characteristic of the prototype, due to the lower pH of sodium L-arginine succinate, which seems to be advantageous for certain groups of patients.

Thus, the inventive balanced infusion solution has increased detoxifying activity, low toxicity and can be used in the treatment of a wide range of diseases and conditions associated with intoxication of the body of varying severity.

Claims (1)

A balanced infusion solution containing sodium, potassium and magnesium chlorides, a solvent and a biologically active compound, characterized in that it contains sodium L-arginine succinate of the formula as a biologically active component:

in the following ratio of components, wt.%:
Sodium chloride 0.540-0.600 Magnesium Chloride Hexahydrate 0.015-0.030 Potassium chloride 0,025-0,040 Sodium L-Arginine Succinate 1,400-1,700 Water for injections Rest