RU2225715C1 - Hepatoprotecting agent - Google Patents

Abstract

FIELD: medicine, hepatology, pharmacy. SUBSTANCE: invention relates to the development of hepatoprotecting agent. The agent is made as injection solution and comprises essential phospholipids, benzyl alcohol, dihydroxycholic acid, sodium chloride, sodium hydroxide, riboflavin, α-tocopherol, ethanol and injection water taken in the definite ratio of components. Agent is non-toxic and devoid significant alterations in enzymes activity, hemoglobin level, amount of platelets and leukocytes in administration. EFFECT: valuable medicinal properties of agent. 3 tbl, 2 dwg

Description

The invention relates to medicine and relates to a complex preparation in the form of an injection solution containing essential phospholipids, the active principle of which are diglyceride esters of cholinophosphoric acid and unsaturated fatty acids: linoleic, linolenic and oleic.

The known drug “Essentiale” made in Germany is known (Vidal Handbook, ed. 2. - M., 1996, p. B-816; Mashkovsky MD Medicines, ed. 12. - M., 1993, vol. II, p.52). It is available in the form of capsules and an injection solution in ampoules of 5 ml. In an injectable form, one ampoule contains essential phospholipids 250 mg, pyridoxine hydrochloride 2.5 mg, cyanocobalamin 10 μg, nicotinamide 25 mg and sodium pantothenate 1.5 mg.

The objective of the present invention is the creation on the basis of essential phospholipids of a domestic preparation of hepatoprotective action in an injectable dosage form. Due to the qualitative and quantitative selection of ingredients, the drug is non-toxic; when administered in a therapeutic dose, it does not cause statistically significant changes in the activity of the enzymes alanine, aspartotransaminase and alkaline phosphatase, in the concentration of hemoglobin, in the number of platelets and leukocytes.

To solve this problem, a hepatoprotective agent in the form of a solution for injection containing essential phospholipids, benzyl alcohol, dioxycholic acid, sodium chloride, sodium hydroxide, riboflavin, α-tocopherol, ethanol 96 ° and water for injection is proposed. The tool contains these ingredients in the following ratio, wt.%:

3- (Phosphatidyl) soybean choline,

containing 93% (3-phosphatidyl) choline 4.5-5.5

Benzyl alcohol 0.8-1.0

Dioxycholic acid 2.0-2.6

Sodium chloride 0.2-0.3

Sodium hydroxide 0.2-0.3

Riboflavin 0.009-0.011

α-Tocopherol 0.013-0.02

Ethanol 96 ° 0.25-0.4

Water for injection Else

An example of obtaining an injectable solution of essential phospholipids

One ampoule (5 ml) contains, mg:

3- (Phosphatidyl) soybean choline,

containing 93% (3-phosphatidyl) choline 250

Benzyl alcohol 45

Dioxicholic Acid 115

Sodium Chloride 12

Sodium hydroxide 11.5

Riboflavin 0.5

α-Tocopherol 0.75

Ethanol 96 ° 16.304

Water for injection Up to 5 ml

To prepare 1 l of the drug, 700 ml of water for injection is heated to a temperature of 45 ± 5 ° C and saturated with nitrogen. 2.4 g of sodium chloride are charged and dissolved. Download 23 g of dichycholic acid, after stirring, a white suspension is obtained. To the suspension, with stirring, add a 1 M solution of sodium hydroxide until complete dissolution of the dioxycholic acid, the pH value being 8.8 ± 0.2. The consumption of 1 M sodium hydroxide solution is about 60 ml.

In a clear, colorless solution at a temperature of 45 ± 5 ° C, the calculated amount of essential phospholipids is loaded and mixed until a homogeneous yellow suspension is obtained.

After adding 9 g of benzyl alcohol and stirring, a clear solution of light yellow color is obtained.

Add 0.1 g of riboflavin and mix until completely dissolved, obtaining a bright yellow solution with greenish opalescence, which is then cooled with stirring to a temperature of 20 ± 2 ° C.

The calculated amount of α-tocopherol is dissolved in 96 ° ethanol and added to the solution obtained with stirring. The volume of the solution was adjusted to 1 liter with water for injection, the solution was stirred and saturated with nitrogen.

After filtration through a Millipor filter with a pore size of 22 μm, a clear solution is obtained, which is subjected to sterilizing filtration and poured into a ampoule of dark glass in a stream of nitrogen.

A comparative study of the proposed drug and the drug “Essential N” - an injection solution in ampoules of 5 ml produced by the company “Ron-Pulenk Rohrer” (Germany), a series of 18541.

The active substance of the drug “Essential N” - essential phospholipids (substance EPL) - are the main elements in the structure of the cell membrane and cellular organelles of the liver. In the body, EPLs have a normalizing effect on the metabolism of lipids, proteins and the detoxification function of the liver; restore and maintain the cellular structure of the liver and phospholipid-dependent enzymatic systems; inhibit the formation of connective tissue in the liver.

“Essential N” is used for fatty liver degeneration of various etiologies, acute and chronic hepatitis, liver cirrhosis, liver coma, radiation syndrome, toxicosis of pregnancy, poisoning, drug and alcohol poisoning of the liver, impaired liver function as a result of complications from other diseases, psoriasis.

The absorption spectrum of the preparations was determined on an Ultrospec-II spectrophotometer manufactured by LKB Biochrom. The preparations were diluted with distilled water in a ratio of 1: 3.5, and the resulting solution was filtered through a Millipor filter with a pore size of 0.45 μm.

Figure 1 presents an overview of the absorption spectrum of a solution of the comparison drug, taken in the wavelength range from 300 to 550 nm. Two pronounced absorption peaks are observed: one at 370 nm (0.72 units of optical density), the other at 443 nm (0.73 units of optical density).

Figure 2 shows the absorption spectrum of a solution of the proposed drug, shot in the same wavelength range. Two absorption peaks are also observed here at the same wavelengths as in FIG.

The spectra of the preparations are similar in location of the peaks of maximum absorption, which indicates the identity of the active substances in the solutions of the preparations.

The acute toxicity of the drugs was determined. To do this, outbred white mice were injected into the tail vein with 0.5 ml of the correspondingly diluted proposed drug or drug comparison. The doses at which the drugs were administered were 400, 500, 600, 700, 800 and 900 mg / kg. Mice were monitored for two weeks after a single injection.

Both the proposed drug and the comparative drug, when administered at doses of 400-500 mg / kg, caused an increase in the respiratory rate, the number of uria and defecation, and the activation of salivation. At doses exceeding 500 mg / kg, the appearance of seizures was observed - first tonic, with a further increase in dose - generalized clonic-tonic. The death of animals occurred due to respiratory arrest.

For the proposed drug, the LD ₅₀ value was 634 ± 65 mg / kg, for the comparison drug 667 ± 70 mg / kg.

Subacute drug toxicity was studied on outbred white rats weighing 180-220 g. Preparations in a volume of 1 ml were injected into the tail vein of animals. Based on the fact that the therapeutic dose of drugs is 40 mg / kg, and a ten-fold therapeutic dose is 400 mg / kg, laboratory animals were divided into five groups:

- group 1 - 10-fold dose of the proposed drug,

- group 2 - therapeutic dose of the proposed drug;

- group 3 - 10-fold dose of the comparison drug;

- group 4 - therapeutic dose of the comparison drug;

- group 5 - intact animals (saline).

The drugs were administered to rats for 14 days. The degree of intoxication was assessed by the general condition of the animals, the condition of the coat, the change in the number of uria, defecation, salivation, the onset of tremor, seizures, changes in the frequency and rhythm of respiration.

After the introduction of drugs, animals were slaughtered and their pathological dissection was performed. Samples of tissue of the kidneys, liver, spleen, thymus and brain were taken for histological analysis and blood samples for determining biochemical and hematological parameters.

Table 1 presents the results of a biochemical blood test of laboratory animals. Between the results obtained for the proposed drug and the comparison drug, no significant difference was observed either with the introduction of the therapeutic dose or with the introduction of a 10-fold dose of the drugs. There is no difference from the data obtained in the control group of animals.

The results of hematological analysis are given in table.2. They practically do not differ in all groups of animals, including the control.

In response to the administration of drugs in both studied doses, the leukocyte blood count of laboratory animals did not change (Table 3).

Histological studies in determining subacute toxicity showed that with the introduction of therapeutic doses of the proposed drug and the comparison drug, no pronounced pathological changes were observed in the studied organs of animals, including the gastric mucosa. The determining characteristics of the microscopic structure of the liver, kidneys, thymus, spleen, and brain tissue corresponded to the norm and were the same as in the control group of animals.

A similar pattern was observed with the introduction of a 10-fold therapeutic dose of the comparison drug.

Studies conducted after the introduction of a 10-fold therapeutic dose of the proposed drug showed that the macroscopic structure of the liver, kidneys, thymus, spleen, brain tissue and gastric mucosa corresponds to those in control animals.

Microscopic studies showed the following:

Liver. Without visible changes, the typical lobation of the structure is clearly expressed. There are single small-cell infiltrates along the portal tracts in the parenchyma. Separate central and sublobular veins are dilated and filled with blood. There are areas with enhanced capillary pattern, endotheliocytes are clearly visible.

The kidneys. In general, the histoarchitectonics of an organ does not differ from that in the control group of animals. The cortical substance is represented by vascular glomeruli and tubules of the nephron. The capsule fits snugly to the organ parenchyma. Large vascular arteries preserved, vascular walls unchanged. The brain substance is represented by the rays of the collecting tubes, longitudinal and transverse sections of the nephron loops. Epithelial lining of the tubules without pathological changes. There are areas with focal small cell infiltrates.

Thymus. Without visible pathological changes detected by the histological method. The cortical substance is covered with a thin capsule and prevails over the cerebral. Mostly small thymocytes are found in the medulla, and the periphery of the cortex is abundantly infiltrated by large and medium thymocytes. Capillaries dilated in places and blood-filled.

Spleen. Of usual structure, the capsule is well developed and contains fibrous components. White pulp is detected in the form of periarterial couplings and primary lymphatic follicles. The border between the red and white pulp is unsharp. Red pulp is enhanced by dilatation of sinusoidal structures and pronounced filling of the red blood cell mass.

Brain. The bark has five distinct layers with cells characteristic in shape and size. The width, cell density of the layers, cytoarchitectonics, the state of the basophilic substance correspond to the norm.

Due to the fact that the solutions of the preparations were administered to the animals in the tail vein, the locally irritating effect of the preparations was evaluated by the status of the tail epithelium. Using a binocular lens, a visual examination of the injection site was carried out in order to detect hematomas, impaired epithelial integrity and the presence of ulcerative lesions. In all groups of laboratory animals, single hematomas were noted, and their number did not depend on the dose of the drug administered.

No other manifestations of the locally irritating effect of the drugs were detected during macroscopic examination.

The histological picture of the prepared tails in the control group of animals is typical. The muscles are stained, the cartilaginous structures are clearly structured, and the vessels are well defined. The veins are moderately dilated.

In groups of animals that were administered the proposed drug and the comparison drug in both doses, the histological picture without pronounced pathology, the cytoarchitectonics of the injection site, is the same as in the control.

Claims (1)

1. Hepatoprotective agent, characterized in that it is a solution for injection and contains 3- (phosphatidyl) choline from soybeans containing 93% 3- (phosphatidyl) choline, benzyl alcohol, dichycholic acid, sodium chloride, sodium hydroxide, riboflavin , α-tocopherol, ethanol 96 ° and water for injection in the following ratio of ingredients, wt.%: 3- (Phosphatidyl) choline from soybeans containing 93% (3-phosphatidyl) choline 4.5-5.5 Benzyl alcohol 0.8-1.0 Dioxycholic acid 2.0-2.6 Sodium chloride 0.2-0.3 Sodium hydroxide 0.2-0.3 Riboflavin 0.009-0.011 α-Tocopherol 0.013-0.02 Ethanol 96 ° 0.25-0.4 Water for injection Else

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