RU2276974C2 - Medicinal agent "gastropek" for improving digestion process and method for its using - Google Patents

Abstract

FIELD: medicine, pharmacology, pharmacy.

SUBSTANCE: invention relates to the development of a new medicinal agent representing the complex enzyme preparation with the digestive effect. Agent comprises enzyme pectate lyase providing effective cleavage of vegetable food intercellular substances, in particular, pectin and protopectin that results to formation of the homogenous and easily digestible mass, and promotes to assimilation vegetable food that is not assimilated by human organism that results to the more complete digestion of food, its cleavage and assimilation of nutrient substances. The optimal content of enzyme pectate lyase and pancreatin in a tablet covered by the enterosoluble envelope promotes to the more rapid and complete digestion of food in intestine and shows significant advantages as compared with the known digestive preparations.

EFFECT: improved and valuable properties of agent.

5 cl, 11 tbl

Description

The invention relates to medicine, in particular to medicines used for the treatment and prevention of diseases associated with disorders of the digestive process (3). The main indications for the use of drugs of digestive enzymes are: chronic pancreatitis with exocrine insufficiency; pancreatectomy, pancreatic cancer, liver and biliary tract diseases, resection of the stomach, small intestine, chronic enteritis, gastritis with secretory insufficiency, old age, diet errors when eating fatty, unusual or indigestible food, etc.

It is known that Pancreatin is used to treat these diseases (9), which compensates for the deficiency of pancreatic enzymes, contains excretory pancreatic enzymes, lipase, amylase, protease, trypsin, chymotrypsin, promotes the breakdown of proteins (to amino acids), fats (to glycerol and fatty acids) and starch (before dextrins and monosaccharides) and normalizes digestion processes. Pancreatin is present in both domestic and foreign dosage forms, and its content varies widely. So, Pancurmen (Germany) - contains 35 mg of pancreatin; Pancreoflat (Germany) - 170 mg; Festal N (Germany) and Enzistal (India) - 192 mg each; Digestal (USA) - 200 mg; Pancreon 10000 (Germany) - 250 mg; Creon (France) - 300 mg. Domestic drug Pancreatin contains 250 mg. It is known that in addition to pancreatin, some drugs, such as Digestal, Ipental, Panzinorm, Tagestal, Festal, Cholenzym and Enzistal, include bile components that provide emulsification of fats. Vigeratin tablets include pancreatin and lyophilized liver extract; in Panzinorm - cholic acid, amino acid hydrochlorides; in Pancurmen - turmeric extract; in Zimoplex - Dimethicone. It is known that the drugs Abomin, Digestal, Ipental, Tagestal, Festal and Enzistal additionally contain cellulase and hemicellulase, which contribute to the breakdown of plant fiber (6, 7). However, the presence of only cellulases in drugs cannot ensure complete digestion of plant foods and pectolytic enzymes such as pectinases and pectatliases are also necessary for its digestion.

Analysis of drugs used for digestive disorders showed that there is not a single drug manufactured commercially that contains pectolytic enzymes that can break down pectin and protopectin of plant tissue of fruits and vegetables (6, 8).

One of the analogues to the claimed invention is patent No. 1375857, United Kingdom, November 27, 1974, the priority of Germany on November 7, 1970 No. 2053948, "An enzyme that destroys plant material, the process of its production and use in digestion formulations" (4), essential the disadvantage of which is that the enzyme acts only on the plant portion of the food consumed. As a result, additional medication, such as Pancreatin, Festal, etc., will be required. for the digestion of proteins, fats and starchy carbohydrates. Another disadvantage of this invention is the significant mass of the enzyme per unit dose - up to 200 mg, while in the claimed invention the dose of the enzyme pectatliase is 25 mg. In addition, this enzyme is not industrially produced, therefore, it is not possible to compare its macerating properties with the claimed preparation.

The closest prototype in terms of technical nature and the achieved result to the claimed drug Gastropek is Pancreatin and its synonyms - Creon, Mezim-forte, Pancreon 10000, Pancitrat. A significant drawback of these drugs is that they do not fully enough contribute to the digestion of food of plant origin, since pancreatin enzymes are not able to break down pectin and protopectin, which form the basis of intercellular substances and "cement" plant tissue. This function is carried out by pectolytic enzymes, for example pectatliase.

The aim of the invention is the creation of a new drug with a wide spectrum of action that promotes the digestion and assimilation of food of both animal and plant origin, and a method for its use for the treatment and prevention of diseases associated with disorders of the digestive process, which, unlike all currently available digestive preparations with a high activity of the enzyme pectatliase, which ensures the effective breakdown of pectin and protopectin of plant foods; and a full range of Pancreatin enzymes for the breakdown of proteins, fats and starchy carbohydrates.

The purpose of the invention is achieved by the development of the drug Gastropec, consisting of the original substance Gastrolyase, containing the enzyme pectatliase, and the substance Pancreatin, containing the enzymes - protease, amylase and lipase.

The substance of Gastroliasis is a purified enzyme, pectatliase (4.2.2.2.) And has the ability to macerate, i.e. split intercellular substances of plant tissue in a slightly alkaline pH zone for a short period of time (2-4 hours). The pectatliase enzyme was obtained from the culture fluid produced by Bacillus macerans BS-04, grown by deep cultivation by aeration on a specially developed medium. The molecular weight of the pectatliase enzyme determined by gel filtration is 41000-42000 D. The isoelectric point is determined by isoelectric focusing and is in the pH range of 9.4, pectatliase is stable in the pH range of 6.5-8.5, it exhibits optimal specific activity in zone 7 , 5-8.2 at a temperature of 35-40 ° C and is activated by calcium ions.

As a result of the combination of Pancreatin and Gastroliase in one tablet, at their optimal content, the drug Gastropec with a wide spectrum of action was created, which facilitates the digestion and absorption of both proteins, fats, starch, and non-starch polysaccharides of plant foods.

The enzyme pectatliase by maceration of plant tissue provides greater accessibility of the components of the food consumed for the action of pancreatic enzymes, which is confirmed by the data obtained by studying the specific activity of the gastropec preparation in vitro.

Man and many other omnivores are not able to digest the cell walls of plants due to the fact that they do not have the anatomical and physiological capabilities that herbivores have. Most plant foods are coarse-fibrous mass, which has no nutritional value, as it is not absorbed during digestion. Therefore, the use of pectatliase in the composition of the proposed drug is appropriate and leads to the breakdown of pectin and protopectin of plant foods to unsaturated di- and trigalacturonic acids.

In order to determine the dosage of the substance of gastrolyase, the specificity of its effect on plant substrates was studied. When determining specificity, they were guided by the Methodological Recommendations for the experimental study of new enzyme preparations proposed for clinical trials (1). Used a variety of types of plant substrates, the most commonly used in the diet. Evaluation of the effect of the substance Gastroliases was carried out according to the degree of maceration (in%) of these plant substrates, which was expressed in a decrease in the solids content in the substrate due to the release of soluble easily assimilable compounds from plant tissue cells. In addition, we determined the accumulation of degradation products of pectin substances, which turned into a soluble form and are unsaturated galacturonic acids, by spectrophotometry of their optical density in quartz cuvettes with a light-absorbing layer 1 cm thick at a wavelength of 235 nm (2).

As plant substrates, fresh and boiled vegetables and fruits in crushed form were used. To determine the specific action, a standard gastrolyase drug with pectatliase activity of 3,000,000 units / g was used. To establish the minimum amount of gastroliase required for cleavage of the substrate, various amounts of the enzyme from 1000 to 12000 units were studied. per 20 g of substrate.

Table 1.
The specificity of the action of gastroliases on fresh vegetables No. p / p Substrate Amount of enzyme The degree of maceration,% Pectin hydrolysis products
(ΔD) Name Mass g mg units one CARROT 20.0 control - - 0 0 2 20,0 0.3 1000 11.00 3.40 3 20,0 1,0 3000 16.00 7.00 four 20,0 2.0 6000 20.00 10.80 5 20,0 3.0 9000 30.10 13.50 6 20,0 4.0 12000 30,60 13.90 one BEET 20.0 control - - 0 0 2 20,0 0.3 1000 7.10 0.98 3 20,0 1,0 3000 15.60 3.34 four 20,0 2.0 6000 22.90 9.00 5 20,0 3.0 9000 31,50 15,80 6 20,0 4.0 12000 32,10 16.00 one CABBAGE 20.0 control - - 0 0 2 white-headed 20,0 0.3 1000 8.00 3.90 one 20,0 1,0 3000 18.60 8.40 four 20,0 2.0 6000 28,20 10.00 5 20,0 3.0 9000 32,20 13.80 6 20,0 4.0 12000 33.10 14.10 one TOMATOES 20.0 control - - 0 0 2 20,0 0.3 1000 10.00 4,50 3 20,0 1,0 3000 20.00 12.00 four 20,0 2.0 6000 28.00 14.00 5 20,0 3.0 9000 36.00 16.70 6 20,0 4.0 12000 38.00 17,2

The examples in Table 1 show that the maximum percentage of maceration is set when exposed to 9000 units. pectatliase activity (3 mg of gastrolyase with activity of 3,000,000 units / g) per 20 g of fresh substrate. A further increase in the number of gastroliases increases the percentage of maceration slightly and is impractical.

Table 2.
The specificity of the action of gastroliases on boiled vegetables No. p / p Substrate Amount of enzyme The degree of maceration,% Pectin hydrolysis products
substances ΔD Name Mass g mg units one CARROT 20.0 control - - 0 0 2 20,0 0.3 1000 6.0 0.80 3 20,0 1,0 3000 10.5 1.20 four 20,0 2.0 6000 15.0 6.40 5 20,0 3.0 9000 27,2 12.00 6 20,0 4.0 12000 27.9 12.60 one BEET 20.0 control - - 0 0 2 20,0 0.3 1000 6.37 0.30 3 20,0 1,0 3000 9.22 0.92 four 20,0 2.0 6000 19.3 8.00 5 20,0 3.0 9000 28.6 13.00 6 20,0 4.0 12000 30.6 13.60 one CABBAGE 20.0 control - - 0 0 2 white-headed 20,0 0.3 1000 5,0 4.40 3 20,0 1,0 3000 15.6 6.60 four 20,0 2.0 6000 20,2 8.00 5 20,0 3.0 9000 28.1 12,20 6 20,0 4.0 12000 28.5 13.50 one POTATOES 20.0 control - - 0 0 2 20,0 0.3 1000 21.0 0.2 3 20,0 1,0 3000 32,0 1.3 four 20,0 2.0 6000 40,0 2,3 5 20,0 3.0 9000 43.0 2,5 6 20,0 4.0 12000 43.6 2.98 one MIXTURE
VEGETABLES: 20.0 control - - 0 0 3 20,0 0.3 1000 26.0 4,50 3 potatoes - 7 g 20,0 1,0 3000 35,2 6.90 four cabbage - 6 g 20,0 2.0 6000 41,4 8.01 5 carrots - 3 g 20,0 3.0 9000 46.6 10,20 6 beets - 4 g 20,0 4.0 12000 47.9 11.5

The examples in Table 2 show that during the processing of boiled vegetables, such as cabbage, carrots, beets, the degree of maceration is 9-13% lower than on fresh ones, since during the heat treatment of the substrates they partially hydrolyze and some substances pass into a soluble form . The highest degree of maceration among the boiled substrates was found on potatoes, since starch is partially hydrolyzed during heat treatment of potatoes, the content of which reaches 17-20%. This is also evidenced by the low accumulation of hydrolysis products of pectin substances (unsaturated galacturonic acids), the content of which in the potato supernatant is much lower than in the case of other substrates. This explains the fact that the total solids content in the substrates during the preliminary heat treatment is reduced. It was found that the maximum maceration of boiled substrates, as well as under the action of the enzyme on fresh substrates, is observed at a ratio of 3 mg (9000 units) of gastroliases per 20 g of substrate.

Table 3.
The specificity of the action of gastroliases on fresh fruits No. p / p Substrate Amount of enzyme The degree of maceration,% Pectin hydrolysis products
substances ΔD Name Mass g mg units one PEAR 20.0 control - - 0 0 2 20,0 0.3 1000 20,4 6.4 3 20,0 1,0 3000 25.0 10.1 four 20,0 2.0 6000 30.1 14.2 5 20,0 3.0 9000 39.0 19.5 6 20,0 4.0 12000 39.8 20.8 one AN APPLE 20.0 control - - 0 0 3 20,0 0.3 1000 21,2 9.8 3 20,0 1,0 3000 28,2 14.3 four 20,0 2.0 6000 36.6 16.8 5 20,0 3.0 9000 45.9 22.3 6 20,0 4.0 12000 46.2 24.3 one PLUM 20.0 control - - 0 0 2 20,0 0.3 1000 25.0 5.8 3 20,0 1,0 3000 35.6 9.9 four 20,0 2.0 6000 40,2 15.0 5 20,0 3.0 9000 46.1 18.9 6 20,0 4.0 12000 47.1 21.1

The examples in Table 3 show that under the influence of Gastrolyase, fresh fruits increase the degree of maceration compared to fresh vegetables by an average of 10%, since the content of pectin and insoluble protopectin in fruits is higher (5). It was found that the maximum maceration of fruits and vegetables, both boiled and fresh, is observed under the action of 3 mg (9000 units) of gastroliase per 20 g of substrate. It was found that the content of the products of the hydrolysis of pectin substances increases with increasing concentration of the enzyme and reaches a maximum at a concentration of gastrolyase of 3 mg (9000 units) per 20 g of substrate, which is consistent with the degree of maceration of the substrates.

According to the Institute of Nutrition, the amount of plant food a person consumes per day is at least 500 g, so this amount will require 75 mg (with an activity of 3,000,000 units / g) - 225,000 units. Gastroliases, i.e. 25 mg three times a day.

It has been established that the optimal conditions for the action of gastroliase are a slightly alkaline pH zone and a temperature of 37 ° C, which corresponds to the conditions of the small intestine, where the processes of basic digestion and assimilation of food occur. When food enters the stomach, it undergoes acid hydrolysis under the influence of gastric juice; therefore, the effect of gastroliase on substrates pretreated with gastric juice was studied. The substrates, after two-hour exposure in EKVIN gastric juice at a temperature of 37 ° C and stirring, were made alkaline to pH 8.0 and exposed to gastroliases for 6 hours. Studies were performed on both fresh and boiled vegetables. Examples are given in table. 4 and 5.

Table 4.
The specificity of the action of gastroliases on fresh substrates treated with gastric juice No. p / p Substrate Amount of enzyme The number of stomachs. juice, ml The degree of maceration,% Pectin hydrolysis products,
ΔD Name Weight mg units one CARROT 20.0 control - - - - - 7 20,0 - - fifteen 11,4 0.5 3 20,0 3.0 9000 fifteen 42.1 12.9 one BEET 20.0 control - - - - - 2 20,0 - - fifteen 15.3 0.76 3 20,0 3.0 9000 fifteen 41.6 14.2 one CABBAGE 20.0 control - - - - - 2 white-headed 20,0 fifteen 9.6 0.45 3 20,0 3.0 9000 fifteen 43,4 13.9 one MIXTURE 20.0 control - - - - 2 VEGETABLES: 20,0 - - fifteen 12.6 0.62 3 cabbage - 8 g 20,0 3.0 9000 fifteen 44,2 14.6 carrots - 4 g beets - 2 g tomatoes - 6 g one PEAR 20.0 control - - - - - 2 20,0 - - fifteen 9,4 1,2 3 20,0 3.0 9000 fifteen 45.6 20.1 one AN APPLE 20.0 control - - - - - 2 20,0 - - fifteen 11.8 1,5 3 20,0 3.0 9000 fifteen 56.8 22.4

Table 5.
The specificity of the action of gastroliases on boiled vegetables processed by gastric juice No. p / p Name of substrate The mass of the substrate, g Amount of enzyme The number of stomachs. juice, ml The degree of maceration,% Hydrolysis products of pectin-x substances, ΔD mg units one CARROT 20.0 Control - - - - - 2 20,0 - - fifteen 13.3 0.45 3 20,0 3.0 9000 fifteen 38.45 11.9 one BEET 20.0 Control - - - - - 2 20,0 - - fifteen 12.6 0.62 3 20,0 3.0 9000 fifteen 42.0 14.1 one CABBAGE 20.0 Control - - - - - 2 white-headed 20,0 - - fifteen 9.6 0.55 3 20,0 3.0 9000 fifteen 38.8 12.1 one POTATOES 20.0 Control - - - - - 2 20,0 - - fifteen 23,4 0.2 3 20,0 3.0 9000 fifteen 59,4 2.6 one MIX OF VEGETABLES: 20.0 Control 2 potatoes - 7 g 20,0 - - fifteen 24.1 0.4 3 cabbage - 5 g 20,0 3.0 9000 fifteen 54.9 12.6 carrots - 3 g beets - 5 g

From the examples in tables 4 and 5 it is seen that due to acid hydrolysis from 9 to 15% of dry substances passes into the soluble phase. The highest percentage of acid hydrolysis (up to 23%) is observed in potatoes, since starch, the content of which in the composition of potatoes is 15-20%, is more actively hydrolyzed by acid in gastric juice, unlike other polysaccharides of plant substrates, such as cellulose, hemicellulose, glucans and pectins. Subsequent processing of boiled vegetables with Gastroliazi increases the degree of maceration from 38% to 42%, and on potatoes up to 59.4%. Partial acid hydrolysis of substrates by gastric juice intensifies the process of digestion of plant foods by gastrolyase.

Due to the fact that the claimed drug contains two substances: Gastrolyazu and Pancreatinum, studies have been conducted but their compatibility.

Since the content of pancreatin and known medicines ranges from 0.192 g (Festal) to 0.2 g (Digestal, Panzinorm) in one tablet, therefore, its average content was taken as 0.2 g. The gastroliase content in one tablet was set to 0.025 g.

To study the compatibility of the pancreatin and gastroliase enzyme complex, 0.2 g of pancreatin and 0.025 g of gastroliase were mixed. The mixture was dissolved in 100 ml of a phosphate buffer solution with a pH of 8.0 and kept at a temperature of 37 ± 0.5 ° C for 6 hours. Then, the residual activity of the enzymes was determined. Pectatliase activity was determined by spectrophotometry of unsaturated pectolysis reaction products having a maximum optical density at a wavelength of 235 nm (2), the activity of amylase, protease and Pancreatin lipase was determined in accordance with FS 42-3647-98 (9). Examples are given in table.6.

Table 6.
The dynamics of compatibility of the complex of enzymes Pancreatin and Gastroliase Enzyme preparations Gastroliasis Pancreatin After 6 hours of incubation Name Amount mg Activity, units Activity, units Pectatliase Amylase Protease Lipase Pectatliase Amylase Protease Lipase Gastroliasis 25 75,000 70800 Pancreatin 200 - 1500 40 130.0 - 1450 38,2 124.5 Gastroliasis 25 75,000 70200 + + Pancreatin 200 1500 40 130.0 1435 37.6 123,2

From the examples of Table 6 it is seen that the enzymes Pancreatin and Gastroliase do not have an inhibitory effect on each other's activities.

The specificity of the action of the composition of Gastroliase with Pancreatin was studied on mixtures of plant substrates. The amount of Pancreatin was calculated in accordance with the Instructions for use - 1-2 tablets 3 times a day (the content of Pancreatin in 1 tablet is 0.1 g), per day (from 0.3 g to 0.6 g). Therefore, from 20 mg to 24 mg of Pancreatinum and 3 mg of Gastroliase were used for 20 g of plant food.

Experiment progress: A substrate, gastric juice were introduced into six beakers and kept at a temperature of 37 ° C and shaken at a speed of 60 rpm for 2 hours. Then, the pH of the substrates was adjusted to 8.0; made certain amounts of gastrolyase and pancreatin dissolved in 0.001 M phosphate buffer (Table 7), incubated at 37 ° C and shaken at a speed of 60 rpm for 6 hours

Table 7 No.
glasses Weight
substrate, g Volume ml Weight mg gastric juice phosphate buffer Gastroliasis Pancreatin one 20.0 (control) fifteen fifteen - - 2 20,0 fifteen fifteen 3.0 - 3 20,0 fifteen fifteen - 12.0 four 20,0 fifteen fifteen - 24.0 5 20,0 fifteen fifteen 3.0 12.0 6 20,0 fifteen fifteen 3.0 24.0

To compare the effect of the action of enzymes, the substrates were similarly treated separately with Gastrolyase and Pancreatin. Used a mixture of vegetables, both boiled and fresh, in the composition, eaten in the form of salads. Examples are given in table 8.

Table 8.
The specificity of the action of the composition of gastroliase with pancreatin Experience number Substrate Substances The degree of maceration,% The products of the hydrolysis of pectin in-in, ΔD Name Mass g Touring Pancreatin, mg mg units one Mix fresh
of vegetables 20.0 control - - - - - 20,0 3.0 9000 - 45.10 15.60 Cabbage - 6 g 20,0 - - 12.0 4.90 0.25 Carrots - 6 g 20,0 - - 24.0 7.60 0.37 Apples - 5 g 20,0 3.0 9000 12.0 49.4 17.4 Onion turnip - 3 g 20,0 3.0 9000 24.0 56.50 18.3 2 A mixture of boiled
of vegetables 20.0 control - - - - - 20,0 3.0 9000 - 43.0 12,4 Potato - 8 g 20,0 - - 12.0 27.9 1.8 Carrots - 4 g 20,0 - - 24.0 29.6 2,4 Beets - 4 g 20,0 3.0 9000 12.0 59.3 22.6 Cabbage - 4 g 20,0 3.0 9000 24.0 68.3 24.2 3 A mixture of boiled
of vegetables 20.0 control - - - - - 20,0 3.0 9000 - 45.0 11.6 Potato - 5 g 20,0 - - 12.0 22.9 1,2 Carrots - 5 g 20,0 - - 24.0 28.1 1.9 Beets - 5 g 20,0 3.0 9000 12.0 62,4 23.7 Cabbage - 5 g 20,0 3.0 9000 24.0 70.1 25.31

From the examples of Table 8, it was found that under the action of Pancreatin on fresh vegetables, the degree of maceration of plant substrates is insignificant and amounts to 4-7%. On a mixture of boiled vegetables containing potatoes (experiment 2, 3, Table 8), the degree of hydrolysis of the substrate reaches 29-54%, which is the result of the action of Pancreatin amylase, which effectively acts on substrates containing starch. The higher the percentage of potato content in a mixture of vegetables, the higher the percentage of hydrolysis of substrates.

The study of the specificity of the action of the composition of Gastrolyase with Pancreatin on plant substrates pre-treated with gastric juice, showed the effectiveness of the composition. The combined action of Gastroliase with Pancreatin provides a more efficient hydrolysis of plant substrates due to maceration of plant tissues by Gastroliase, as a result of which the substrates become more accessible for the action of Pancreatin enzymes. Thus, the effect of the composition of the substances of gastrolyase with pancreatin is more effective than their action separately.

When developing the drug Gastropec, the property of the substances Gastrolyase and Pancreatin was inactivated in an acidic environment was taken into account, therefore, to protect the core of the tablet of the claimed drug from the action of gastric juice, known excipients and an enteric-film coating protecting the tablet in the pH range 1.2-6.0 were used and maintaining its activity in the pH zone of 7.0-8.5, corresponding to intestinal pH values.

The composition of the tablet Gastropec: Gastroliase - 0.025 g, containing at least 70,000 units. pectatliase activity: Pancreatin - 0.200 g. containing at least 36 units. proteolytic activity: excipients (microcrystalline cellulose, lactose, corn starch, sodium chloride, hydroxypropyl methylcellulose, magnesium stearate). The mass of the tablet core is 0.35 g; the weight of the tablet coated with the enteric coating is 0.36 g ± 5%.

Appearance: white or white tablets with a grayish gray color, coated with an enteric-soluble shell, biconvex, a homogeneous gray mass is visible in the cross section.

To implement the claimed invention, the specific and general pharmacological activity of Gastropek in vitro was studied.

When studying the specific effect of the drug Gastropec on plant substrates, Pancreatin was used as a comparison drug; Mezim-forte (containing only pancreatin) and Festal; Enzistal (containing pancreatin and the hemicellulase enzyme).

Used crushed plant substrates and mixtures thereof both in fresh and in boiled form.

Experiment progress

150 g of substrate were placed in beakers, 150 ml of EQUIN natural gastric juice was added and kept at a temperature of 37 ± 0.5 ° C and shaken at 60 rpm for 2 hours, simulating the work of the stomach. Then the contents of the glasses were alkalinized to pH 8.0, Gastropek and comparison preparations were added and kept at a temperature of 37 ± 0.5 ° С and shaken at a speed of 60 rpm for 6 hours. After 6 hours, the contents of the glasses were centrifuged at 10,000 rpm. min for 15 min Then the supernatant was drained, and the precipitate was washed twice with distilled water of 50 ml each and centrifuged. The washed precipitate was transferred to boxes previously adjusted to constant weight and dried to constant weight at 105 ° C.

The effect of the drug was evaluated but the decrease in the mass of the substrate (degree of maceration) as a result of exposure to Gastropek and comparison drugs and was expressed as a percentage of the ratio of the experimental sample to the control.

The degree of maceration was calculated by the formula:

Where:

M ₀ - the mass of absolutely dry residue of the substrate of the experimental sample;

M _to - the mass of absolutely dry residue of the substrate of the control sample.

In the supernatant obtained after the first separation of the substrate biomass, the accumulation of the hydrolysis products of pectin and protopectin - unsaturated galacturonic acids was determined by spectrophotometry of their optical density (1). The amount of pectin hydrolysis products was determined by the difference in optical densities (ΔD) of the supernatant of the experimental and control samples. The quantitative content of soluble carbohydrates was also determined in the supernatant (2).

The effect of Gastropec and comparison preparations was investigated on mixtures of boiled and fresh substrates in various compositions. As a comparison drug, the Festal drug was used, which contains both pancreatin enzymes and hemicellulase and, therefore, must hydrolyze the hemicellulose of the plant substrate. Examples are presented in tables 9 and 10.

Table 9.
The action of Gastropek and Festal on fresh vegetables Substrate The name of the drug Amount of drug, tablet The degree of maceration,% The products of the hydrolysis of pectin in-in, ΔD Carbohydrates, g Name Mass g one CARROT 150 The control - 10.10 0 0.99 2 150 Gastropek one 32.86 12.35 8.10 3 150 Gastropek 2 38.75 13.72 8.92 four 150 Festal 2 22.10 0.86 7.22 one BEET 150 The control - 15.0 0 1.42 2 150 Gastropek one 39.22 14.15 9.80 3 150 Gastropek 2 42.71 15.32 10.60 four 150 Festal 2 25.13 0.41 9.12 one CABBAGE 150 The control - 9.0 0 0.84 2 150 Gastropek one 45.62 13.76 4.84 3 150 Gastropek 2 49.51 14.91 5.63 four fifteen Festal 2 26.31 1,060 4.73

From the above examples it is seen that the degree of maceration of boiled vegetables is 7-8% less compared to the degree of maceration of fresh vegetables. A study of various dosages of Gastropek showed that there is no significant difference in the effect of the drug on the degree of maceration when using different concentrations of the drug (1 or 2 tablets). When 2 tablets of Gastropec are introduced, the degree of maceration increases slightly, by only 4-10%, depending on the type of substrate. The examples are presented in table.9; 10 show that the degree of maceration of the plant substrate treated with Festal is 17-20% lower compared to the degree of maceration of the plant substrate under the action of Gastropek. The higher level of maceration of substrates with the drug Gastropec is due to the fact that due to the destruction of intercellular compounds in plant tissue, access to the substrate by Pancreatin enzymes is facilitated, as a result of which the formation of soluble carbohydrates contained in vegetables and fruits in the form of polysaccharides increases. As a result, the amount of carbohydrates that went into solution under the influence of Gastropec is 15-20% higher than under the influence of Festal on a plant substrate.

Table 10.
The action of Gastropek and Festal on a boiled substrate No. p / p Substrate The name of the drug Number of tablets The degree of maceration,% The products of the hydrolysis of pectin in-in, ΔD Carbohydrates, g name weight g one CARROT 150 The control - 12.12 0 1,11 2 150 Gastropek one 31,20 11.80 7.80 3 150 Gastropek 2 32.80 12.52 8.13 four 150 Festal 2 16.80 0.40 7.71 one BEET 150 The control - 11.60 0,0 1.30 2 150 Gastropek one 35.21 15.40 9.13 3 150 Gastropek 2 38.12 16.70 10.10 four 150 Festal 2 19.64 0.70 9.32 one CABBAGE 150 The control - 8.60 0 0.8 2 150 Gastropek one 41.12 14.80 4.02 3 150 Gastropek 2 45.51 15.12 5.31 four 150 Festal 2 24.14 0.72 4.81 one POTATOES 150 The control - 22.16 0 6.30 2 150 Gastropek one 69.86 22.90 23.63 3 150 Gastropek 2 71.34 25.12 24.50 four 150 Festal 2 54.60 0.93 23.13

A comparative study of the action of the drugs of the closest analogues and Gastropek was carried out on mixtures of plant substrates. The examples presented in table 11 show that the greatest effect on the degree of maceration of the substrate is observed when using Gastropec, which reaches 52-54%. In the case of Pancreatin and Mezim-Forte, the degree of maceration is only 6-9%. Under the influence of Festal and Enzistal, plant substrates are split by 23-30% due to hemicellulase. present in these preparations, which hydrolyzes the hemicellulose of plant substrates. Under the action on the boiled substrates of the indicated preparations (mixture 3, Table 11), the patterns of change in the degree of maceration remain the same, in absolute terms, 2-4% lower than when exposed to fresh substrates.

Under the action of preparations on a mixture of boiled substrates containing potatoes (mixture 4, 5; tab. No. 11), the degree of maceration is higher than on a mixture of boiled substrates without potatoes. When using Gastropec, it is 69-75%, depending on the content of potatoes in the mixture. Pancreatin and Mezim-forte hydrolyse the substrate by 45-58%, which is explained by the presence of amylase, which is part of the pancreatin enzyme complex that hydrolyzes starch. Enzistal and Festal preparations due to the presence of hemicellulase in them when exposed to a mixture of substrates with potatoes provide their hydrolysis by 51-61%.

Table 11.
The effect of Gastropec and comparison drugs on a mixture of plant substrates No. p / p Substrate The name of the drug Number of tablets The degree of maceration,% Substrate Hydrolysis Products
(ΔD),% Name Mass g one Raw mix No. 1 150 The control - 0 0 Cabbage 60 g Gastropek 2 56.6 19.12 Carrot 25 g Pancreatin 2 9.1 0.20 Tomatoes 30 g Mezim Forte 2 8.3 0.10 Sweet pepper 35 g Festal 2 27.4 0.35 Eneistal 2 29.2 0.42 2 Raw mix No. 2 150 The control - 0 0 Apple 40 g Gastropek 2 54.8 24.33 Carrot 50 g Pancreatin 2 7.6 0.11 Cabbage 60 g Mezim Forte 2 8.1 0.10 Festal 2 28.6 0.38 Enzistal 2 30,2 0.51 3 Mix No. 3
boiled 150 The control - 0 0 Gastroisk 2 52.8 18.93 Beetroot 30 g Pancreatin 2 8.9 0.90 Carrot 30 g Mezim Forte 2 6.39 0.33 Cabbage 50 g Festal 2 23.6 0.94 Tomatoes 40 g Enzistal 2 25.1 1.01 four Mix No. 4
boiled 150 The control - 0 0 Gastropek 2 69.3 21.51 Potato 35 g Pancreatin 2 47.2 0.52 Beetroot 30 g Mezim Forte 2 45.1 0.51 Carrot 30 g Festal 2 51.3 1,56 Cabbage 50 g Enzistal 53,4 1,60 5 Mix No. 5
boiled 150 The control - 0 0 Gastropsk 2 75.41 22.82 Potato 70 g Pancreatin 2 58.60 0.34 Beetroot 20 g Meeim-forte 2 55,42 0.11 Carrot 30 g Festal 2 61.30 0.90 Cabbage 30 g Enzistal 2 63.40 0.84

A comparative analysis of Gastropec with digestive enzyme preparations (Pancreatin, Festal, Enzistal, Mezim-forte) showed that the effectiveness of the action on plant substrates of Gastropec is 17-25% higher.

The effect of Gastropec on the pectin substances of plant substrates is confirmed by the formation of products of their hydrolysis - unsaturated galacturonic acids, the content of which in the supernatant is 22-24% of the control. In the case of using comparison preparations, the accumulation of hydrolysis products of pectin substances contained in the plant substrate practically does not occur and amounts to only 0.1-1%, which is explained by the presence of an insignificant amount of free galacturonic acids.

The introduction of 2 tablets of the drug Gastropec per 150 g of plant substrate increases the degree of maceration by only 3-5%, compared with the action of 1 tablet on the same amount of substrate. Therefore, with normal nutrition, it is recommended to use Gastropek 1 tablet 3 times a day immediately before or during meals without chewing, or 2 tablets 3 times a day when overeating.

A comparative study of the specificity of the action of Gastropec and the preparations Pancreatin, Mezim-forte, Festal, Enzistal showed that Gastropec provides a more complete cleavage of plant substrates - 18-20% higher compared to the preparations Festal, Enzistal, which contain the enzyme hemicellulase, and 22-25% higher in comparison with preparations containing only pancreatin (Pancreatin, Mezim-forte).

The specific activity of Gastropec drugs in vivo was determined by the degree of assimilation of nutrients and balance by the basic elements of animal metabolism, namely nitrogen, calcium and phosphorus. As a comparison, we used well-known enzyme preparations - Pancreatin and Enzistal. It was found that when taking Gastropec, the absolute value of the increase in body weight is higher than when taking Enzistal and Pancreatin.

A study of the assimilation of nutrients (10) in a group of experimental animals when fed with a normal diet with the introduction of Gastropec showed that the use of Gastropec, Pancreatin and Enzistal increases the percentage of feed absorption in such indicators as: dry matter, organic compounds, protein, fat, fiber and biological energy substances. When Gastropec is introduced to animals, the assimilation coefficient is higher in all indicators compared with the control group of animals and is 6-10% higher than with Pancreatin and Enzistal. The greatest differences are observed in the digestion of fiber by animals of the experimental groups, where the assimilation coefficients when using Gastropec are 17.6% higher than in animals of the control group and 11.5-7.5% higher than when using Pancreatin and Enzistal, respectively.

In the group of experimental animals treated with Gastropek in the optimal dose, the percentage of nitrogen assimilation was 5% and 3.5% higher than these indicators compared with the groups of animals treated with Pancreatin and Enzistal, respectively. Studies on the balance (11) of macroelements such as calcium and phosphorus have shown that all three drugs contribute to a more efficient breakdown of food products and the absorption of calcium and phosphorus. However, when using Gastropec, phosphorus is absorbed by 11.4% more compared with groups of animals that received Pancreatinum and 6.1% compared with Enzistal, and calcium is absorbed by 5.3% and 4.9% more compared to Pancreatin and Enzistalom accordingly.

A stable and reliable over a long time increase in body weight of experimental animals treated with Gastropek, and a significant excess of this indicator compared with those obtained with Pancreatin and Enzistal, allowed us to conclude that the composition of the digestive enzymes Gastropec was more specific.

Thus, Gastronek promotes more efficient and complete digestion of food, its breakdown and, as a result, absorption and assimilation of nutrients than well-known enzymatic drugs. used for digestive disorders.

The preclinical toxicological study of Gastropec was carried out at TOKSOFARM LLC at the experimental base of the Laboratory of Pharmacology and Chemotherapy at the NIINA named after G.F. Gauze RAMP in accordance with the "Guidelines for the experimental (pre-clinical) study of new pharmacological substances", 2000 (12).

Gastropec's acute toxicity studies were performed on SHK mice by the method of Litchfield and Wilcoxon modified by Z. Rott (13). Throughout the observation, the condition and behavior of the experimental animals did not differ from the norm. It was not possible to determine the doses characterizing the toxicity of the preparation, since the introduction of the maximum possible doses did not lead to the death of mice because of the low toxicity. The acute toxicity of Gastropec was also investigated (13) in Wistar rats with a single application to the stomach. When animals were opened on day 30, macroscopic changes in the organs were not found. The study of acute toxicity of Gastropec showed that with a single use of the drug in the maximum possible amounts of lethal doses, it is not possible to achieve, therefore, the drug belongs to the group of drugs with low toxicity. In chronic experiments on rats, the drug was shown to be well tolerated by animals. In all doses studied, the drug does not lead to impaired functions of organs and systems and does not affect the structure of internal organs.

Gastropec chronic toxicity studies were performed on Wistar rats (13). Throughout the experiment, deviations in the behavioral reactions of animals were not observed. In no case were the phenomena of mydriasis and exophthalmia observed. Animals of the experimental groups tolerated the administration of the drug. normally added to body weight.

When studying the peripheral blood of rats in experimental and control groups of animals, no significant differences were noted (14, 15). Changes in the morphological composition of leukocytes were not detected in any of the groups. No changes in the coagulation system were found in any group for any period. Analysis of biochemical blood parameters indicates the absence of signs of intoxication in animals after the use of the drug Gastropek. Indicators such as total protein, glucose, enzymes, and electrolytes practically did not change at all periods of observation. When studying kidney function in experimental groups of rats, diuretic disorders were not detected.

When studying the cardiovascular system of animals (16) receiving Gastropek for 30 days, no abnormalities were found.

The state of the central nervous system of rats was not impaired. It was found that a 30-fold daily administration of the drug Gastropec did not have any effect on the central nervous system of experimental animals.

When determining the mass coefficients of internal organs, there were no significant differences in the experimental and control groups. When opening animals, macroscopic changes in internal organs were not found.

A pathomorphological study of the internal organs of rats was carried out after oral administration of gastropec. No damage to the structure of organs and tissues was found.

A study of the chronic toxicity of the drug Gastropec, administered orally daily for 30 days in single doses of 250 and 500 mg / kg (which is 100 and 200 therapeutic doses for humans) showed that the drug is well tolerated by animals. The drug, applied even in a high dose, totaling 15,000 mg / kg, does not lead to changes in the condition and behavior of animals, did not cause death.

Physiological tests showed that the drug Gastropec does not affect the peripheral blood, does not violate the function of the liver, kidneys, cardiovascular system and central nervous system.

It was established that the drug Gastropec does not have allergenic, anaphylactogenic and immunotoxic properties, does not affect the main links of humoral and cellular immunity.

Gastropec's reproductive toxicity and its effect on generative function were studied in Wistar rats (12). It was found that the drug Gastropec does not affect the generative function of experimental rats.

The study of the embryotoxic properties of Gastropec was carried out in accordance with the recommendations (12) on female Wistar rats. It was found that the introduction of Gastropec does not lead to a violation of the gain in body weight of females in comparison with the control. Gastropec when introduced into the stomach at doses of 250 and 500 mg / kg does not cause an increase in fetal mortality, changes in weight and size of the fruit. A study of the internal organs of the fetus (17) also did not reveal differences from the control in the experimental groups of animals. It was found that the offspring of experimental rats in terms of emotional-motor development does not differ from control. The drug has no embryo- and fetotoxic effects, recorded in the antenatal and postnatal period of development and damaging effect on the generative function of animals when introduced into the stomach in all studied doses.

Thus, the drug Gastropec does not have allergenic, anaphylactogenic and immunotoxic properties, does not affect the main links of humoral and cellular immunity. Teratogenic and embryotoxic effects were not identified.

BIBLIOGRAPHY

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Claims (5)

1. A medicine for improving digestion processes, containing pancreatin, which includes amylase, lipase, trypsin, chymotrypsin, and excipients, which is a tablet coated with an enteric-soluble membrane, characterized in that it additionally contains an enzyme pectatlyase isolated from the culture fluid the bacterial producer Bacillus macerans BS-04, in the following ratio of components per tablet core: Pancreatin 0.200 g and proteolytic activity of at least 36 units, pectatliase 0.025 g and pectatlic activity of at least 70,000 units, excipients up to 0.35 g. 2. The drug according to claim 1, characterized in that the pectatliase enzyme is able to macerate pectin and protopectin of plant foods, facilitating its digestion and assimilation. 3. The drug according to claims 1 and 2, characterized in that the pectatliase enzyme is stable in the pH zone of 6.5-8.5, exhibits optimal specific activity in the pH zone of 7.5-8.2 at a temperature of 35-40 ° C has a molecular weight of 41000D-42500D, an isoelectric point of p19.8 and is activated by calcium ions. 4. The drug according to claims 1 to 3, characterized in that it further includes auxiliary substances: microcrystalline cellulose, lactose, corn starch, sodium chloride, hydroxypropyl methylcellulose, magnesium stearate. 5. A method of improving digestion, including the introduction of an enzyme preparation, characterized in that the drug is administered according to claims 1-4.