UA140278U - METHOD OF OBTAINING A COMPLEX OF BIOLOGICALLY ACTIVE SUBSTANCES OF DOG NETTLE HERB WITH PHENYLALANINE WITH SEDATIVE AND DIURETIC ACTION - Google Patents

Abstract

The method of obtaining a means with sedative and diuretic effects includes extraction of dog nettle herb with 70% ethyl alcohol solution, purification, filtration, and the extraction is carried out twice in the ratio of raw material to extractant 1: 5-1: 10, adding to the filtrate phenylalanine in triplicate. to the total amount of phenolic compounds by evaporation to dry extract.

Description

The utility model belongs to the pharmaceutical field, namely to a therapeutic and preventive agent based on the herb dog nettle with sedative activity, which is intended for the treatment and prevention of functional disorders of the central nervous system.

Stinging nettle is one of the most widely used medicinal plants with a sedative effect. Galenic agents or dry raw materials are part of many medicines. One of the most common medicines based on this medicinal plant is dog nettle tincture (11.

The method of obtaining tincture of dog nettle is generally known (1, 2, 3). However, the disadvantage of this drug is the instability of the chemical composition, and as a result, the pharmacodynamics.

In addition, in the preparation of the tincture, ethyl alcohol is used as an extractant, which not only limits the number of patients who can use this medicinal product (children, pregnant women, nursing mothers, persons whose activities require increased attention, etc.), but is also capable of affect the central nervous system and modulate the effect of the drug itself. Therefore, aqueous extracts of 3 of these raw materials are recommended for these categories of patients, but these dosage forms are non-standardized, poorly stored and have a different chemical composition.

The basis of the useful model is the task of developing a method of obtaining a new therapeutic and preventive agent from dog nettle grass with a more pronounced sedative and diuretic effect, which does not contain ethanol.

According to a useful model, an alcohol extract is obtained from nettle grass 70 95 with ethanol in a ratio of 1:5-110, the purification of which is carried out by settling and separating the supernatant, after which the amino acid phenylalanine is added in a threefold equimolar amount in relation to the total amount of phenolic compounds. leave for the formation of complexes for a day and evaporate the solvent to obtain a dry extract.

The obtained dry extract from the nettle grass contains at least 7 95 sum of phenolic compounds in terms of gallic acid and not less than 1 90 flavonoids in terms of hyperoside |2, 3, 4).

The claimed remedy is obtained in a simple, affordable way. In addition, the obtained agent in dry form differs advantageously in terms of technological parameters: it is characterized by satisfactory free-flowing properties and lower hygroscopicity, which ensures stability at

From storage.

The claimed product does not contain ethanol, which expands the possibility of its use in children, pregnant and lactating women, and the above categories of persons. It is also possible to add an appropriate amount of the amino acid phenylalanine to the tincture of dog's nettle, and in the subsequent extraction of the extractant, a dry extract with a residual moisture of 4-10 95, which corresponds to the obtained product, can be obtained.

A useful model is illustrated by examples:

Example 1.

350 ml of 7095 ethanol (taking into account the absorption coefficient of the solvent) was added to 50 g of nettle grass, crushed to a particle size of 1-2 mm, which was placed in a flask, and extracted for 5 days at room temperature. The extraction was repeated twice with a new portion of extractant (250 ml). The extracts obtained were combined, stood for a day, and filtered. The dry residue in the tincture was 2.04 95, the content of the sum of phenolic compounds - 8.17 96. 2.5 g of phenylalanine was added to 500 ml of the obtained tincture, which is three times the equimolar amount in relation to the sum of phenolic compounds. The resulting mixture was left to infuse for a day to obtain complexes of phenolic compounds with phenylalanine, after which the solution was filtered and evaporated using a rotary vacuum evaporator to a dry extract. The yield of the dry extract is 22 95. The obtained extract is a greenish-brown powder with a characteristic smell.

According to the results of the phytochemical analysis of the obtained extract by the method of thin-layer chromatography, iridoids and flavonoids were detected. The content of flavonoids in terms of hyperoside |2I in the extract was 1.61--0.04 95.

Example 2.

The pharmacological activity of the obtained extract from the nettle herb and industrially produced nettle tincture (Lubnyfarm OAO, series 062021) was studied in the elevated cruciform maze test (5, 6, 71.

The study was performed on 23 white non-linear mice weighing 18-20 g, randomly selected for the experiment. Animals were kept in standard vivarium conditions

of the Central Research Laboratory of the National Pharmaceutical University on a standard water-food ration with a natural "day-night" lighting regime.

Immediately before administration to the animals, the nettle tincture was prepared as follows: the alcohol was distilled from the nettle herb tincture and brought to the original volume with purified water. The resulting solution was injected intragastrically through a probe at a dose of 5 ml/kg 30 minutes before the test. The dry extract modified by phenylalanine was injected into the stomach in the form of a 1.5 95 aqueous solution at a dose of 5 ml/kg.

The "elevated plus maze" test (EPL) is a screening behavioral test for the anxiolytic activity of pharmacological agents in rodents. It is based on the unconditional reflex of mice and rats to avoid brightly lit unfamiliar spaces and, if possible, to hide in a darkened compartment of the device ("mink reflex"). Therefore, the device consists of two open sectors ("sleeves") measuring 50 x 10 cm (length x width x width) and two crosswise placed closed sleeves measuring 50 x 10 x 10 cm (length x width x height). The maze is stirred at a height of 75 cm. Animals kept in a dark place for 5 minutes are placed on a lighted platform with their heads facing the open sleeve. For

The number of visits to the dark and light arms, as well as the time spent by the animal in the light and dark arms and on the central platform, are recorded in minutes. The results are shown in Table 1.

Table 1

The influence of the studied substances on the performance of mice in the test of the elevated cross-shaped maze

Time, Quantity Time, Quantity Time,

Group conducted in visits conducted in visits conducted dark dark light light on the center. compartment | compartment compartment | compartment site control (IR)

Tincture of dogwood 84,057,97 5,55-1,3 85,5-6,6" 6,050,8 10,5-6,8 nettle

Stinging nettle extract (ESC) with phenylalanine

Note. ESC - dog nettle extract. The differences are statistically significant: "7 - with the control group (reb0.05), 77 - with the control group (reb0.01).

Animals of the intact control (IC) group spent most of the time in darkened compartments (sleeves). The percentage distribution of time (dark arms: light arms: central platform) for the animals of the IR group was 85.4 95: 11.095: 3.690. Such a result generally corresponds to the typical picture of the behavior of rodents in a new unfamiliar space and corresponds to the data of the literature I51I.

The most pronounced under these conditions was the anti-anxiety effect of ESC compositions with phenylalanine, which reduced the time the animals spent in the dark compartments by 72.7 95 (p«0.05 vs. IR). In addition, against the background of the effect of this pharmacological agent, the time the animals spent in the lighted arms increased by 5.8 times (p<0.05 vs. IR). At the same time, the anxiolytic effect of this agent on both indicators (time in the dark and light sleeves) statistically significantly (p<0.05) outweighs the similar effect of the nettle tincture. This may indicate potentiation

The effect of the amino acid phenylalanine.

According to the literature data, the ability of pharmacological agents to reduce the time rodents stay in dark compartments by more than 50-95, as well as to increase the time spent in lighted compartments by 2-4 times, can be considered as an effect inherent to true anxiolytics. Thus, in view of the obtained results, pharmacological compositions based on dog nettle extract and phenylalanine can be considered as promising agents with anti-anxiety activity.

Example 3.

To determine the state of excretory function of the kidneys, test 3 water load is used. To do this, settled water at room temperature (in the amount of 95% of the animal's weight) is injected into the stomach of mice through a probe. 20-25 minutes before the introduction of water into the stomach, the mice are intragastrically injected with the pharmacological agents presented in Table 2. During 4 hours, the animals are observed and the volume of urine released during this time is measured.

Table 2

The effect of the studied substances on the diuresis of mice 4 hours after administration

Note. The differences are statistically significant (p<0.05): " - with the intact control group.

As can be seen from Table 2, the average value of diuresis of animals of the intact control group during 4 hours is 0.93 ml. Such a volume of urine released in rats within 4 hours corresponds to the physiological norm (7). Against the background of water stress, dog nettle extract with phenylalanine increases diuresis in animals during this observation period. Therefore, against its background, the volume of excreted urine statistically significantly increases by 51.6 95, respectively, compared to the indicator of animals of the intact control group (p<0.05).

The obtained results may indicate the presence of diuretic activity in ESC with phenylalanine. It is worth remembering that herbal medicines most often show an integral diuretic effect, which is characterized by an effect on all stages of urination: glomerular filtration, tubular reabsorption, as well as secretion. Therefore, a more detailed study of the effect on urinary excretion of these promising pharmacological agents is necessary.

Thus, a new method of obtaining a drug with sedative and diuretic activity from nettle grass using phenylalanine is claimed, which is characterized by a number of advantages: 1) ease of implementation and the possibility of implementation on standard equipment in industrial conditions of Ukraine; 2) the use of 70-95 ethanol solution as an extractant, that is, the method is available, environmentally safe, cheap, and does not require working with toxic and harmful reagents; 3) the use of dog nettle grass as a raw material, which has a sufficient raw material base, is widely distributed and cultivated on the territory of Ukraine;

From 4) the presence of a pronounced sedative and diuretic activity of a dry extract from the nettle herb obtained by the claimed method; 5) the prospect of using the obtained dry extract as a medicinal substance for the preparation of various medicinal forms; b) the absence of ethanol in the extract obtained by the claimed method allows its long-term use in treatment on the recommendation of a doctor for a wide range of patients, including children; 7) a long period of storage of the product obtained by the declared method.

Sources of information: 1. Mashkovsky M.D. Medicines. Ed. 16th, revised, ex. and additional / M.D.

Mashkovsky. - M.: OBO "Izd-vo Novaya Volna", 2010. - 1216 p. 2. State Pharmacopoeia of Ukraine: in Vol. 3 / SE "Ukrainian Scientific Pharmacopoeia Center for the Quality of Medicinal Products". - 2nd edition. - Kh.: State enterprise "Ukrainian Scientific Pharmacopoeia Center for the Quality of Medicines", 2015. - Vol. 1. - 1128 p. 3. State Pharmacopoeia of Ukraine: in Vol. 3 / SE "Ukrainian Scientific Pharmacopoeia Center for the Quality of Medicinal Products". - 2nd edition. - Kh.: State enterprise "Ukrainian Scientific Pharmacopoeia Center for the Quality of Medicinal Products", 2014. - Vol. 3. - 732 p. 4. Research on the chemical composition of the components of dog nettle grass / E.A. Romanenko, O.M.,

Koshovoy, A.M. Komisarenko, O.I. Golembiovska, Yu.I. Gladysh // Herald of Pharmacy. - Kharkiv, 2018 -

Mo Z (95). - C 34-38.

5. Buresh Ya., Bureshova O., Huston D. Methods and the main experiment! on the study of the brain and behavior. - M.: Viessh. Shk., 1991. - P. 119-122. 6. Ota bizsomegu apa EmaiYpayop: Rnaptasoiodisa! Azzauz / unit Bu N. Modei. Zrhipdeg Zsiepse 8

Vyvzipez55 Media, 2007. - R. 902-942. 1. Nephroprotective properties of primrose preparations / O.O. Koiro, S.Yu. Strigol,

O. Ya. Mishchenko // Water: hygiene and ecology. - 2014. - Mo 1-4 (2). - b. 115-127.

Claims (1)

FORMULA OF THE USEFUL MODEL The method of obtaining a means with a sedative and diuretic effect, which includes the extraction of nettle grass 70 95 with a solution of ethyl alcohol, purification, filtration, which is characterized by the fact that the extraction is carried out twice in the ratio of raw materials to the extractant 1:5-1:10, with the addition of to the filtrate of phenylalanine in a threefold equimolar amount in relation to the total amount of phenolic compounds, by evaporation to a dry extract.