RU2792378C1 - Method of obtaining an agent with diuretic activity - Google Patents

Abstract

FIELD: chemical; pharmaceutical industry.

SUBSTANCE: invention relates to a method for producing an agent with diuretic activity. A method for obtaining an agent with diuretic activity, which consists in biogenic stimulation of fresh herb of the lady's mantle collected in the phase of flowering and the beginning of fruiting, in darkness and cold for 7 days in a sealed package at a temperature of +4°C, followed by grinding to particles of 3-5 mm in size, pressing, canning with 95% ethyl alcohol in the following ratio juice : alcohol - 10:2, keeping the juice at a temperature of +4°C for 10 days, filtering.

EFFECT: above method allows to improve the quality of the product obtained, to obtain an agent with a significant superiority in diuretic activity with a potassium-sparing effect.

1 cl, 2 tbl

Description

The invention relates to the pharmaceutical industry and relates to a method for producing a herbal medicinal product with diuretic activity.

A method for obtaining an aloe extract is known (see patent RU 2125882 pub. 10.02.199), in which the raw material is preserved in the dark and in the cold, followed by extraction by boiling in water.

The disadvantage is the presence of additional production stages - extraction, boiling, which leads to the defermentation of raw materials.

The technical objective of the invention is to expand the arsenal of agents with diuretic activity, improve the quality of the resulting product and simplify the technology.

The technical result of the invention is to improve the quality of the resulting product, a significant superiority in diuretic activity of the agent obtained by the claimed method, compared with the infusion of the common cuff, the diuretic activity of the agent is accompanied by a potassium-sparing effect.

The advantage of the proposed method is the absence of a heating process and, as a result, the preservation of the enzyme system in the raw material with a predominance of peroxidases and catalases, which contribute to the formation of many forms of phenolic compounds that cause a wide range of pharmacological activities, including diuretic [3,7].

The search for new drugs with a diuretic effect is due to the wide spread of diseases of the cardiovascular system and kidney diseases. Diuretics with a potassium-sparing effect have an advantage.

The prognostic assessment of the manifestation of the diuretic activity of the cuff juice was based on the chemical composition. According to the literature data, oxidized forms of flavonoids (flavones, flavonols and their glycosides) together with other groups of biologically active substances (polysaccharides, saponins, terpenes) cause a diuretic effect [6].

The source of juice is the aerial part of the common fresh cuff. At the moment, this type of raw material is little studied, as evidenced by limited publications. The literature available to us describes the preparation of an ethanol extract with antiviral activity [4, 5].

How to obtain the final product:

Common fresh grass, harvested in the phase of flowering and the beginning of fruiting, is subjected to biogenic stimulation, for this it is hermetically packed and placed in a dark, cool place at a temperature of +4 ° C and kept under these conditions for 7 days. After this time, the raw material is crushed to particles with a size of 3-5 mm. The resulting mass is squeezed through 4 layers of gauze and preserved with 95% ethyl alcohol in the ratio of juice:alcohol - 10:2. The juice is kept at a temperature of +4°C for 10 days, after which the juice is purified from the precipitate formed by filtration through a paper filter. The volume of juice obtained is from 42% to 51%, with raw material moisture content from 55% to 84%.

The assessment of diuretic activity was carried out in accordance with the guidelines [1.2]. The study was carried out on white outbred male rats weighing 180–220 g, which were kept under standard vivarium conditions with a natural light regime, at room temperature, without restrictions on access to food (standard, certified compound feed in accordance with current standards) and water. The keeping of animals met the requirements of the Order of the Ministry of Health and Social Development of Russia dated August 23, 2010 No. 708n “On Approval of the Rules of Laboratory Practice” and the “Rules of the European Convention for the Protection of Vertebrate Animals Used for Experimental and Scientific Purposes” (Strasbourg, 1986).

To study the diuretic activity of the tested substance, male rats were divided into four groups of six animals (1 - intact, received drinking water; 2 - experimental received drinking water with oral administration of an ordinary cuff infusion at a dose of 20 mg/kg (in terms of dry residue); 3 - the experimental received drinking water with oral administration of the juice of biostimulated common cuff herb at a dose of 20 mg / kg, in terms of dry residue; 4 - the experimental received drinking water with oral administration of the comparison preparation of wild strawberry infusion at a dose of 20 mg / kg, in terms of dry residue for dry matter). The studied animals were orally administered drinking water in an amount of 5% of body weight, after which they were placed in urinals, urine was taken after 1, 3, and 5 hours [1, 2]. For urine samples, the volume and concentration of sodium and potassium electrolytes were determined by the X-ray fluorescence method, using an energy-dispersive X-ray fluorescence spectrometer of the Thermo Scientific QUANT'X brand. Based on the obtained data, the diuretic index (CI) was calculated as the ratio of the volume of urine in the test sample to the volume of the control and the saluretic index (SI) was calculated as the ratio of the concentration of the element in the test sample to the concentration in the control.

With the introduction of cuff juice from biostimulated raw materials, a statistically significant increase in diuresis was observed after 5 hours in comparison with the intact group, which led to an increase in total urine volume (CI = 2.8). The test substance was significantly superior in diuretic activity to the infusion of common cuff in periods of 1 and 5 hours. The diuretic activity of the tested substance is comparable to that of wild strawberry infusion; the reference drug showed greater activity in the period of 1 - 3 hours, while the juice was more active in the period of 3 - 5 hours (table 1).

It was found that the juice of the biostimulated grass of the common mantle significantly reduced the concentration of sodium and did not change the concentration of potassium. An infusion of herb cuff at a dose of 20 mg/kg significantly reduced the concentration of sodium and caused an increase in potassium in the urine. Comparator infusion of wild strawberry reduced the excretion of sodium and potassium (table 2).

The proposed method allows to obtain a drug with diuretic activity, accompanied by a potassium-sparing effect.

The proposed method is implemented in the following example:

Fresh grass of the common mantle collected in the phase of flowering and the beginning of fruiting weighing 1000.0 was hermetically packed in polyethylene and placed in a cool dark place with a temperature of +4°C for 7 days, after this time the raw material was crushed to particles of 3-5 mm in size, and squeezed out of it through 4 layers of gauze 490 ml of juice. The resulting juice was preserved by adding 82 ml of ethyl alcohol 95%. Purification was carried out by settling in a cool place at a temperature of +4°C for 10 days, the precipitate formed was filtered through a paper filter.

The proposed method for obtaining juice is quite simple, does not require complex technological schemes, allows you to obtain a diuretic with a potassium-sparing effect and expand the range of drugs with the claimed activity.

Literature:

1. Bryukhanov V.M., Zverev Ya.F., Lampatov V.V., Zharikov A.Yu. Methodical approaches to the study of kidney function in animal experiments / Nephrology, 2009 vol.13. No.3, p.52-62.

2. Berkhin, E.B. Methods for studying the effect of new chemical compounds on kidney function / E.B. Berkhin // Chem.-Pharmac. magazine -1977 -T.11, No. 5 -S. 3-11.

3. Kolesnikova, L. I. Free radical oxidation: the view of a pathophysiologist / L. I. Kolesnikova, M. A. Darenskaya, S. I. Kolesnikov // Bulletin of Siberian Medicine. - 2017. - T. 16. - No. 4. - S. 16-29. - DOI 10.20538/1682-0363-2017-4-16-29.

4. Mazurkova N.A., Protsenko M.A., Filippova E.I., Kukushkina T.A., Vysochina G.I., Lobanova I.E., Mazurkov O.Yu., Shishkina L.N., Agafonov A.P. Investigation of the antiviral activity of experimental samples of preparations obtained from the herb and roots of Alchemilla vulgaris L. against vaccinia and mouse pox viruses. Development and registration of medicines. 2019;8(4):9-15.

5. Filippova E. I., Kukushkina T. A., Lobanova I. E., Vysochina G. I.,

Mazurkova N. A. Antiviral properties of the drug based on the amount of flavonoids of the common cuff ( Alchemilla vulgaris L.) in relation to the influenza virus. Basic research. 2015; 2:5139-5144.

6. Khalmatov Kh.Kh. Plants of Uzbekistan with diuretic action. Medicine, Tashkent, 1979.- P.126

7. Chervyakovsky, E. M. The role of flavonoids in biological reactions with electron transfer / E. M. Chervyakovsky, V. P. Kurchenko, V. A. Kostyuk // Proceedings of the Belarusian State University. Series: Physiological, biochemical and molecular bases of functioning of biosystems. - 2009. - V. 4. - No. 1. - S. 9-26.

Table 1
Diuretic activity of cuff juice Group
animals Dose mg/kg Urine volume, ml Diuretic index 1 hour 3 hours 5 o'clock Sum Intact 20 0.7±0.4 1.2±0.6 0.1±0.1 2.0±0.2 Experienced: Infusion of the cuff ordinary 20 0.1±0.1
* ^{1, 2} 1.4±0.1
* ² 1.6±0.2
* ¹ 3.1±0.1
* ^{1, 2} 1.6 Biostimulated cuff juice 20 1.5±0.5 ^*3 1.1±0.4
^*2 3.1±0.5 ^{*1, 2, 3} 5.6±0.6
^*13 2.8 Comparator infusion of wild strawberry 20 1.6±0.6 2.5±0.6
* ¹ 1.2±0.4
* ¹ 5.2±0.6
* ¹ 2.6

Note: p <0.05 * ¹ - compared with intact animals; * ² - in comparison with the reference drug, wild strawberry infusion; * ³ - compared with the infusion of common cuff.

table 2
Influence of juices from the grass of the common cuff on the concentration
sodium (mmol/l) and potassium (mmol/l) in urine Group
animals Dose mg/kg Concentration of elements, mmol/l for 5 hours. Saluretic index Na\K ratio Na K Na K Intact 182±14 247±10 1 1 0.7 Experienced: Infusion of the cuff ordinary 20 100±10
* ¹ 295±6
* ^{1, 2} 0.55 1.19 0.3 Biostimulated cuff juice 20 81±12
* ¹ 251±16
* ³ 0.44 1.01 0.3 Comparator infusion of wild strawberry 20 119±11
* ¹ 198±13
* ¹ 0.7 0.8 0.6

Note: p <0.05 * ¹ - compared with intact animals; * ² - in comparison with the reference drug, wild strawberry infusion; * ³ - compared with the infusion of common cuff.

Claims (1)

A method for obtaining a diuretic agent, which consists in the biogenic stimulation of fresh grass of the common cuff, collected in the phase of flowering and the beginning of fruiting, in darkness and cold for 7 days in a sealed package at a temperature of +4°C, followed by grinding to a particle size of 3 -5 mm, squeezing, preserving with 95% ethyl alcohol in the ratio juice : alcohol - 10:2, keeping the juice at a temperature of +4°C for 10 days, filtering.