RU2196596C2 - Virucidal, bactericidal, fungicide agent - Google Patents

Abstract

FIELD: medicine, pharmacology. SUBSTANCE: medicinal agent presents para-hydroxyacetophenone obtained from wood greens of coniferous strains by successive extraction with benzine, alkali aqueous solution and hydrocarbon extractant. Agent can be used as preparation exhibiting virucidal, bactericidal and fungicide effect for treatment of sicknesses caused by pathogenic viruses, fungi and microorganisms. EFFECT: expanded assortment of preparations of vegetable origin, enhanced virucidal, bactericidal and fungicide properties. 2 tbl

Description

 The invention relates to the field of medicine, pharmacology and can be used as an agent with virucidal, bactericidal and fungicidal action for the treatment of diseases caused by pathogenic viruses, fungi (yeast) and microorganisms.

 A number of herbal medicines are known (for example, from pine, spruce needles), which are used in the form of tinctures and oils to treat diseases such as tonsillitis, sinusitis, chronic pneumonia, etc. (A.F. Gammerman and others. Medicinal plants M.: Higher School, 1990, p. 94-100).

 It is known to use fir water, which is a waste in the production of fir oil, as a substance with anti-inflammatory, biostimulating and general strengthening effect (SU 1805966, A 61 K 35/78, 1986).

 Known agent with anti-inflammatory, antibacterial and wound healing activity (RU 2054945, A 61 K 35/78, 1991). It contains a capsular extract of Siberian fir 19.0-20.0, monoterpene compounds of essential oils 0.02-0.20 and vegetable oil - the rest.

 It is known to use the fraction of labdanic alcohols, a product of the processing of coniferous herbs, as a virucidal, bactericidal, fungicidal agent (RU 2138279, A 61 K 35/78, 1997).

 The problem to which the present invention is directed is to expand the arsenal of herbal preparations with more active virucidal, bactericidal, fungicidal properties.

 This problem is solved in that as a virucidal, bactericidal, fungicidal agent, steam-hydroxyacetophenone obtained from the processing of coniferous wood products is used.

 The method of obtaining steam hydroxyacetophenone is as follows. Shredded coniferous wood is extracted with gasoline. The resulting residue is freed from gasoline and subjected to extraction with an aqueous solution of alkali.

 Then the green wood is separated from the aqueous alkaline solution and the aqueous alkaline solution is acidified to pH 6-7 and extracted with a hydrocarbon extractant. Then the hydrocarbon extractant is distilled off and a residue is obtained which contains 60-70% purity hydroxyacetophenone.

 Studies have been conducted of the virucidal, bactericidal and fungicidal properties of parahydroxyacetophenone.

The following substances were taken for study and comparison:
1. Chlorophyllin sodium.

 2. Chlorophyllocarotene coniferous paste.

 3. Concentrate provitamin coniferous.

 4. Steam-hydroxyacetophenone isolated from European spruce needles.

 To study the antimicrobial activity of the obtained substances, we used the culture of Candida albicans strain 13 from the VKPG museum (Russian collection of pathogenic fungi), Staphylococcus aureus ATCC 6538 r, Escherichia coli ATCC 25922, Pseudomonas aureginosa from the GISK Museum named after Tarasovich.

 Candida albicans is a yeast-like fungus, the most common causative agent of candidiasis of various localization. Candidiasis is an opportunistic infection that occurs against the background of "risk factors" (the use of broad-spectrum antibiotics, corticosteroids, cytostatics, immunosuppressants for HIV infection, etc.).

 Staphylococcus aureus is a gram-positive microorganism, a frequent saprophyte on the skin and mucous membranes. The causative agent of inflammatory diseases of various localization.

 Escherichia coli is a gram-negative microorganism. It is found in the gastrointestinal tract in humans and animals. A common cause of cystitis, pyelonephritis, cholecystitis, septacemia, wound infections. Some strains are the cause of severe diarrhea in children.

 Pseudomonas aureginosa gram-negative microorganism. It is found in feces, on skin, in water, in air, in soil. Common causative agent of latent infections. Sometimes it is the cause of meningitis, respiratory, urinary, intestinal infections in children. Causes severe septicemia in immunocompromised patients.

 The experimental technique was as follows.

 The antimicrobial activity of substances 1-4 was determined by serial dilution. A suspension of yeast-like fungus was prepared on an isotonic solution of sodium chloride from 2-day-old cultures of Candida albicans with a density of 10 units. according to the bacterial standard (109 tank of cells per ml), which corresponds to 107 yeast-like organisms.

 Antibacterial activity was tested on a suspension of bacterial daily cultures with a density of 5 units. according to the bacterial standard.

Weighed portions of the test substance were dissolved in dimethyl sulfoxide at the rate of 40 mg / ml (maximum dissolution of a number of substances). For better solubility, many substances had to be heated to 80 ^o C.

 A number of tubes were prepared, in the first of which 1.8 ml of Saburo medium (for yeast-like mushrooms) or meat-peptide broth (for bacteria) was poured; in all subsequent - 1 ml of medium From a test tube with the initial solution of the drug, 0.2 ml was added to the first test tube of the row, from this test tube, after stirring, 1 ml was transferred to the next and so on until the end of the row. Thus, a series of dilutions in decreasing concentrations is obtained, a multiple of two (40-20-10-5, etc. mg / ml). Then, 0.1 ml of a suspension of the microorganism was added to each tube of the test series.

In each titration, there were three control tubes: culture control (test tube without test drug), drug control (test tube without culture). All control and working tubes were incubated for 2 days at 37 ^o C for studying yeast-like fungi and 1 day at 37 ^o C for studying the sensitivity of gram-positive and gram-negative bacteria.

 The minimum inhibitory concentration (MIC) of the test substance was considered dilution in the last tube, where there was no visible growth of the culture. To determine the minimum microbicidal concentration (MCC) from all test tubes where there was no visible fungal growth, 0.1 ml of the content was sown on Petri dishes with solid nutrient medium (Saburo agar for yeast-like and meat-peptone broth for bacteria). The lack of growth indicates the microbicidal effect of the drug in this dilution.

 The research results are given in table. 1-2.

 Of all the investigated substances, substance 4 (parahydroxyacetophenone from European spruce needles) should be distinguished as the substance with the highest fungicidal and bacterial activity. This substance was well soluble in DMSO, and therefore it was possible to easily analyze the ratio between MIC and MCC (4 MIC / MCC for Candida albicans - 0.5; for Staphylococcus aureus - 0.06; for E. coli - 0.5). The MIC / MCC ratios of substance 4 are closest to unity, i.e., this substance not only stops the growth of these microorganisms, but also kills them in close concentration.

 The activity of substance 4 is high both against yeast-like fungi and against gram-positive and gram-negative bacteria. This substance was also most active for other yeast-like and filamentous fungi, which are of great importance in medical and environmental terms.

 It should be noted that the investigated substance is significantly less active compared to modern antibiotics and synthetic drugs, where sensitivity is measured in μg / ml. However, given the natural origin and less toxicity, less skin irritating effect, this substance can become the basis for the development of cosmetics and detergents.

 Virucidal activity was evaluated by reducing the infectious titer of the virus with direct contact of influenza virus A / Hong Kong 2/68 (H3 2) and influenza B / Hong Kong-1/68 / C n-hydroxyacetophenone compared with the control. Chlorophyllocarotene paste was used as a reference preparation. The results are shown in table 2.

After 30 minutes of contact, para-hydroxyacetophenone inactivated 10 ³ infectious units of influenza A virus and 10 ² units of virus B. Chlorophyllocarotin paste at a concentration of 30 mg / ml reduced the infectious activity of influenza A and B viruses after 2 minutes by 100,000 times , and after 5 minutes the infectious virus in the solution was completely absent.

 The invention will find application in medicine as an agent having a virucidal, bactericidal and fungicidal effect in the treatment of diseases caused by pathogenic viruses, fungi (yeast) and microorganisms.

Claims (1)

 A drug with virucidal, bactericidal and fungicidal activity, which is parahydroxyacetophenone obtained from coniferous wood greens by sequential extraction with gasoline, an aqueous alkali solution and a hydrocarbon extractant.

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