WO2018093343A1

WO2018093343A1 - Active biological substance and a preparation based thereon

Info

Publication number: WO2018093343A1
Application number: PCT/UA2017/000109
Authority: WO
Inventors: Вадим Александрович САФОНОВ; Елена Юрьевна ГОЛИК; Николай Андреевич КОМИСАРЕНКО; Андрей Николаевич КОМИСАРЕНКО; Дарья Валентиновна НИКОЛОВА; Дмитрий Иванович ИСАЕВ; Дмитрий Михайлович КОЖУШКО
Original assignee: Вадим Александрович САФОНОВ
Priority date: 2016-11-15
Filing date: 2017-11-06
Publication date: 2018-05-24
Also published as: RU2647460C1

Abstract

The claimed invention relates to an active biological or pharmaceutical substance which has antibacterial activity and has a bactericidal, bacteriostatic and antifungal effect. According to the claimed invention, an active biological substance is characterized by the presence of: eucalyptus extract, benzalkonium chloride, and essential oils of eucalyptus, mint, fir, pine, lavender, tea tree, rosemary, coriander, rose, and sage in the following ratio of components in mass percent: eucalyptus extract 18.86-55.63, benzalkonium chloride 18.86-44.47, eucalyptus essential oil 3.70-22.24, mint essential oil 3.70-22.24, fir essential oil 0.1-4.42, pine essential oil 0.1-4.42, lavender essential oil 0.1-4.42, tea tree essential oil 0.1-4.42, rosemary essential oil 0.1-4.42, coriander essential oil 0.1-4.42, rose essential oil 1.34-2.24, and sage essential oil 1.34-2.24. The technical result of the invention is the production of an active biological substance and a topical preparation based thereon that have antibacterial activity and exert a bactericidal, bacteriostatic and antifungal effect.

Description

ACTIVE BIOLOGICAL SUBSTANCE AND DRUG ON ITS BASIS

5 Scope of the invention

The claimed invention relates to an active biological or pharmaceutical substance, which has antibacterial activity, has bactericidal, bacteriostatic and antifungal effects,

The invention also relates to a topical preparation obtained on the basis of the above active biological substance.

The invention can be used for the prevention and treatment of infectious and inflammatory diseases.

Also, the invention can be used to deodorize 15 oral cavities.

The invention can also be used to inhibit the growth activity of the strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633 and Candida albicans ATCC 653/885.

20 The claimed invention can be used in the chemical, pharmaceutical, cosmetic and food industries.

Prior art

An increase in the number of infections caused by various kinds of microorganisms, 25 which are resistant to many antimicrobial agents, requires the need to expand the arsenal of topical drugs that have antimicrobial effects, which are synthetic and semi-synthetic antibiotics, to which most microorganisms quickly develop resistance. In addition, these antimicrobial agents have a toxic effect on organs and systems of the human and animal body, contribute to the development of dysbiosis, hepatitis, and the like. The majority of local antimicrobials contain antiseptics of chemical origin, but they are xenobiotics for the body and therefore, against the background of infectious intoxication, cause an additional load of its metabolizing and eliminated systems, therefore, it is a possible cause of adverse reactions, especially in children, pregnant women, women during lactation and patients with chronic diseases. In this regard, herbal preparations are becoming more common, which are topical preparations that include herbal components (eucalyptus, mint, sage and others). As a result, herbal preparations contain complexes of biologically active substances that are close to the metabolites of the body and are biologically compatible with them. Another advantage of topical preparations of phytopreparations is their normalizing effect on altered body functions in the absence of influence on other indicators. These properties, as well as the use of low doses of plant components, can virtually eliminate toxic and side effects on the human body during their use.

The well-known drug “Chlorophyllipt” (Clorophylliptum), which is obtained from the leaves of ball eucalyptus (Eucalyptus globulus L) or rod eucalyptus (Eucaliptus viminalis. L) by extraction with 95% ethanol, which has antibacterial activity against staphylococci, streptococcus and A.S. Ns SU1123706). In practice, 0.25%, 1% alcohol and 2% oil solutions of Chlorophyllipt are used. The main biologically active substances of the drug are lipophilic substances: a mixture of euglobals (phenol aldehydes), terpenoids and chlorophylls (Kurkin VA, Pharmacognosy, Samara, LLC Etching, 2004, p. 327-333).

A solution of “Chlorophyllipt” (see A.S. see Nfi SU 801341) is obtained by extraction of the leaves of the eucalyptus rod (Eucaliptus viminalis. L) with 96% ethanol and subsequent processing of the concentrated extract with a divalent copper salt, namely 4% copper sulfate solution subsequent extraction of the target product is the amount of hydrophobic components. Extraction with benzene is carried out three times with stirring each time for 15 minutes and settling for 5-6 hours. The settled benzene extract is washed 3 times with purified water and evaporated in vacuo at a temperature of 40 ° C, and then dissolved in alcohol or in oil.

Other methods for producing “Chlorophyllipt” and preparations made on its basis are also known, see NSN ^Q SLM 123706, RU2320360, RU2416423, UA39463, UA79383 and UA92308.

The disadvantages of “Chlorophyllipt” and preparations made on its basis is that its antimicrobial effect is directed mainly to bacteria of the genus Staphylococcus and cannot be used against fungi. For example, the Nsl table shows the growth retardation diameter of 1% Chlorophyllipt extract.

Table Νδ1

The essence of the invention

The objective of the invention is the creation of an active biological substance that has antibacterial activity, has bactericidal, bacteriostatic and antifungal effects.

Another objective of the invention is the creation of a topical preparation, which is obtained on the basis of an active biological substance that has antibacterial activity, has bactericidal, bacteriostatic and antifungal effects. Another object of the invention is to suppress the growth activity of the strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633 and Candida aibicans ATCC 653/885.

Another objective of the invention is to increase the effectiveness of the spectrum of action for the prevention and treatment of infectious and inflammatory diseases.

Another objective of the invention is to increase the effectiveness of the spectrum of action during deodorization of the oral cavity.

It is also an object of the invention to reduce the addiction (resistance) of microorganisms to an active biological substance and, based on it, a locally produced preparation.

Another objective of the invention is to reduce the toxicity of exposure to the body during its use.

It is also an object of the invention to expand the arsenal of means for suppressing the growth activity of strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633 and Candida aibicans ATCC 653/885.

Other objectives and advantages of the invention will be disclosed as the description and tables.

The above objectives are achieved as a result of the creation of an active biological substance, which is characterized by the presence of an extract of eucalyptus, benzalkonium chloride and essential oils of eucalyptus, mint, fir, pine, lavender, tea tree, rosemary, coriander, rose and wormwood with the following ratio of components, May. %:

eucalyptus extract 18.86-55.63

benzalkonium chloride 18.86-44.47

Eucalyptus essential oil 3.70-22.24

peppermint essential oil 3.70-22.24

fir essential oil 0, 1-4,42 pine essential oil 0,1-4,42 lavender essential oil 0.1-4.42

tea tree essential oil 0.1-4.42

rosemary essential oil 0.1-4.42 coriander essential oil 0.1-4.42 rose essential oil 1, 34-2.24

wormwood essential oil 1, 34-2.24

Also according to the invention, the above active biological substance additionally contains 1-15% polyvinylpyrrolidone.

Also according to the invention, the above active biological substance additionally contains 10-50% polysorbate 80.

Also according to the invention, the preferred composition of the aforementioned active biological substance has, in the following ratio of components, May. %:

eucalyptus extract 10.3

benzalkonium chloride 10.3

essential eucalyptus 5.15

oil

peppermint essential oil 5.15

essential oil of fir 1, 03

pine essential oil 1, 03

lavender essential oil 1, 03

tea essential oil 1, 03

tree

rosemary essential oil 1, 03

coriander essential oil 1, 03

rose essential oil 0.51

wormwood essential oil 0.51

polysorbate 80 52.41

polyvinylpyrrolidone 10.3

Not having the intention to associate themselves with any particular theory, the inventors believe that the combination of active components an extract of eucalyptus, benzalkonium chloride and a complex of essential oils in an active biological substance, provides a synergistic pharmacological action, as a result of which the use of the obtained active biological substance leads to inhibition of the growth activity of the strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCConas aerosas P36 4636, Proteus vulgaris ATCCosaas3636, ATCC 27853, Basillus subtilis ATCC 6633, and Candida albicans ATCC 653/885. The resulting active biological substance has antibacterial activity, has bactericidal, bacteriostatic and antifungal effects.

The use of polyvinylpyrrolidone in an active biological substance leads to the prolongation of the antimicrobial and antifungal action, which consists in the fact that the presence of polyvinylpyrrolidone creates a film on the surface of the organ that is being processed, and this leads to improved adhesion, which leads to the prolongation of the antimicrobial and antifungal action .

The use of polysorbate 80 in the active biological substance leads to the fact that the stability of the solution is increased.

A topical preparation characterized by the fact that it contains an active biological substance according to the invention, glycerin and purified water in the following ratio, May. %

active biological substance 0.5-2

glycerin 0.5-5.0 purified water the rest

A package of a topical preparation according to the invention is a topical preparation that is placed in a bottle with a metering device.

Nfi example 1

To prepare an active biological substance, the active components are mixed: eucalyptus extract, benzalkonium chloride and essential oils of eucalyptus, mint, fir, pine, lavender, tea tree, rosemary, coriander, rose and wormwood, and with the addition of additional components of polyvinylpyrrolidone and polysorbate 80.

Table Ne2 shows the three obtained samples of active biological substance that suppress the growth activity of the strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633 and Candida albicans ATCC 6633.

Peppermint oil was used as peppermint oil. Table Νδ 2

Composition Samples of biologically active

substances, may. %

1 2 3 eucalyptus extract 5.3 17.3 10.3 benzalkonium chloride 5.3 10.3 10.3 essential eucalyptus oil 5.15 1.15 5.15 peppermint essential oil 5.15 1, 15 5.15 essential fir oil 1.03 0.03 1.03 pine essential oil 1.03 0.03 1.03 lavender essential oil 1.03 0.03 1.03 tea essential oil 1.03 0.03 1.03 tree

rosemary essential oil 1.03 0.03 1, 03 coriander essential oil 1.03 0.03 1.03 rose essential oil 0.51 0.51 0.51 wormwood essential oil 0.51 0.51 0.51 polysorbate 80 61, 60 58.60 51, 60 polyvinylpyrrolidone 10.3 10.3 10.3 The obtained samples of the active biological substance have a color from green to greenish-brown, an shelf life of 36 months, and have an antibacterial and antifungal effect. Example Ns 2

Based on the obtained sample of Ns3 of an active biological substance, see Example Ns1, a topical preparation was prepared. For this, purified water was mixed with glycerin followed by the addition of an active biological substance (sample Ns3), as a result of which three samples of a topical preparation were obtained, see table Νδ3.

Table Ns 3

After that, the antibacterial and antifungal activity of the obtained sample Ν ^β 3 “Chlorophyllipt Asset Plus” was determined. To compare the results, we took a control drug - Chlorophyllipt spray, which was manufactured by GNTsLS (State Scientific Center for Medicines).

A microbial suspension of strains (microorganisms) was prepared, which was carried out using a Densi- La-Meter device (manufactured by PUVA-Lachema, Czech Republic; wavelength 540 nm). Microbial suspension of the strains was carried out according to the instructions that are attached to the device, and the newsletter on innovations in health system "Standardization of the preparation of microbial suspensions", Kiev. The culture of the strains was synchronized using a low temperature (4 ° C). Microbial loads were 10 ⁷ microbial cells per 1 ml of medium and were established according to McFarland standard 5. An 18-24 hour culture of microorganism strains was taken into the work. For research, Muller-Hinton agar was used (Production - India “HIMediaLaboratorles Pvt. Ltd lndia”, the shelf life of the medium is until XI.2016). Saburo agar was used for Candidaalbicans (Production - India, “HIMedial_aboratorles Pvt. Ltd lndia” expiration date wednesday until X1.2016).

The method of diffusion of the drug into agar was carried out by "wells". The determination of the activity of antibacterial drugs was carried out on two layers of a dense nutrient medium spilled in Petri dishes. In the lower layer used a "hungry" un seeded medium (agar-agar, water, salt).

15 The bottom layer is a substrate of 10 ml of “hungry agar”, on which 3-6 thin-walled stainless steel cylinders with a diameter of 8 mm and a height of 10 mm are installed horizontally. Around the cylinders is poured the upper layer, consisting of a nutrient agar medium, molten and cooled to 40 ° C, into which

20 corresponding standard daily culture of test strains. In advance, the top layer was well mixed until a homogeneous mass was formed. After solidification, the cylinders were pulled out with sterile tweezers, and test preparations (Chlorophyllipt, manufactured by GNTSLS (patent RU 39463) and sample Νδ3 "Chlorophyllipt asset were placed in the wells formed

25 plus "), taking into account its volume (0.3 ml).

The volume of medium for the upper layer ranged from 14 to 16 ml. The cups were dried for 30-40 minutes at room temperature and placed in a thermostat for 18-24 hours.

When evaluating the antimicrobial and antifungal properties of the drugs, the following criteria were used:

- the absence of growth retardation zones of strains around the well, as well as a delay zone of up to 10 mm indicates that the strains are not sensitive to the local preparation of the studied drug introduced into the well;

- growth retardation zones with a diameter of 10-15 mm indicate a low sensitivity of the strains to a topical preparation;

- growth retardation zones with a diameter of 15-25 mm are regarded as an indicator of the sensitivity of the strains to a topical preparation;

- growth inhibition zones, the diameter of which exceeds 25 mm, indicates a high sensitivity of the strains to the drug local action.

To obtain reliable data to draw a conclusion regarding the effectiveness of the drugs, three independent trials were performed (number of repetitions of studies). Test data of antibacterial and antifungal activity are set out in table Νδ4.

Table 4

The data in table Ns4 indicate that, in contrast to the known preparation, the invention actively suppresses the growth of strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Basillus subtilis ATCC 6633, and also provides static suppression of the growth of the fungus strain Candida albicans ATCC 653/885 , that is, the resulting topical preparation obtained on the basis of biologically active substance has antibacterial activity, has bactericidal, bacteriostatic and antifungal effects.

It should be further noted that the use of 5 mint and eucalyptus oils provides deodorization of the oral cavity.

From the data in table N ° 4 it is seen that the concentration of the active biological substance of 0.97% in a topical preparation indicates the high efficiency of the active biological substance and that its use does not lead to irritation, which causes a high concentration of ethyl alcohol in the known preparation " Chlorophyllipt, "and also indicates that a topical preparation can be effectively used against catarrh and for the prevention and treatment of infectious and inflammatory diseases of the upper respiratory tract ial tract.

15 The resulting topical preparation can be used in the form of a solution, spray and aerosol.

A package of a topical preparation is the resulting topical preparation, which is placed in a bottle with a dosing device, with the help of which the topical preparation

20 is applied to the oral cavity.

Title to the invention

We, the inventors, the obtained active biological substance, which has antibacterial activity, has 25 bactericidal, bacteriostatic and antifungal effects called: lat. "Clorophylliptum active plus", Ukrainian Χηοροφίπίπτ Asset Plus, Rus. "Chlorophyllipt asset plus."

It should be clear that this description provides the necessary and sufficient data for a clear and complete understanding of the invention.

Information that is obvious to specialists in this field of technology and which would not contribute to a better understanding of the invention was not given in this description. For example, it is clear that an active biological substance can be used as an antiseptic for scratches and burns.

Technical result

The technical result of the invention is the production of an active biological substance and a local preparation based on it, which have antibacterial activity, have bactericidal, bacteriostatic and antifungal effects.

Claims

CLAIM

1. An active biological substance with antibacterial activity and antifungal activity, characterized by the presence of: eucalyptus extract, benzalkonium chloride and 5 essential oils of eucalyptus, peppermint, fir, pine, lavender, tea tree, rosemary, coriander, rose and wormwood in the following ratio of components, May. %:

eucalyptus extract 18.86-55.63

benzalkonium chloride 18.86-44.47

Eucalyptus essential oil 3.70-22.24

peppermint essential oil 3.70-22.24

essential oil of fir 0.1-4.42

pine essential oil 0.1-4.42

lavender essential oil 0.1-4.42

tea tree essential oil 0.1-4.42

rosemary essential oil 0.1-4.42

coriander essential oil 0.1-4.42

rose essential oil 1, 34-2.24

wormwood essential oil 1.34-2.24

2. The substance according to claim 1, which additionally contains 1-15% polyvinylpyrrolidone.

s 3. The substance according to any one of the above paragraphs 1 or 2, which additionally contains 10-50% polysorbate 80.

4. The substance according to claim 3, which has a predominant composition in the following ratio of components, May. %:

eucalyptus extract 10.3 benzalkonium chloride 10.3

essential eucalyptus 5.15

oil

peppermint essential oil 5.15

essential oil of fir 1, 03

pine essential oil 1, 03 lavender essential oil 1, 03

tea essential oil 1, 03

tree

rosemary essential oil 1, 03

coriander essential oil 1, 03

rose essential oil 0.51

wormwood essential oil 0.51

polysorbate 80 52.41

polyvinylpyrrolidone 10.3

5. The substance according to claim 1, characterized in that it is used to inhibit the growth activity of the strains: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633 and Candida

5 albicans ATCC 653/885.

6. A topical preparation with antibacterial activity and antifungal activity, characterized in that it contains an active biological substance, characterized according to any one of the above items 1-5, glycerin and water purified in the following ratio of components, May. %:

active biological substance 0.5-2 glycerol 0.5-5.0

purified water

7. The drug according to claim 6, which is used to deodorize the oral cavity.

8. The drug according to claim 6, which is used for the prevention and treatment of infectious and inflammatory diseases of the upper respiratory

15 ways.

9. The drug according to claim 6, placed in a bottle with a metering device.

twenty