RU2175247C1 - Antiseptic composition and method of its preparing - Google Patents

Abstract

FIELD: medicine, sanitary hygiene. SUBSTANCE: invention proposes a composition that can be used for humans protection from pathogenic microorganisms. Said composition consists of the following components, wt.%: clathrate carbamidedidecyldi-methylammonium bromide, 10.0-30.0; organic stabilizing agent, 5.0-15.0; accessory substances, 0.01-2.0; water, up to 100%. Composition can contain additionally chelating agent, anticorrosive substance, antifoaming agent, antifreeze. Method of preparing an antiseptic composition involves mixing clathrate carbamidedidecyldimethylammonium bromide with different components being at first clathrate is dissolved in water thoroughly using shaking or rotor device for 1-2 h, settled for 2-10 h followed by by-step addition of stabilizing agent and different components and mixture is stirred thoroughly at each stage for 0.5-2.0 h. Composition shows ability to inactivate the broad spectrum of pathogenic microorganisms including tuberculosis mycobacteria and hepatitis B virus. EFFECT: enhanced effectiveness of composition. 16 cl, 4 tbl, 3 ex

Description

 The invention relates to the field of medicine and sanitary hygiene, in particular to the creation of a composition based on a derivative of a Quaternary ammonium compound having antiseptic and disinfecting effects.

 Various ammonium derivatives are widely used as antiseptic and disinfectants. In particular, preparations are known, such as an antimicrobial preparation based on quaternary ammonium and pyridinium compounds with a halogen (SU 557793, 1977), a virucidal preparation containing dimethylacetylbenzylammonium chloride (SU 1796185, 1979), a fungicidal preparation based on a derivative benzylammonium (SU 1796185, 1990), the same drug has antiseptic and disinfecting effects (WO 93/00892, 1993).

 The antimicrobial activity of quaternary ammonium derivatives is also described in international applications: WO 83/01002 (03.31.83), WO 90/04918 (05.17.90) and WO 98/56366 (17.12.98).

 However, the above funds have such disadvantages as a narrow focus, some of them contain chlorine, which is toxic to humans and warm-blooded animals.

 Closest to the present invention are developments that describe the use of such a quaternary ammonium compound as a water-soluble clathrate of didecyldimethylammonium urea bromide (Israeli patent 58586, 31.08.82, WO 97/27880, 07.08.97, RU 2095086, 10.11.97). The named water-soluble clathrate has a fairly wide range of antimicrobial and antiviral effects, does not contain chlorine and is currently used as a disinfectant.

The empirical formula of the clathrate compound of urea-didecyldimethylammonium bromide (CDDAB) is C ₃₈ H ₁₁₂ N ₃₃ O ₁₆ Br.

Молекулярная масса его - 1367,5.Structural formula:

Its molecular weight is 1367.5.

Known disinfectant composition containing the above clathrate (KDAB) and other ingredients (WO 97/27880, 08/07/97), which is a mixture of components in the following proportions, wt.%:
Clathrate KDAB - 81-22
Lower alcohol (mainly ethyl) - 18-22
Water - 56-64
From the description of the aforementioned international application it follows that this composition has bactericidal, as well as virucidal and fungicidal activity.

 The named disinfectant composition is prepared as follows. 400 ml of distilled water is placed in a container. 200 mg of 96% ethanol are added, 200 g of KDDAB clathrate are added, and 200 ml of distilled water are added. After this, the mixture is stirred and allowed to stand for 2 hours. The resulting solution was filtered.

 However, our own studies have shown that the drug obtained in this way is not active enough against a currently relevant virus such as hepatitis B virus, does not destroy poliomyelitis virus, and, moreover, is not capable of complete (100%) disinfection of medical equipment contaminated with blood and other substrates of biological origin.

The technical result of the present invention is to remedy these disadvantages. The problem is solved by developing a new antiseptic composition based on an aqueous solution of carbamide didecyldimethylammonium bromide clathrate (KDAB), which additionally contains an organic stabilizer and auxiliary substances in the following ratio of components, wt.%:
Clathrate KDAB - 10.0-30.0
Organic Stabilizer - 5.0-15.0
Excipients - 0.01-2.0
Water - Up to 100.0
This composition may further comprise complexon in an amount of 0.1-0.2 wt.%.

 The expansion of the spectrum and enhancing the action of the composition is directly related to the improvement of the technology for its preparation, therefore another aspect of the invention is to provide a method for preparing an antiseptic composition. It lies in the fact that in the method of preparing an antiseptic composition by mixing the KDDAB clathrate with various components, a distinctive feature is that at first the KDDAB clathrate is thoroughly dissolved in water using a shaking or rotary device (for example, a mixer) for 1-2 hours, they stand for 2-10 hours, then a stabilizer (for example, alcohol or glycol) and additional components are added in stages with thorough mixing at each stage for 0.5-2.0 hours, they stand for 2-3 hours, filtered, and poured into bone and sealed.

To create a composition as an organic stabilizer can be used a number of components. Alcohols containing carbon atoms from 1 to 3 (C ₁ -C ₃ ), acetone, glycol or its derivatives.

 The claimed composition may additionally contain complexon.

 As a complexon, this composition may contain ethylenediaminetetraacetic acid (EDTA) or its derivatives. Compounds of the complexon type included in the present composition play a significant role in promoting the solubilization of biological substrates present on disinfected objects: blood clots or lymph on surgical instruments, the remains of the contents of the examined body cavities on individual parts of endoscopes, catheters, rubber and plastic tubes, various surgical and diagnostic tools.

 Substances such as EDTA in this case act as emulsifiers, which ultimately increase the bactericidal and virucidal effect to 100%.

 The organic stabilizer introduced into the composition improves the stability of the drug in aqueous solution, allowing you to save the composition without loss of activity up to 3-5 years.

The most optimal and studied variant of the claimed composition is a variant that contains KDAB clathrate, isopropyl alcohol as an organic stabilizer, perfume as an auxiliary substance, as well as water in the following ratio of components, wt.%:
Clathrate KDAB - 20.0 ± 1.0
Isopropyl alcohol - 10.0 ± 2.0
Fragrance - 0,01
Water - Up to 100.0
This composition, conventionally designated as "Dezavik", has a number of positive qualities and can be used as an antiseptic and disinfectant. "Desavik" has a wide range: very high (up to 100%) bactericidal and virucidal activity in relation to:
gram-positive and gram-negative bacteria, including pathogens of salmonellosis, typhoid fever, dysentery, diphtheria, scarlet fever, sexually transmitted diseases, as well as tuberculosis (tested on mycobacteria B ₅ );
especially dangerous infections (cholera, anthrax, plague);
influenza viruses, parainfluenza, herpes, HIV infection, hepatitis A, B and C (100% destruction);
anaerobic infection (gangrene, tetanus, botulism);
fungi of the genus Candida and dermatophytes.

 The composition "Dezavik" has both a washing and deodorizing effect, in addition, it has anti-corrosion properties, which are enhanced when additional anti-corrosion agents, for example, nitrates or phosphates of alkali metals, are added to the composition.

In addition to the well-known use of products containing a water-alcohol solution of KDDAB clathrate for disinfecting surfaces in the premises of communal facilities and medical institutions, as well as disinfecting equipment, implements and containers in the dairy and meat industry, the present invention is directed to solving health problems during antiseptic processing of the following objects:
medical devices, including surgical and diagnostic tools, flexible and rigid endoscopes, parts of medical devices contaminated with biological substrates (blood, lymph, various exo- and endoviruses, etc.);
dental instruments and devices, including those heavily contaminated with biosubstrates;
gynecological and urological diagnostic and therapeutic devices and auxiliary equipment;
patient care items (including those contaminated with blood and other biosubstrates), dishes, linen, etc.

 The present composition may include a number of additional excipients, which allow more efficient use in various directions as a universal antiseptic and disinfectant.

The composition contains as auxiliary substances:
fragrance, for example, with the smell of lemon, apple, cherry, etc .;
anticorrosive substance, mainly sodium nitrate or disodium phosphate;
anti-foaming agent, for example amyl or octyl alcohol;
antifreezes - glycerin or ethylene glycol.

 The method for preparing the composition also provides for the additional introduction of components such as complexone, perfume, anti-corrosion agent, anti-foam agent, antifreeze, or a combination thereof.

When implementing the method, it should be borne in mind that C ₁ -C ₃ alcohol, acetone, glycol or its derivative can be added as a stabilizer, sodium nitrate or disodium phosphate as an anticorrosive substance, EDTA or its derivative as a complexon, as antifoam agent - amyl or octyl alcohol, as antifreeze - glycerin or ethylene glycol. An additional component is introduced in stages after the introduction of the stabilizer, each time thoroughly mixing the mixture using available devices, such as a shaking device or a rotary mixer.

For a better understanding of the invention, specific formulations of the compositions:
Example 1: Classical antiseptic composition for disinfection of premises and equipment, wt.%:
Clathrate KDAB - 15.0 ± 1.0
Isopropyl alcohol - 10.0 ± 1.0
Complexon III - 0.1
Fragrance with a smell of a lemon - 0,02
Water - Up to 100.0
The recipe of the composition described above is prepared as follows: first, the KDDAB clathrate is mixed with water using a rotary mixer for 1 hour, sedimented for 3 hours, then isopropyl alcohol is added, it is stirred for 30 minutes, it is left to stand for 2 hours, complexon III and perfume are added, again thoroughly mix for 1 hour, stand for 2 hours, filter, pour into containers and seal.

Example 2: Antiseptic composition for use in areas with predominantly low-temperature atmospheric conditions, wt.%:
Clathrate KDAB - 25.0 ± 1.0
Ethylene glycol - 12.0 ± 1.0
EDTA - 0.1
Octyl alcohol - 0.2
Fragrance - 0.04
Water - Up to 100.0
The above composition is prepared according to the technology similar to that described in example 1.

Example 3: Antiseptic composition with predominant use for: metal surfaces, containers and tools:
Cladrate KDAB - 14.0 ± 1.0
Propyl alcohol - 7.0 ± 1.0
Ethylenediaminetetraacetic acid trisodium salt - 0.2
Fragrance - 0.03
Disodium Phosphate - 0.6
Water - Up to 100
This composition is obtained using the methodology described in detail above and similarly described in example 1.

 All studies on antiseptic and disinfectant properties were carried out on the drug under the code name "Desavik", the composition of which is given above. For the named tool, tests were also conducted on its toxicity.

 The drug "Desavik" is currently the most studied and is the main representative of the claimed composition, including and summarizing the properties of the claimed composition.

 The study of the antiseptic and disinfectant properties of the agent was carried out in accordance with the “Methods of testing disinfectants to assess their safety and effectiveness”, Moscow, Ministry of Health of the Russian Federation, 1998, part 2. The strains resistant to disinfectants were used as test microorganisms: Staphylococcus aureus (strain 906), Escherichia coli (pcs. 1257), mycobacteria B5, candida albicans (pcs. 15), gypsum-type trichophytone poliomyelitis type 1 (vacc. pcs. Lsc 2ab).

 Medical devices were tested as test objects, including utensils, underwear, rubber tubes, micropipettes, forceps, tweezers, spatulas, burs, dental mirrors, pulp extractors, endoscopes, etc. (including those contaminated with biosubstrates).

 The results of the study of the antiseptic and disinfectant properties of the product "Dezavik" for infections of bacterial etiology (except for tuberculosis) are presented in table 1, viral etiology (hepatitis B, HIV infection) - in table 2, for tuberculosis - in table 3.

 As a result of the studies, it was found that various test surfaces of medical objects, as well as dishes without food residues, contaminated with Escherichia coli and Staphylococcus aureus, were disinfected with 0.25-0.5% drug solutions. Contaminated objects (dishes, some medical facilities, underwear) were disinfected with a 1.0 - 2.0% solution of the product.

 In relation to candida albicans and gypsum trichophytone, 2.0-4.0% solutions of the composition "Desavik" were effective (table 4).

 To study the virucidal effect against hepatitis B virus, the composition was diluted with water to a concentration of 2.0; 2.5 and 3.0%.

 After the objects were infected with the patient’s blood serum containing the hepatitis B virus surface antigen (HBsAG), the decontamination was performed by immersion in an excess of the drug solution. The exposure time with the antiseptic was 60 minutes at room temperature. After 60 minutes of processing, liquid samples were taken into which infected objects were immersed after washing them.

 An enzyme-linked immunosorbent assay for the detection of HBsAG was performed according to the instructions for use of the Hepanostika HBsAG Uni-Foriri II test system from Organon.

 The composition "Desavik" at a concentration of 2.5% for 60 min led to complete (100%) inactivation of HbsAG (see table. 2).

 We conducted a detailed study of the toxicity and various types of hazards of the claimed composition on the example of the composition "Dezavik".

 The objective of the research was to assess the toxicity and danger of the drug, including determining the average lethal dose for potentially dangerous routes (in the stomach, parenteral, i.e. intraperitoneal administration, and application to the skin), locally irritating properties of the composition in contact with intact skin integument (once and repeatedly) and with a single exposure to the mucous membranes of the eye.

 The inhalation hazard of the product was determined in two series - under conditions of saturating concentrations of volatile components and its working solution and maximum concentration (4%); determination of the sensitizing properties of the drug, as well as the development of a washing regime with an assessment of the safety of residual quantities after washing the components of the drug according to the resistance of red blood cells.

 In the experiment, small laboratory animals (white outbred mice and rats, as well as chinchilla rabbits) were used, which received a generally accepted diet containing all the necessary components for their normal life.

 Statistical processing of the obtained data was carried out using Student-Fisher. For statistically significant changes, the confidence level was taken at P <0.05.

The average lethal dose of the composition was determined on white outbred rats weighing 200-220 g in the range of doses of the drug from 1000 to 5000 mg / kg. The clinic of poisoning in animals was characterized initially by excitement, then impaired coordination of movement, increased breathing, the presence of a lateral position and their death. LD ₅₀ amounted to 2606 mg / kg (according to the classification of GOST 12.1.007-76, the product belongs to the 3rd class of moderately hazardous drugs).

When administered parenterally, LD _{50 was also} determined in the dose range of 200 to 800 mg / kg. LD ₅₀ when administered intraperitoneally for a disinfectant was 456 ± 42 mg / kg (according to the classification of K.K. Sidorov, the drug belongs to the 4th class of low toxicity).

 When applying the drug in a dose of 2500 mg / kg on intact skin, no death of animals was observed during the 14 days of observation (according to the classification GOST 12.1.007-76, the agent belongs to the 4th class of low-hazard drugs).

 Thus, the product in accordance with the classification of GOST 1.12.007-76, when introduced into the stomach, belongs to the 3rd class of moderately hazardous, when administered parenterally to the 4th class of low toxicity and when applied to the skin, it belongs to the 4th class of low-hazard drugs.

 The inhalation danger of the agent under conditions of exposure to saturating concentrations of volatile components showed that animals showed slight signs of irritation to the mucous membranes of the upper respiratory tract (scratching the muzzle with paws, sneezing, closing the eyes, increasing breathing). After the exposure was completed, after 5-10 minutes, the animals of the experimental group responded adequately to the animals of the control group in terms of appearance, behavior and responses. During the follow-up period (14 days), no animal deaths were recorded.

 Pairs of working solutions in concentrations up to 4% are not inhalation dangerous, since there are no signs of irritation to the mucous membranes of the nasopharynx and eyes, as well as changes in the functional state of the nervous system and respiratory rate.

 Thus, the volatile components of the product concentrate belong to the 3rd class of moderately hazardous compounds. According to the clinical signs of intoxication and objective data, the volatile components of working solutions up to 4.0% concentration do not have a general toxic and irritating effect on the mucous membranes of the upper respiratory tract and their vapors are inhalation-free and can be classified as class 4 low-hazard substances.

 The inhalation hazard of the agent with the maximum working concentration (4%) was carried out by wiping at a 10-fold overestimated flow rate in small-volume chambers, creating also static exposure conditions.

Thus, the acute action threshold of the “Dezavik” composition in the application mode by wiping is higher than 10 consumption rates. Based on the data obtained, it is possible to calculate the zone of acute toxic effects as the ratio of the value of the threshold of acute effects to the rate of consumption. This value amounted to more than 10 (Lim _ac / N _{flow rate} or> 10 N _{flow rate} / N _{flow rate} => 10), class 4 low hazard disinfectants. This value gives reason to recommend the use of the composition "Dezavik" in medical institutions in the presence of patients and patients, as well as the use in everyday life.

 The determination of the local irritating effect of the composition was determined on rabbits with a single and repeated exposure and working solutions in concentrations of 0.25; 2.0 and 4.0%. The product was applied to a clipped skin in the middle part of the spine. In this case, the left side served as a control, and the right side - for applying the drug. The skin reaction was evaluated in points on a scale of S.I.Suvorov.

 Observation of the skin condition of the rabbit showed that with a single contact with the skin, the test substance causes mild irritation in the form of skin hyperemia (2 points). With an increase in the number of applications, signs of irritation increased in the form of an increase in the severity of the initial signs, and subsequently the appearance of crusts (after 3 applications). Applications of the drug are discontinued. The healing of the affected skin took a long time (more than 15 days). Working concentrations of the agent up to 4.0% with a single exposure do not have a local irritant effect. With an increase in the duration of applications, a locally irritating effect was established upon repeated contact in the form of weak hyperemia (1 point) in a 2.0% working solution and more pronounced in a 4.0% solution. The first signs of irritation appeared on the 3-4th day of exposure (subsequent application of the solution was discontinued). On the affected area subsequently appeared dryness and small-scaled peeling.

 When 1-2 drops of the concentrate are introduced into the rabbit’s eye, a sharp painful reaction is observed, accompanied by sharp signs of irritation (hyperemia, blepharospasm, discharge from the eyes with the formation of further eyesight and loss of vision).

 The inflammatory reaction disappears by the end of 15-18 days.

 When 2 drops of the working solution of the agent in 0.25% concentration were introduced into the conjunctival sac of the eye, it caused slight irritation of the mucous membrane without damage to the conjunctiva. Changes disappeared after 2-3 days. Solutions in high concentrations caused an increase in irritation of the mucous membranes of the eyes. Based on the data obtained, we can conclude that the tool causes pronounced irritation of the mucous membranes of the eyes, and working solutions cause mild irritation.

 Thus, the concentrate has a pronounced local irritant effect (especially upon repeated contact with intact rabbit skin and eye mucosa). Working solutions of the composition also have a local irritant effect, but their severity decreases from 4.0% concentration to 0.5% concentration.

 To evaluate the sensitizing activity of the composition, a delayed-type hypersensitivity reaction (HRT) was used on white mice. For these purposes, Freund's complete adjuvant, manufactured by Hoechst (USA), was used. After preparing solutions based on Hanks and Freund's adjuvant, a sensitizing dose of 0.06 ml was administered and after 5 days a resolving agent based on Hanks solution in a volume of 0.04 ml was introduced. A sensitizing dose was administered at the base of the tail, and the resolving dose was injected into the pad of the left hind paw.

 The tool has a sensitizing activity. At the same time, the length of the foot in the control was 2.02 ± 0.026 mm versus 2.08 ± 0.026 mm in the experiment. The reaction index is 2.98%, which is at the level of an acceptable level, which is 5.0%.

 Testing the recall mode for medical devices was carried out on rubber with high sorption ability and low washability of the material according to the method “hemolytic activity of extracts of polymer materials and medical products based on them” using red blood cells. For this, the most severe disinfection and pre-sterilization treatment was taken (4% solution for 60 minutes), after which they were washed under running tap water for different times (1 and 3 minutes), followed by rinsing with distilled water for 30 s. Evaluation of the results of washability was carried out by the percentage of hemolysis of red blood cells (optical density).

Based on the data obtained in the study of the toxicity of the composition "Dezavik" we can draw the following conclusions:
according to the parameters of acute toxicity, the drug, when introduced into the stomach and applied to the skin, belongs to class III - IV moderately and slightly hazardous preparations in accordance with the classification GOST 12.1.007-76 (LD ₅₀ for rats 2606 mg / kg and more than 2500 mg / kg, respectively ); with intraperitoneal administration - to the 4th class of low toxicity agents according to the classification of K.K. Sidorov (LD ₅₀ for rats 456 ± 42 mg / kg);
vapors of the inhalation agent are dangerous in conditions of saturating concentrations and are characterized as moderately hazardous (hazard class 3), vapors of 2-4% working solution and lower hazardous (class 4 low hazard drugs);
with a single exposure, the native preparation irritates intact skin. With repeated applications, it has a pronounced local irritant effect. Working solutions in 2-4% concentration cause slight skin irritation upon repeated exposure;
upon contact with the mucous membranes, the drug causes their severe irritation, and working solutions at a concentration of 1% - 4.0% - mild irritation of the mucous membranes;
the drug does not have a sensitizing effect;
the following washing of residual quantities of the drug from medical devices depending on the material is recommended: glass and metal for 2 minutes, and rubber and plastic for 3 minutes.

 Thus, the claimed composition obtained using the modified preparation technology has low toxicity with a wider spectrum of antiseptic effects: antimicrobial (including tuberculosis mycobacteria), antiviral (including 100% destruction of hepatitis B virus and polio), highly effective action with antiseptic treatment complex medical devices contaminated with biosubstrates.

 The claimed composition, obtained using the modified cooking technology, can be successfully used in areas with different weather conditions (including high humidity, low temperature, etc.).

 It can be described as a universal antiseptic and disinfectant that can help solve the problem of epidemiological tension not only in hot spots, refugees, other similar emergency situations, but also throughout the country.

Claims (14)

1. An antiseptic composition containing an aqueous solution of carbamide didecyldimethylammonium bromide clathrate (KDAB), characterized in that it further contains an organic stabilizer and auxiliary substances in the following ratio of components, wt.%:
Clathrate KDAB - 10.0 - 30.0
Organic Stabilizer - 5.0 - 15.0
Excipients - 0.01 - 2.0
Water - Up to 100.0
2. The composition according to p. 1, characterized in that it further comprises complexon in an amount of 0.1 - 0.2 wt.%. 3. The composition according to claim 1, characterized in that it contains KDDAB clathrate, isopropyl alcohol as a stabilizer, perfume as an auxiliary substance, as well as water in the following ratio of components, wt. %:
Clathrate KDAB - 20.0 ± 1.0
Isopropyl alcohol - 10.0 ± 2.0
Fragrance - 0,01
Water - Up to 100
4. The composition according to PP.1 and 3, characterized in that as an organic stabilizer contains a substance selected from the group: alcohol with a carbon atom content of 1 to 3, acetone, glycol or its derivative.  5. The composition according to claim 2, characterized in that as complexone contains ethylenediaminetetraacetic acid or its derivative.  6. The composition according to claims 1 and 3, characterized in that as an auxiliary substance it contains fragrance, anticorrosive substance, antifoam agent, antifreeze, or a combination thereof.  7. The composition according to claim 6, characterized in that as the anti-corrosive substance contains sodium nitrate or disodium phosphate.  8. The composition according to PP.1 and 6, characterized in that as an auxiliary anti-foaming agent contains amyl or octyl alcohol.  9. The composition according to PP.1,3,6 and 8, characterized in that as an auxiliary substance contains glycerin or ethylene glycol.  10. A method of preparing an antiseptic composition by mixing carbamide didecyldimethyl ammonium bromide clathrate with various components, characterized in that at the beginning the clathrate is thoroughly dissolved in water using a shaking or rotary device for 1 to 2 hours, standing for 2 to 10 hours, then introduced in stages stabilizer and additional components, again thoroughly mix the mixture at each stage for 0.5 - 2.0 hours, stand for 2 - 3 hours, pour into containers and seal. 11. The method according to claim 10, characterized in that C ₁ -C ₃ alcohol, acetone, glycol or a derivative thereof are introduced as a stabilizer.  12. The method according to claim 10, characterized in that after the stabilizer, complexon is additionally added.  13. The method according to p. 10, characterized in that it additionally contribute to perfume, an anticorrosive substance, an antifoam agent, antifreeze, or a combination thereof.  14. The method according to claims 10 and 13, characterized in that amyl or octyl alcohol is added as the anti-foaming agent.  15. The method according to p. 13, characterized in that sodium nitrite or disodium phosphate is introduced into the mixture as an anti-corrosion agent.  16. The method according to p. 13, characterized in that glycerol or ethylene glycol is added as antifreeze.

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