RU2431477C1 - METHOD OF GASTRIC ERADICATION OF Helicobacter pylori INFECTION - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to gastroenterology, and can be used for gastric eradication of Helicobacter pylori infection. That is ensured by using a surface-active material benzyldimethyl[3-(miristoylamino)propyl]ammonium chloride - C₂₆H₄₇CIN₂O.

EFFECT: invention provides gastric eradication of Helicobacter pylori ensured by the absence of resistance of Helicobacter pylori bacterial culture strains to benzyldimethyl[3-(miristoylamino)propyl]ammonium chloride.

2 cl, 2 dwg, 2 tbl

Description

The invention relates to medicine, namely to gastroenterology, and can be used to eradicate Helicobacter pylori infection in the stomach.

A known method of eradicating Helicobacter pylori in the stomach, based on the use of tablet forms of drugs, mainly antibiotics, in various combinations, for example, using two antibiotics in the form of tablets in a standard dosage (clarithromycin + amoxicillin or clarithromycin + metronidazole) and antioxidant secretion drug. Instead of the aforementioned triple regimen, the so-called “quadrotherapy” is used, which in addition to agents that inhibit HCl secretion include the same (clarithromycin, metronidazole) or previously unused (tetracycline) antibiotics and bismuth-containing drugs (Ivashkin V.T., Isakov V.A. Russian Railways of Gastroenterology, Hepatology, Coloproctology. 2001, No. 3, pp. 1-11).

Most of the eradication schemes used in medical practice should ensure the eradication of Helicobacter pylori infection in at least 80% of cases with tight control over the regimen of medication. However, in outpatient conditions, as a rule, the percentage of eradication does not exceed 60-65%.

Currently, in Russia more than 15-20% of Helicobacter pylori strains are resistant to clarithromycin, and resistance to metronidazole has reached almost 60%. Amoxicillin is the only antibiotic to which H. pylori does not develop resistance (Zakharova N.V. Consilium medicum. Gastroenterology, Appendix Issue No. 2, 2004, pp. 7-10).

The closest analogue to that claimed by the authors is the method of eradication Helicobacter pylori, which consists in the fact that drugs in powder form are mixed with orange juice in a ratio of 1.0 g per 90-100 ml and use this solution 2 times a day for half an hour before administration food (RF Patent No. 2253449, A61K 31/43, 2005) for 7 or 4 days. The powdered form of drugs contributes to their faster dissolution in orange juice. Orange juice containing citric acid slows the evacuation of fluid from the stomach, thereby increasing the interaction time of drugs with the surface of the mucous membrane. In addition, the organic components of the juice, having a pronounced sorption ability, firmly bind to the luminal surface of the mucus layer, neutralize its charge and, eliminating hydrophobic properties, facilitate the diffusion of the drug solution deep into the layer, that is, into the area of the infect.

Clinical testing of the method of eradication (liquid technology) proposed in the RF Patent №2253449 was carried out with a 7-day course of treatment for 100 patients with chronic gastritis and peptic ulcer, as well as for 50 similar patients with a 4-day course. The aforementioned four-component treatment regimen was used: famotidine, de-nol, nifuratel or furazolidone, amoxicillin (Flemoxin solutab). In both modes (4 or 7 days), 100% eradication of Helicobacter pylori infection was achieved, while when using a similar scheme in tablets, the eradication does not exceed 80-90%.

The aim of the invention is the creation of a new highly effective and economical method for the treatment of gastroenterological diseases initiated by the bacteria Helicobacter pylori.

The technical result in the proposed method is achieved in that for the eradication of Helicobacter pylori bacteria in the stomach instead of three or four drugs, only one bactericidal surfactant is used - benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride - C ₂₆ H ₄₇ ClN ₂ O , the production of which has been mastered by domestic pharmaceutical enterprises. The bactericidal agent C ₂₆ H ₄₇ ClN ₂ O is used as a solution in water and / or in food mineral or organic diluents at a concentration of ₂₆ H ₄₇ ClN ₂ O in solutions from 0.0025 to 0.01%.

Benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride - and its 0.01% aqueous solution are approved for medical use and are registered by the Ministry of Health of the Russian Federation under registration numbers 91/146/1 and 91/146/2. Production was also mastered and registered in the Rospotrebnadzor of the Russian Federation (No. 77.99.1.2. U 7641.9.08) a 15% solution of C ₂₆ H ₄₇ ClN ₂ O-Antisept-15 (TU 9392-03-95548551-08).

The effect of the drug on the bacteria Helicobacter pylori was studied on three strains of these bacteria: laboratory 26695 (for which the nucleotide sequence of the genome is known) and two clinical isolates A37 and A45 isolated from patients with chronic gastritis. Clinical strains at the beginning of the studies withstood no more than 15 passages on a solid nutrient medium. Helicobacter pylori was cultured on a solid nutrient medium: Columbian agar (Biomerieux, France), 10% FBS (HyClone, USA), 5% yeast extract, at 37 ° C under microaerophilic conditions with passivation after 48-72 hours. Into a liquid nutrient medium Helicobacter pylori culture was transferred by suspending cells grown on plates in physiological saline and plating the resulting bacterial suspension in BHI medium (Biometrieux, France) containing 10% FBS and 5% yeast extract. We used a Helicobacter pylori culture that withstood at least two passages in a liquid medium. The daily culture of Helicobacter pylori was transferred after centrifugation into physiological saline (0.9% NaCl) containing various amounts of benzyl dimethyl [3- (myristoylamino) propyl] ammonium chloride, and incubated for 5 ° C in an atmosphere of 5% CO ₃ for 48 hours. The number of surviving bacteria was estimated by seeding on a solid nutrient medium serial dilutions of the suspension. As a control, a suspension of Helicobacter pylori in physiological saline not containing benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride was used.

Counting the number of grown colonies was carried out on the fourth day. The experiments were performed in triplicate. The results are presented in Table 1.

Table 1 Growth of Helicobacter pylori after incubation in a solution of benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride The concentration of benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride in a solvent Strain 26695 Strain A37 Strain A45 0.0001% + + + 0,0002% - - - 0,0005% - - - 0.001% - - - 0.002% - - - 0.005% - - - 0.01% - - - Note: “-” - lack of bacterial growth "+" - bacteria growth is noted

To obtain a solution of benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride, distilled water, physiological saline (0.9% NaCl), table mineral water, and natural fruit and vegetable juices were used.

It was found that the results of bacterial inactivation were not affected by the diluent contained benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride, and the concentration of inactivating agent had an effect on bacterial inactivation.

Differences in inactivation between the studied strains of the bacterial culture of Helicobacter pylori were not observed, which confirms the lack of their resistance to the proposed method of bactericidal exposure and therefore the high efficiency of the new method of eradicating the infection.

An additional study of the toxicity and safety of the use of a 15% solution of С ₂₆ Н ₄₇ СlN ₂ O was carried out at the Federal State Institution Research Institute of Disinfectology of Rospotrebnadzor (R.P. Rodionova, Zh.P. Alekseeva, M.M. Maltseva. “Scientific report on the results of medical preventive disinfectant "Antisept-15", submitted for state registration ". M., 2007).

The studies were carried out on several types of laboratory animals (rats, mice, guinea pigs and rabbits), statistical groups - at least 6-10 individuals of small and large rodents. The experimental and control animals were of the same line, species, age, sex, and weight characteristics (weight of mice - 18-22 g, rats - 180-220 g, guinea pigs - 220-250 g, rabbits - 3000-3500 g).

When applying the drug C ₂₆ H ₄₇ ClN ₂ O at a dose of 2500 mg / kg to the skin of mice, neither clinical manifestations of intoxication nor death of animals were observed during the 2-week observation period.

In the stomach of rats, females and males, the drug was administered in the form of a 15% aqueous solution in the dose range: 500-1000-2000-3000 mg / kg. LD ₅₀ "Antisept 15" for female rats was 1450 mg / kg, for male rats - 1300 mg / kg. According to the specified parameter, the compound belongs to class 3 of moderately dangerous compounds, when applied to the skin of mice - to class 4 there are few dangerous compounds according to Classification GOST 12.1.007-76.

When assessing the degree of danger of the studied substance in saturating concentrations of vapors in experimental animals (mice), no symptoms of intoxication were observed, as well as their death neither during the exposure period for 60 minutes, nor on the following days of observation for 2 weeks. When examining experimental animals after exposure to vapors, no changes were found in the indicators of general toxicity relative to the control group. Thus, in terms of volatility, the compound belongs to the 4th class of low hazard according to the classification of the degree of danger of disinfectants.

A study of the local irritating effect on the skin was carried out on two types of animal rabbits and guinea pigs with a single exposure (without flushing) in concentrations of aqueous solutions of 1% -5% -15%. The investigations carried out showed that the application of Antisept 15 solutions to the skin of guinea pigs at the indicated concentrations did not cause signs of skin irritation. Rabbits were more sensitive to substances, but the reaction was the same from both substances: applications of 1% aqueous solutions did not cause signs of irritation, 5% and 15% solutions caused mild erythema (1 point according to S.V.Suvorov's classification). The introduction of a 0.1% aqueous solution of the compound into the eyes of rabbits caused only short-term blepharospasm for 15-20 seconds. In this case, there was no irritation of the mucous membranes, iris and cornea.

The main results of experiments to assess the toxicity and safety of the use of C ₂₆ H ₄₇ ClN ₂ O obtained in studies on laboratory animals are shown in Table 2.

Table 2. Indicators 15% solution of C ₂₆ H ₄₇ ClN ₂ O (Antisept 15) LD ₅₀ when introduced into the stomach of a rat, mg / kg,
Hazard Class 1450; 1300
3 LD ₅₀ when applied to the skin, mg / kg,
Hazard Class More than 2500
four Volatility, C ₂₀
Hazard Class Lack of clinic and death
Hazard class 4 Local irritant effect on the skin (once), guinea pigs None (1% -5% -15% solutions) Local irritant effect on the eyes (once) None (0.1% solution)

The clinical use of the proposed method for the eradication of Helicobacter pylori infection in the stomach should be preceded by a mandatory medical examination of the patient.

Diagnosis of Helicobacter pylori infection is usually done by interviewing a patient for dyspeptic complaints and symptoms, and then performing tests to confirm or deny the fact of Helicobacter pylori infection.

Testing can be performed using a histological or cytological examination of gastric biopsies, as well as a non-invasive HELIK ^® test system with a digital apparatus manufactured by the domestic industry for the initial diagnosis of Helicobacter pylori infection, to monitor the progress of anti-Helicobacter therapy and to verify the effectiveness of the therapy already performed.

The device is applicable in the practice of gastroenterologists, therapists, pediatricians and family doctors. The sensitivity of the HELIK ^® test system with a digital apparatus is 95%, and the specificity is 97%. The principle of the HELIK ^® test system is based on the biochemical method for determining infection with Helicobacter pylori by its urease activity, i.e. the ability to quickly hydrolyze urea.

The patient receives a urea solution of normal isotopic composition, and the gas formed during hydrolysis enters the air of the oral cavity. Using a highly sensitive sensor, the digital apparatus determines whether this gas is contained in the air of the oral cavity and presents this information on the screen. Analyzing this information, the doctor concludes that the patient is infected.

The duration of the examination is 9 minutes. An example of a positive test for the primary diagnosis of Helicobacter pylori infection is shown in FIG.

The proposed method for the eradication of bacteria Helicobacter pylori in the stomach is clinically implemented as follows:

with confirmed indications of eradication of Helicobacter pylori on an empty stomach, in the morning and evening 30-40 minutes before a meal, the patient takes 5-20 ml of a 0.1% solution of C ₂₆ H ₄₇ ClN ₂ O, additionally diluted in 100-200 ml of freshly boiled water or orange juice at room temperature;

after taking the drug, the patient needs to lie on his left side and, taking a lying position, turn every 2-4 minutes from side to side;

after 30 minutes, the patient can eat;

This eradication therapy is carried out for 3-4 days in the morning and evening. The control determination of Helicobacter pylori is carried out by the same methods at least four weeks after the course of eradication.

An example of a negative test for the re-diagnosis of Helicobacter pylori infection is shown in FIG.

Clinical observation during the year of patients showed the absence of exacerbation of diseases. The obtained results confirm an increase in the eradication efficiency when using the technology of taking solutions of the bactericidal agent C ₂₆ H ₄₇ ClN ₂ O, the possibility of a significant reduction in the duration of therapy and lower financial costs for treatment.

Claims (2)

1. A method for eradicating a Helicobacter pylori infection in the stomach, including exposure to a bactericidal agent, which is used as a surfactant benzyldimethyl [3- (myristoylamino) propyl] ammonium chloride - C ₂₆ H ₄₇ ClN ₂ O. 2. The method according to claim 1, characterized in that the bactericidal agent C ₂₆ H ₄₇ ClN ₂ O is used as a solution in water and / or in food mineral or organic diluents at a concentration of C ₂₆ H ₄₇ ClN ₂ O in solutions from 0, 0025 to 0.01%.

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