RU2635476C1 - Bicillin-1 as means for prevention and early therapy of acute injuries caused by sulphur mustard - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: application of the official medicinal product of penicillin - bicillin-1 as means for prevention and early therapy of acute lesions caused by the resorptive action of sulfur mustard is proposed.

EFFECT: compared to the prototype, bicillin-1 has equal protective efficiency with respect to the resorptive action of sulfur mustard, but it is used in 20 times lesser dose and 5 times smaller volume with a more optimal way of administration, which indicates its advantage in case of mass use in preventive and early treatment.

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Description

The invention relates to the field of medicine and pharmacology, and relates to a means of prevention and early treatment of acute lesions caused by the resorptive action of sulfur mustard, which can be used in emergency situations (local conflicts, industrial accidents at chemical weapons destruction facilities).

Sulfur mustard refers to poisons of the polytropic type of action that can interfere with metabolic processes at various levels.

A large number of domestic and foreign works are devoted to the study of the mechanisms of action of sulfur mustard and the development of drugs for the prevention and early treatment of lesions caused by them [1, 2]. However, their comprehensive analysis made it possible to establish that there are still not only antidote drugs, but also pathogenetic therapy agents that are quite effective in case of damage by this vesicant and suitable for rapid widespread use.

The relevance of the search for drugs for the prevention and treatment of mustard injuries is determined by the preservation of the probability of using sulfur mustard gas as a combat and sabotage poisonous substance, as well as the possibility of toxicant damage to personnel storing and destroying chemical weapons in the production process. We should not forget about the massive flooding (over 300 thousand tons) of chemical warfare agents in various parts of the Baltic Sea that have been there since the end of World War II. According to experts, the probability of a voluminous (volley) release of toxic chemicals (including mustard gas) into the environment increases every year, because metal barrels and the design of shells and bombs are heavily corroded.

An analysis of domestic and foreign developments on finding funds for effective prevention and early treatment of acute lesions with sulfur mustard allowed us to conclude that sodium thiosulfate is recommended as a medicine for systemic use for these purposes. In this case, there is no unequivocal opinion on this drug. So, some researchers recommend the use of sodium thiosulfate only prophylactically. The possibility of its effective use for medicinal purposes is described [3]. The effectiveness of sodium thiosulfate is closest to the claimed tool and adopted as a means of the prototype.

Significant disadvantages of the prototype tool include the fact that in order to achieve the desired effect, it must be administered only intravenously and in doses approaching the maximum tolerated [2, 4]. This circumstance significantly limits the possibility of widespread use of the prototype in clinical practice, and even more so in emergency situations.

The purpose of the invention is the expansion of the list of pharmacological prophylaxis and early treatment of acute injuries with sulfur mustard due to the use of a medicinal product that has similar efficacy as compared to the prototype, which is an official drug, which is widely used in outpatient and clinical practice and is available for mass application .

This goal is achieved by using as a means of prevention and early treatment of acute lesions caused by the resorptive action of sulfur mustard, the drug penicillin-bicillin-1, which is a domestic official drug and is intended for antibacterial prevention and treatment of infectious diseases, as well as other diseases, in pathogenesis of which an important role is played by penicillin-sensitive bacteria [3, 5-7].

The theoretical prerequisites for the use of bicillin-1 for the prevention and early treatment of acute lesions with sulfur mustard are associated with the presence of sulfur atoms in the penicillin molecule that form SH groups, which help to reduce the damaging effect of sulfur mustard.

Bicillin-1 in the pharmacy network is presented in powder form in vials for the preparation of a solution for intramuscular and subcutaneous administration.

In domestic and foreign publications there is no information on the use of bicillin-1 as a means for the prevention and early treatment of acute lesions with sulfur mustard.

The ability to achieve the objectives of the invention is confirmed by the data of the experiments presented in the examples.

Example 1. A comparative study of the effectiveness of the prototype and the claimed funds according to the criterion of survival of experimental animals in acute lesions resulting from the action of sulfur mustard.

The experiments were performed on outbred white male rats weighing 180-220 g, obtained from the Rappolovo nursery RAMS (St. Petersburg). Before the experiment, all experimental animals were kept under quarantine under standard vivarium conditions for 14 days. During the experiment, three groups were formed from experimental animals using the randomization method: the first experimental (10 individuals), the second experimental (6 individuals) and control (6 individuals).

Systemic administration of sulfur mustard was carried out intramuscularly in a volume of 0.5 ml / 100 g body weight. To obtain a working solution of sulfur mustard, immediately before the experiment, a solution of dimethyl sulfoxide (DMSO) in distilled water was added to the mother liquor so that the final concentration of DMSO was 4%.

The prototype agent (sodium thiosulfate) and the inventive agent (bicillin-1) were dissolved in an ex tempore isotonic sodium chloride solution.

Sodium thiosulfate was used in the form of a 30% solution in ampoules (Novosib-Khimpharm OJSC, Novosibirsk), which was diluted with isotonic sodium chloride in a ratio of 1: 2 and was administered to experimental animals of the first experimental group at a dose of 2500 mg / kg, intraperitoneally in a volume of 2 5 ml / 200 g body weight.

The inventive tool was administered to experimental animals of the second experimental group subcutaneously in the back in a volume of 0.5 ml / 100 g of body weight. In this case, the dose of the claimed drug was 200 thousand units / kg (which corresponds to 125 mg / kg [4]).

No experimental drugs were administered to experimental animals of the control group.

In the course of the experiment, the effectiveness of early treatment 1 minute after the acute lesion arising from intramuscular injection of sulfur mustard in a dose of 4.25 mg / kg was evaluated by the change in the survival of experimental animals by the 14th day of treatment.

The criterion for the effectiveness of the prototype and the claimed funds was the increase in the survival of experimental animals.

During the statistical processing of the controlled indicators in the experimental groups, their main descriptive statistics were calculated: average value, standard deviation, median, interquartile range and mode. The normal distribution hypothesis was checked using the Kolmogorov-Smirnov criterion, and the equality of the general variances was checked using the Leven criterion. Since the distribution of values in the variational series corresponded to the normal one and the variance of the distributions were equal, when comparing the indicators, the statistical significance of the differences with the corresponding control group was evaluated using the Student t-test. The differences were considered statistically significant, in which the indicator of the level of statistical significance was p≤0.05. For each of the continuous values, data in the format M (mean value) ± m (standard error of the mean value) were used.

The research results presented in table 1 indicate that the survival rate of experimental animals by the 14th day of observation after the introduction of both the prototype and the inventive means is almost the same and is, respectively, 90.0 ± 10.0% and 100.0- 17.0%, which is 1.9 times and 2.0 times higher than the control group, the animals of which the drugs were not administered. Moreover, in comparison with the prototype agent, the inventive agent was administered in a 20-fold lower dose, subcutaneously and in a 5-fold lower volume, which significantly reduces the burden on the body.

Example 2. A comparative study of the effectiveness of the prototype and the claimed funds in their prophylactic use for the treatment of lesions with sulfur mustard.

The experiments were performed on outbred white male rats weighing 180-220 g, obtained from the Rappolovo nursery of the Russian Academy of Medical Sciences. (St. Petersburg). Before the experiment, all experimental animals were kept under quarantine under standard vivarium conditions for 14 days. During the experiment, three groups were formed from the experimental animals using the randomization method: the first experimental (10 individuals), the second experimental and control (6 animals each).

Systemic introduction of sulfur mustard was carried out as described in example 1.

Before the experiment, the prototype agent and the inventive agent were dissolved in an isotonic sodium chloride solution ex tempore.

As in example 1, sodium thiosulfate was used in the form of a 30% solution in ampoules, diluted with an isotonic sodium chloride solution in a 1: 2 ratio, and the experimental animals were introduced in a dose of 200 mg / kg, intraperitoneally, in a volume of 2.5 ml / 100 g body weight.

Experimental animals of the second experimental group were injected with the claimed agent subcutaneously in the back in a volume of 0.5 ml / 100 g body weight, at a dose of 200 thousand units / kg (which corresponds to 125 mg / kg).

No experimental drugs were administered to experimental animals of the control group.

The effectiveness of the prevention of damage to experimental animals with sulfur mustard gas with the introduction of the prototype or the claimed drug 1 hour before the intramuscular injection of sulfur mustard in a dose of 4.25 mg / kg was evaluated by the survival criterion of the experimental animals up to 14 days after exposure to sulfur mustard.

The criterion for the effectiveness of the studied drugs was the increase in the survival of experimental animals. Statistical processing of controlled parameters in the experimental groups was carried out as described in example 1. Statistically significant were considered the differences for which the indicator of the level of statistical significance was p≤0.05. For each of the continuous values, data in the format M (mean value) ± m (standard error of the mean value) were used.

It was experimentally established (table 2) that prophylactic administration 1 hour before acute exposure to sulfur mustard gas of both the prototype and the claimed drug leads to 100% and 83% survival of animals, respectively, of the first and second experimental groups at 50% death in the control group. Moreover, in comparison with the prototype agent, the inventive agent was administered in a 20-fold lower dose, subcutaneously and in a 5-fold smaller volume, which indicates its advantage.

Example 3. A comparative assessment of the effectiveness of the prototype and the claimed funds in the early treatment of acute lesions with sulfur mustard in terms of protection coefficient.

The experiments were performed on outbred white male rats weighing 180-220 g, obtained from the Rappolovo nursery RAMS (St. Petersburg). Before the experiment, all experimental animals were kept under quarantine under standard vivarium conditions for 14 days. During the experiment, four groups of 8 animals each were formed from experimental animals using the randomization method: three experimental and control.

Systemic introduction of sulfur mustard was carried out as described in example 1.

The prototype tool and the claimed tool were dissolved in an isotonic sodium chloride solution ex tempore.

The prototype agent (sodium thiosulfate) was used in the form of a 30% solution in ampoules, diluted with an isotonic sodium chloride solution in a ratio of 1: 2 and was administered to experimental animals of the first experimental group at a dose of 2500 mg / kg, intraperitoneally, in a volume of 2.5 ml / 100 g of body weight.

The inventive tool was administered to experimental animals of the second experimental group subcutaneously in the back in a volume of 0.5 ml / 100 g body weight, at a dose of 200 thousand units / kg (which corresponds to 125 mg / kg).

The animals of the third experimental group were injected with isotonic sodium chloride solution.

The animals of the control group were not injected with drugs.

The criterion for the effectiveness of the studied drugs was the protection coefficient (K ₃ ), calculated as the ratio of the average lethal dose (LD ₅₀ ) in the corresponding experimental group to LD ₅₀ in the control group. Calculation of LD ₅₀ was carried out by the method of probit analysis according to V. B. Prozorovsky [8].

The data presented in table 3 indicate that the isotonic sodium chloride solution used as a solvent for the studied drugs did not affect their toxicity when administered for therapeutic purposes. The coefficient of protection (K _S ) was the same as in the control group of animals to which drugs were not administered.

Using the prototype immediately after systemic administration of sulfur mustard allows maximum pronounced detoxifying effect (thus _{I O} was 1.36). It should be noted that the observed effect is possible with the introduction of the prototype in sufficiently large doses, comprising about half of the LD ₅₀ .

In the case of using the inventive tool provides the same high level of therapeutic effect as in the case of using the means of the prototype. The value of K _{W is} also 1.36. Moreover, it is essential that the claimed tool in comparison with the prototype tool was administered in a 20 times lower dose, subcutaneously and in a 5-fold smaller volume.

Thus, it was experimentally established that the claimed tool (bicillin-1) compared with the prototype tool (sodium thiosulfate) has equal protective effectiveness against the resorptive action of sulfur mustard, but is used in a much lower dose and volume with a more optimal route of administration, which testifies to its advantage if necessary, mass use for preventive purposes and with early treatment.

The claimed invention meets the criterion of "novelty", since for the first time for effective prevention and early treatment of acute lesions with sulfur mustard, it was proposed to use an official drug - bicillin-1, which is usually used in laboratory and clinical practice for antibacterial prevention and treatment of infectious diseases, as well as other diseases in the pathogenesis of which penicillin-sensitive bacteria play an important role.

The claimed invention meets the criterion of "inventive step", since there is no information in the well-known and accessible sources of information indicating the feasibility of using bicillin-1 as a means of prevention and early treatment of acute lesions with sulfur mustard and the possibility of achieving high efficiency of the specified official drug with subcutaneous administration in a significantly lower dose and volume.

The compliance of the claimed invention with the criterion of "suitability for use" is confirmed by the results of the above examples, which showed that the prophylactic and therapeutic efficacy of bicillin-1 is not inferior to the sodium thiosulfate widely used for these purposes, but it can be administered subcutaneously and in a significantly lower dose and volume, which is important if necessary, mass application in emergency situations. For the prevention and treatment of acute lesions with sulfur mustard, it is proposed to use an affordable pharmacy drug that is widely used in medicine.

BIBLIOGRAPHY

1. Nitrogen mustard exposure of murine skin induces DNA damage, oxidative stress and activation of MAPK / Akt-AP1 pathway leading to induction of inflammatory and proteolytic mediators / D. Kumar [et al.] // Toxicol. Lett. 2015. Vol. 235. P. 161-171.

2. Topical nitrogen mustard exposure causes systemic toxic effects in mice / D.G. Goswami [et al.] // Experimental and Toxicologic Pathology. 2015. Vol. 67, No. 2. P. 161-170.

3. Drug prophylaxis of early manifestations of acute resorptive action of alkylating substances / Methodical recommendations / Federal State Budgetary Institution of Science "Institute of Toxicology of the Federal Medical and Biological Agency", M., 2012, 44 pp.

4. Mashkovsky M.D. Medicines Manual for doctors. M .: "New wave", p. 770-774.

5. Belikov V.G. Pharmaceutical Chemistry. Graduate School. 1985.S. 717.

6. Kalmykov A.A., Aminev P.M., Magomedov M.M., Polyakov B.C. The experience of bicillin prophylaxis of acute respiratory diseases in the training center // VMZh, 2012, No. 2, p. 16-18.

7. Internal diseases. Field therapy. Ed. A.L. Rakova and A.E. Sosyukina. St. Petersburg: Tome, 2003, p. 153.

8. Prozorovsky VB A practical guide on the accelerated determination of average effective doses and concentrations of biologically active substances. Baykalsk, 1994, 46 p.

Claims (1)

The use of penicillin - bicillin-1 - as a means of prevention and early treatment of acute lesions caused by the resorptive action of sulfur mustard.