RU2736936C1 - 4,10-di(ethoxyacetyl)-2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo[5,5,0,03,11,05,9] dodecane as an analgesic agent and a method for production thereof - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to compound - 4,10-di(2-ethoxyacetyl)-2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo[5,5,0,0^3,11,0^5,9]dodecane. Invention also relates to a method for its preparation, which includes acylation of 2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo[5,5,0,0^3,11,0^5,9]dodecane with 2-ethoxyacetic acid chloride.

EFFECT: obtaining a novel compound of formula (I), having analgesic activity, which can be used in medicine.

2 cl, 4 tbl, 2 ex

Description

The invention relates to pharmaceutical chemistry and pharmacology, namely to a new chemical substance - 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5, 5.0.0 ^3.11 , 0 ^5.9 ] dodecane, which has analgesic activity:

In accordance with the wording of the International Association for the Study of Pain (IASP), pain is an unpleasant sensory sensation and emotional experience associated with true or potential tissue damage or described in terms of such damage [Bondarenko D.A., Dyachenko I.A., Skobtsov D.I., Murashev A.N. In vivo models for studying analgesic activity. Biomedicine. 2011, 2: 84-94; Mashkovsky M.D. Medicines: 15th ed. M .: OO "New Wave Publishing House". 2008, 1206; Guidelines for conducting preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., ZAO Grif and K. 2012]. Pain as an integrative function of the body is a negative biological need responsible for the formation of a functional system for maintaining homeostasis. Depending on the leading pathogenetic factor of development, all pain syndromes were divided into three main groups: nociceptive, neurogenic and psychogenic. Many acute and chronic illnesses, injuries and medical interventions involve pain that requires the use of analgesics. According to the IASP definition (1994), pain is classified as acute and chronic. The choice of anesthetics of the appropriate analgesic potential is determined by a number of factors, which include etiology, intensity, type of pain, and individual characteristics of the patient. Currently, for the treatment of pain of various origins in inpatient and outpatient settings, the following groups of analgesics are mainly used: non-opioid analgesics and opiates, their various combinations in multimodal therapy regimens, analgesics from the cannabinoid class [Mashkovsky M.D. Medicines: 15th ed. M .: OO "New Wave Publishing House". 2008, 1206; Mikhailova A.S. The analgesic arsenal of the clinician. Pharmateca. 2018, (3): 50-56; Sosnov A.V., Sadovnikov S.V., Semchenko F.M., Rufanov K.A., Tokhmakhchi V.N., Sosnova A.A., Tyurin I.A. Potent non-narcotic analgesics as a direction in the development of pharmaceuticals. Development and registration of medicines. 2016, 14 (1): 196-206; Cawich S.O., Deonarine U., Harding H.E., Dan D., Naraynsingh V., Cannabis and Postoperative Analgesia. Handbook of Cannabis and Related Pathologies. Biology, Pharmacology, Diagnosis, and Treatment. 2017, 450-458]. Non-opioid analgesics, which are non-narcotic drugs and are available without a prescription, are used to treat acute pain and chronic pain syndromes of low intensity [Mikhailova A.S. The analgesic arsenal of the clinician. Pharmateca. 2018, (3): 50-56]. Non-opioid analgesics include drugs of five pharmacological groups: NSAIDs, selective inhibitors of cyclooxygenase-2 (COX-2), pyrazolone derivatives, derivatives of paraacetaminophenol and flupirtine. Indications for the appointment of opioid analgesics are acute and chronic pain of moderate to high intensity, not eliminated by non-opioid analgesics.

There are contraindications and restrictions to the appointment of most analgesics [Mashkovsky M.D. Medicines: 15th ed. M .: OO "New Wave Publishing House". 2008, 1206; Mikhailova A.S. The analgesic arsenal of the clinician. Pharmateca. 2018, (3): 50-56; Guidelines for conducting preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., JSC "Grif and K". 2012 .; Sosnov A.V., Sadovnikov SV., Semchenko F.M., Rufanov K.A., Tokhmakhchi V.N., Sosnova A.A., Tyurin I.A. Potent non-narcotic analgesics as a direction in the development of pharmaceuticals. Development and registration of medicines. 2016, 14 (1): 196-206; Cawich S.O., Deonarine U., Harding H.E., Dan D., Naraynsingh V., Cannabis and Postoperative Analgesia. Handbook of Cannabis and Related Pathologies. Biology, Pharmacology, Diagnosis, and Treatment 2017, 450-458].

One of the most frequently used and the closest in technical result to the proposed tool is Tramadol - an analgesic with a mixed mechanism of action, including opioid and non-opioid components, which is selected as a comparison drug [Mashkovsky M.D. Medicines: 15th ed. M .: OO "New Wave Publishing House". 2008, 1206; Mikhailova A.S. The analgesic arsenal of the clinician. Pharmateca. 2018, (3): 50-56].

We have revealed for the first time a pronounced analgesic activity of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane belonging to the 3rd hazard class "low-toxic agents".

The problem solved by the present invention is to expand the arsenal of analgesics with the help of a newly synthesized substance.

The task is achieved by using the first synthesized substance 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane as an analgesic agent. The technical result of the proposed invention is a substance, a method for its preparation and the identification of the analgesic effect of a new non-toxic analgesic agent.

The inventive analgesic agent 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane was synthesized for the first time at the Institute for Problems of Chemical and Energy Technologies (Biysk).

In the Federal State Budgetary Institution “Research Institute of Pharmacology named after E.D. Goldberg "SB RAMS (Tomsk) during experimental studies revealed a pronounced analgesic activity of this substance.

The new property of the means proposed as an invention does not explicitly follow from the prior art in this field and is not obvious to a specialist.

The proposed invention can be used in medicine. An identical set of features was not found in the study of the state of the art in the patent and scientific medical literature [Tolstikova T.G., Morozova E.A., Sysolyatin S.V., Kalashnikov A.I., Zhukova Yu.I., Surmachev V. N. Synthesis and biological activity of derivatives 2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3,11, 0 ^5,9] dodecane. Chemistry for Sustainable Development. 2010, 18: 511-516; Sergey V. Sysolyatin, Alexander I. Kalashnikov, Valeriy V. Malykhin, Irina A. Surmacheva, Gennady V. Sakovich. Reductive Debenzelation of 2,4,6,8,10,12 - Hexaazaisowurtzitane / International Journal of Energetic Materials and Chemical Propulsion. 2010, 9 (4): 365].

Based on the foregoing, the claimed technical solution should be considered as corresponding to the criteria: "Novelty", "Inventive level", "Industrial applicability".

The possibility of implementing the claimed invention is confirmed by an example of a specific implementation of the method for obtaining the claimed agent.

Example 1

Preparation of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaaza-tetracyclo [5,5,0,0 ^3.11, 0 ^{5, 9} ] dodecane

14.71 g (0.12 mol) of 2-ethoxyacetic acid chloride was dissolved in 100 ml of dry acetonitrile and transferred to a flask with a stirrer and reflux condenser. Then was added 10.00 g (0.03 mol) of 2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5, 9} ] dodecane. The suspension was boiled with stirring for 5 hours, filtered and cooled to room temperature. Acetonitrile was distilled off under vacuum, and the residue was washed with 3 × 30 ml of methanol. Received 13.47 g of product with a basic substance (HPLC) of 96.7%. The resulting product (13.47 g) was dissolved by heating in a mixture of 135 ml of ethyl alcohol and 36 ml of aceonitrile. The resulting solution was cooled to room temperature. The precipitated product was filtered off, washed with 3 × 30 ml of water. Air dried.

Received 11.94 g (78%) 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^{3 , 11} , 0 ^5.9 ] dodecane in the form of a colorless crystalline product with a basic substance content of 99.99% (by HPLC) and a melting point of 230.5-231.5 ° C.

C ₂₂ H ₃₂ N ₆ O ₈ Found (%) C 51.89; H 6.31; N 16.49. Calculated (%): C 51.96; H 6.34; N 16.53; About 25.17.

IR spectrum, v / cm ^-1 : 3045, 2985, 2931, 2889, 1678, 1665, 1651, 1412, 1360, 1285, 1169, 1135, 979.715, 702, 620.

¹ H NMR spectrum (DMSO-d6, δ, ppm): 1.09-1.17 (t, 6H, CH ₃ ), 1.97-2.07 (m, 12H, CH ₃ CO), 3.35 (s, 4H, CH ₂ ), 3.38-3.52 (q, 4H, CH ₂ ), 4.25-4.65 (m, 2H, CH), 6.30-6.94 (m, 4H, CH).

¹³ C NMR spectrum (DMSO-d6, δ, ppm): 15.38 (C, CH ₃ , EtO), 21.29, 22.42 (C, CH ₃ , CH ₃ CO); 40.40, 62.33, 63.75 (C, CH), 66.04 (C, CH ₂ , EtO), 68.10, 69.24 (C, CH ₂ ), 69.60, 71.89, 73.91, 167.72, 168.26, 169.29 (C, CO, CH ₃ CO ).

The possibility of implementing the claimed invention is confirmed by an example of a specific study of the analgesic activity of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0, 03,11,05,9] dodecane in comparison with the reference drug Tramadol, which proves that the new signs solve the problem.

Example 2

The experiments were performed on 62 outbred CD-I mice of both sexes, 50 male CBA mice (7-8 weeks old, weight 22-27 g) of the first category of conventional ones obtained from the Department of Experimental Biomodelling of the N.I. E. D. Goldberg of the Tomsk NIMTs (animal health certificate, veterinary certificate 270 No. 0008633). The keeping of animals and the design of experiments were approved by the Bioethical Committee of the N.I. E. D. Goldberg and were compliant with Directive 2010/63 / EU of the European Parliament and of the Council of the European Union on the protection of animals used for scientific purposes [Directive 2010/63 / EU of the European Parliament and of the Council of the European Union on the protection of animals used for scientific purposes (translated from English. ). SP. 2012, 48]; Order of the Ministry of Health of the Russian Federation of August 1, 2016 N 199n.

Each animal within the group was assigned an individual number from 1 to 10, labeled with fucorcin. Mice were randomly assigned to groups, using body weight as a criterion, so that the individual weight did not deviate from the mean within one sex by more than ± 10%. The data (weight and number) of the selected animals were ranked in descending order of weight using an Excel computer program with the SORT command.

Statistical processing of the obtained results was carried out by the methods of variation statistics using the Statistica 6.0 software. For all data, the mean value (X) and standard error of the mean (m) were calculated, which, together with the value n (the number of variants in the group), are presented in the summary tables. The difference between the compared values was considered significant if the probability of their identity was less than 5% (P <0.05). Using the sample coefficients of asymmetry and kurtosis, the degree of approximation of the distribution law of the studied trait to the normal one was estimated. In cases of normal distribution of signs, the parametric Student's t-test was used for statistical evaluation. In case of deviations of the distributions of the trait from the normal form for independent samples, the nonparametric U-Wilcoxon-Mann-Whitney test was used. To identify the reliability of differences in qualitative indicators used the criterion of angular transformation of Fisher [Guidelines for preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., ZAO Grif and K. 2012].

Determination of acute toxicity of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5, 9} ] dodecane (I).

When determining acute toxicity, substance (I) was injected into 6 outbred female mice and 6 outbred male CD-I mice once intragastrically at a maximum dose of 2000 mg / kg (due to poor water solubility 6.25 g / L) at 0, 5 ml of purified water. During 2 weeks of observation, the death of the animals was not observed, the mice had no abnormalities in health and behavior. Due to the impossibility of determining LD _{50, the} test substance (I) is assigned to the 3rd hazard class - “low-toxic agents” in accordance with GOST 12.1007-76 “Harmful substances. Classification and general safety requirements "and" Guidelines for the conduct of preclinical studies of drugs, 2012 "[Guidelines for the conduct of preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., CJSC "Grif and K". 2012].

For screening assessment of the effect of the test substance on the development of pain reaction (I) under the conditions of the "Acetic writhing" model and the "Mechanical compression of the paw" test [Guidelines for conducting preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., JSC "Grif and K". 2012; Barrot M. Tests and models of nociception and pain in rodents. Neuroscience. 2012,211: 39-50] was administered to mice via a gavage in the dose range of 25, 50, 100 and 200 mg / kg once daily for 3 days. The used daily dose of the reference drug Tramadol for mice (10 mg / kg) corresponded to the average daily dose for humans (according to the instructions for using Tramadol). Animals of the negative control group received the solvent (purified water) in a similar route of administration. The body weights of the animals were used to calculate the doses of the test substances. The selection of a certain amount of the test substance was carried out in a working room with a bactericidal recirculator, which ensures the safety of procedures and the cleanliness of the preparation of the test substances for administration. The required amount of substances for injections was prepared by a laboratory employee responsible for pharmacological work. Dose preparation was based on dose values and indicated administration volumes.

The study of the analgesic activity of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5, 9} ] dodecane (I) in comparison with the reference drug Tramadol in the "Acetic cramps" test (Abdominal constriction test).

Test "Acetic cramps" (Abdominal constriction test) is aimed at the study of acute visceral and somatically deep pain [Guidelines for preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., JSC "Grif and K". 2012; Barrot M. Tests and models of nociception and pain in rodents. Neuroscience. 2012, 211: 39-50]. A specific pain reaction - "cramps" (characteristic movements of animals, including contractions of the abdominal muscles, alternating with their relaxation, stretching of the hind limbs and arching of the back) - were caused by intraperitoneal administration of 0.75% solution of acetic acid (GOST 61-75) to mice at a dose of 0 , 1 ml / 10 g of body weight. The analgesic effect was assessed by the ability of a potential drug (within 20 min after injection) to reduce (in%) the number of "writhing" as compared to the control group of animals (the criterion of effectiveness was a decrease in the pain response by at least 50%) and the latent time of the onset of pain reactions. At the end of the experiments, the animals were euthanized by cranio-cervical dislocation.

In this series of experiments, the following groups of outbred male CD-I mice were used:

1. Negative control - mice were injected with purified water in a volume of 0.5 ml / mouse daily intragastrically through a tube for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% solution of acetic acid (n = 10) ;

2. Positive control - mice were injected with Tramadol at a dose of 10 mg / kg in a volume of a solvent of 0.5 ml / mouse daily intragastrically through a tube for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% acetic solution acids (n = 8);

3. Analgesic - mice were injected (I) at a dose of 25 mg / kg in a volume of solvent 0.5 ml / mouse daily intragastrically through a tube for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% solution acetic acid (n = 8);

4. Analgesic - mice were injected (I) at a dose of 50 mg / kg in a volume of 0.5 ml / mouse daily intragastrically through a tube for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% solution acetic acid (n = 8);

5. Analgesic - mice were injected (I) at a dose of 100 mg / kg in a volume of 0.5 ml of solvent / mouse daily intragastrically through a tube for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% solution acetic acid (n = 8);

6. Analgesic - mice were injected (I) at a dose of 200 mg / kg in a volume of 0.5 ml / mouse intragastrically through a tube daily for three days, on the last day the administration was carried out 1 hour before the injection of 0.75% solution acetic acid (n = 8).

The number of vinegar "writhings" 27.11 ± 4.85 and the latent time of their onset 337 ± 75 s indicate the development of a full-fledged pain reaction in animals of the negative control group after the introduction of a chemical stimulus (Table 1). A three-day course of administration of Tramadol to mice at a therapeutic dose of 10 mg / kg led to a decrease in the level of pain response by 39.1%, since the number of "writhing" in this group was statistically significantly reduced by 1.6 times compared with the corresponding indicator in the negative control group. The latent time of the development of pain reaction did not have statistically significant differences relative to the indicator of the negative control group. The obtained results of the reference control correspond to the literature data.

The use of substance (I) at a dose of 25 mg / kg contributed to a decrease in the severity of the pain reaction by 44.7% as a result of a decrease in the number of vinegar "writhing" by 1.8 times (P <0.05) relative to the same indicator in animals treated with purified water (Table 1).

In the case of the introduction of (I) at a dose of 50 mg / kg, its maximally pronounced analgesic effect was noted in comparison with all experimental groups, since the value of inhibition of the pain reaction was 86.6% (Table 1). In addition, the longest time of development of the pain reaction was recorded, 622 ± 143, relative to the corresponding indicators of all experimental groups (table 1).

A further increase in the dose of a potential analgesic led to a gradual decrease in the effect. If, as a result of the introduction of (I) at a dose of 100 mg / kg, the analgesic effect was 48.4% (P <0.05), then an increase in the dose to 200 mg / kg contributed to a decrease in the effectiveness to 37.3 (P> 0.05) ...

Thus, the test substance 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^5.9 ] dodecane (I) in the "Acetic cramps" test (Abdominal constriction test) exhibits pronounced analgesic activity at doses of 25, 50 and 100 mg / kg. In the groups of animals that received the new substance, the absence of writhing was found in 25% (50 mg / kg) and 12.5% (200 mg / kg) of animals, which was not observed in the group of Tramadol. Substance (I) at a dose of 50 mg / kg has a predominant activity (P <0.05) compared to the reference drug Tramadol (suppression of the pain reaction was 86.6% versus 39.1%, respectively).

The study of the analgesic activity of 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5, 9} ] dodecane (I) in comparison with the reference drug Tramadol in the Mechanical Paw Compression test (Randall-Selitto test).

When investigating the analgesic activity of a new molecule (I) in the Mechanical Paw Compression test, an analgesimeter (Ugo Basile, Italy) is used, which is designed to perform accurate testing of analgesics on normal and inflamed paws of rats and mice using the Randall-Selitto method [ conducting preclinical studies of drugs. Ed. Mironova A.N., Bunatyan N.D. et al. M., JSC "Grif and K". 2012; Barrot M. Tests and models of nociception and pain in rodents. Neuroscience. 2012, 211: 39-50]. With an increase in pressure on the limb, the following were consistently observed: the spinal reflex of withdrawing the limb, the complex of supraspinal motor reactions of the animal aimed at releasing the limb, and vocalization. The threshold of vocalization (weight in grams), latent time of pain response, and the number of animals without pain response were recorded. This type of mechanical stimulation has certain disadvantages: 1) sometimes it is difficult to accurately measure the intensity of the stimulus; 2) repeated mechanical irritation can lead to a decrease or increase in sensitivity; 3) low sensitivity of the method requires the use of high intensity effects.

The criterion of the analgesic effect was considered a significant decrease in the intensity of pain reactions, estimated in g by the force of the effect on the healthy paw, the latent time of the pain reaction, the number of animals without a pain reaction in dynamics (1, 2 hours after the administration of the drugs) after a single and course three-day administration. Euthanasia of animals at the end of the experiments was carried out by cranio-cervical dislocation.

In this series of experiments, the following groups of male CBA mice were used:

1. Negative control - mice were injected with purified water in a volume of 0.5 ml / mouse daily intragastrically through a tube for three days; on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latent time of pain reaction, number of animals without pain reactions (n = 10);

2. Positive control - mice were injected with Tramadol at a dose of 10 mg / kg in a volume of a solvent of 0.5 ml / mouse daily intragastrically through a tube for three days, on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latent time of pain reactions, number of animals without pain reaction (n = 8);

3. Analgesic - mice were injected (I) at a dose of 25 mg / kg in a volume of a solvent of 0.5 ml / mouse daily intragastrically through a tube for three days, on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latent time pain response, number of animals without pain response (n = 8);

4. Analgesic - mice were injected (I) at a dose of 50 mg / kg in a volume of 0.5 ml of solvent / mouse daily intragastrically through a tube for three days, on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latent time pain response, number of animals without pain response (n = 8);

5. Analgesic - mice were injected (I) at a dose of 100 mg / kg in a volume of 0.5 ml solvent / mouse daily intragastrically through a tube for three days, on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latency time pain response, number of animals without pain response (n = 8);

6. Analgesic - mice were injected (I) at a dose of 200 mg / kg in a volume of 0.5 ml solvent / mouse daily intragastrically through a tube for three days, on the first and last day of the experiment, the administration was carried out 1 hour before fixation of vocalization, latency time pain reaction, the number of animals without pain reaction (n = 8).

On the Randall-Selitto model of mechanical hyperalgesia, the pain threshold (r) was in the first (367.6 ± 77.4; 406.1 ± 59.1) and (370.4 ± 68.6; 326.3 ± 62, 2) the third day of observation in animals of the negative control group (table 2, 3).

The reference drug Tramadol statistically significantly increased the threshold of pain sensitivity 1 hour after administration with a single use, 2 hours (P <0.05) after administration during the course application (table 2, 3).

In animals that received substance (I) at doses of 25, 50 and 200 mg / kg with a single application, there was a tendency to an increase in the threshold of pain sensitivity 1 hour after administration to 1.6; 1.4 and 1.6 times relative to control, respectively. The maximum analgesic activity was observed in the group using a dose of 100 mg / kg (P <0.05). There was no statistically significant difference between the above data and that of the Tramadol group. A similar direction of effects was observed 2 h after the administration of the test substance (I) during this observation period, and the potential analgesic showed pronounced analgesic activity at doses of 25 mg / kg (P <0.05) and 100 mg / kg (P <0.05 ) (table 2, 3). The absence of statistically significant results after 1 hour (25, 50 and 200 mg / kg) and 2 hours (50 and 200 mg / kg) after the introduction of substance (I) is due to the large scatter of data within the experimental groups, which may be due to the individual sensitivity of the animals. ...

A statistically significant increase in the pain sensitivity threshold in response to the action of an increasing mechanical stimulus, pointwise applied to the hind leg of the mouse, was found in the group of animals that received substance (I) at a dose of 100 mg / kg 1 hour after administration as a result of its course use (Table 3 ). In the remaining groups of substance use, there was a tendency to an increase in the pain threshold. The reference drug Tramadol showed similar activity (P> 0.05).

In the last observation period (2 hours after administration with course use), a statistically significant increase in the threshold of pain sensitivity was recorded in relation to the following groups: Tramadol and substance (I) at doses of 100 and 200 mg / kg (Table 3). It should be noted that the effects of the reference drug and the test substance are comparable.

Based on the totality of the results obtained for the pain threshold indicators, it can be concluded that the predominant activity of substance (I), which demonstrated statistically significant activity during all periods of observation, mainly at a dose of 100 mg / kg, in contrast to Tramadol.

An important indicator of the analgesic activity of new drugs is the latent time of the development of pain reaction (table 2, 3). 1 and 2 hours after a single administration of Tramadol, there was a tendency to an increase in latency time (scatter of data within the group). In animals that received substance (I) at doses of 25, 50 and 200 mg / kg, there was also a tendency to an increase in this indicator in the first hour of observation with a single application, which may well be due to the individual sensitivity of the animals. The pronounced analgesic activity of the tested analgesic (I) at a dose of 100 mg / kg, judging by a statistically significant increase in the latency time (1.8 times) relative to the negative control indicator, was detected 1 hour after its administration during this observation period. 2 hours after administration, the maximum value of the latent time of development of the pain response was in the group using substance (I) at doses of 25 and 100 and mg / kg relative to the corresponding values of the negative control group (Table 2). It should be noted that there is no statistically significant difference with Tramadol.

Course use of both Tramadol and substance (I) increased their effectiveness after 1 and 2 hours of observation. Thus, in the group of administration of an opioid analgesic, a statistically significant increase in the time of development of pain reaction was revealed (Table 3) in the dynamics of this stage of observation. In the case of the use of substance (I) at doses of 25 mg / kg, 100 mg / kg and 200 mg / kg, a significant increase in the index of 1 hour (P <0.05) and 2 hours (P <0.05) was recorded; 1 hour (P <0.05) and 2 hours (P <0.01); 2 h (P <0.05), respectively (table 3).

Thus, the analysis of the indicators of the latent time of the development of the pain reaction allows us to conclude about the predominant activity of substance (I), which demonstrated statistically significant activity during all periods of observation, mainly at a dose of 100 mg / kg, in contrast to Tramadol.

One of the criteria characterizing the analgesic activity of the test substance may be the number of animals with the maximum manifestation of the analgesic effect - exposure for 15 s on the device (Table 4).

At the first stage of observation (with a single use) among animals of all studied groups, a statistically significant number of mice with the maximum analgesic effect (15 s with the exposure period on the device) was observed when using (I) at a dose of 100 mg / kg (2 h; 62.5% ; P <0.05). The course use of the tested substances led to a statistically significant increase in the number of animals without signs of pain in all study groups in the first hour of observation (Table 4). After 2 h of observation, the maximum value of 50% of mice without signs of pain was noted in the group using substance (I) at a dose of 100 mg / kg, while in other groups there was a trend. It should be noted that the studied indicator of pain syndrome (I) at a dose of 100 mg / kg was statistically significant not only relative to the corresponding value of the negative control, but also Tramadol.

Thus, as a result of the study of the analgesic activity of substance (I) in the test "Mechanical compression of the paw" (Randall-Selitto test) in male CBA mice with single and three-day use in dynamics (1, 2 hours after administration), a pronounced antinociceptive activity in the dose range of 25-200 mg / kg. It should be noted that the maximum effect, predominant in comparison with Tramadol, was revealed after the introduction of substance (I) at a dose of 100 mg / kg.

Analysis of the results obtained in the study of the analgesic activity of 4,10-d and (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane (I) in comparison with the reference drug Tramadol in the tests "Acetic cramps" (Abdominal constriction test) and "Mechanical compression of the paw" (Randall-Selitto test) allows us to conclude that the newly obtained substance (I) has a pronounced analgesic effect in the dose range of 25-200 mg / kg, exceeding in a number of indicators the activity of the reference drug Tramadol.

The data obtained on the pronounced analgesic effect of the low-toxic 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane (I) allow us to conclude that the study of the selected compound as an analgesic agent for the relief of moderate and high intensity pains of various origins and, to a certain extent, predict the nature of its analgesic effect in humans.

Claims (2)

1. 4,10-di (2-ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5 , 9} ] dodecane (I), which has analgesic activity. 2. A process for preparing 4,10-di (ethoxyacetyl) -2,6,8,12-tetraacetyl-2,4,6,8,10,12-geksaazatetratsiklo [5,5,0,0 ^3.11, 0 ^{5 , 9} ] dodecane (I) according to claim 1, which consists in the acylation of 2,6,8,12-tetraacetyl-2,4,6,8,10,12-hexaazatetracyclo [5,5,0,0 ^3,11 , 0 ^5.9 ] dodecane with 2-ethoxyacetic acid chloride.