Allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({ 2-[(2,5-dimethylphenyl)amino]-2-oxoethyl} thiophosyl)-2-methyl-1,4-dihydropyridine-3-carboxylate, exhibiting analgesic properties

Abstract

FIELD: medical; pharmaceutical chemistry; pharmacology.SUBSTANCE: use of allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate of formula (I) as an analgesic agent.EFFECT: compound of formula (I) has a pronounced analgesic effect, exceeding the effect of the comparison drug - metamizole sodium.1 cl, 2 tbl, 2 ex(I)

Description

[0001] The invention relates to the field of medical and pharmaceutical chemistry, pharmacology, namely to the use of an organic compound, allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate of formula (I), as an analgesic agent with antinociceptive activity, which can be used in medicine and veterinary medicine.

[0002]

[0003] (I)

[0004] Pain syndrome, as a sign of an inflammatory reaction, is one of the leading reasons for people seeking medical help both in Russia and worldwide [Barinov A.N. Complex pain treatment / / Rus.med. jour. 2007. Vol. 15, No. 4. Pp. 215-220; Shostak N.A., Klimenko A.A., Demidova N.A. et al. Safety of selective non-steroidal anti-inflammatory drugs: analysis of recent data / / Clinician. 2020. Vol. 14, No. 1-2. Pp. 91-99]. The multifaceted impact of pain syndrome on the human body determines its independent clinical significance.

[0005] Acute and chronic pain syndromes of various origins and locations are widespread worldwide. According to clinicians, chronic pain syndrome accounts for over 88% of cases, with the majority of patients having high pain descriptors (5 points or higher on the Visual Analogue Scale). Statistics show that back pain is the most common pain symptom, accounting for 46.5% of cases. Headaches rank second, accounting for 22.6% of cases. Joint pain ranks third (21.4%). More than half a billion people worldwide suffer from osteoarthritis-related pain [Tkachenko P.E., Ivashkin V.T., Maevskaya M.V. Practical recommendations for the correction of hepatotoxicity induced by antitumor therapy / / Practical recommendations RUSSCO. Malignant tumors. 2023. T.13, Part 2. P. 69-82].Long-term chronic pain associated with osteoarthritis is a key factor in increasing the incidence of cardiovascular events.

[0006] Chronic muscle and bone pain, including back pain, joint pain, and Tietze's syndrome, is the most common type of chronic pain. A characteristic feature of such conditions is the predominance of pain in the clinical picture even after complete functional restoration of damaged tissue [Kotova O.V., Akarachkova E.S. Adverse effects of nonsteroidal anti-inflammatory drugs: possible solutions / / Consilium Medicum. 2017. No. 2. pp. 2-12].

[0007] Dorsalgia is the second most common cause of temporary disability. In Russia, almost every second patient visits a clinic due to pain, and half of these visits involve back pain. Statistics show that approximately 40% of patients with chronic nonspecific lower back pain fail to find effective pain relief.

[0008] Furthermore, chronic pain syndrome can very often be caused by headaches (migraines) and facial pain [Shostak N.A., Klimenko A.A., Demidova N.A. et al. Safety of selective non-steroidal anti-inflammatory drugs: analysis of recent data / / Clinician. 2020. Vol. 14, No. 1-2. Pp. 91-99]. In the global population, up to 15% of people suffer from migraine, while it is considered the sixth most common disease and causes significant economic damage, especially in the group of women of reproductive age.

[0009] According to statistics, 75% of patients experiencing periodic attacks of chronic pain do not seek medical help from specialists [Tkachenko P.E., Ivashkin V.T., Maevskaya M.V. Practical recommendations for the correction of hepatotoxicity induced by antitumor therapy / / Practical recommendations RUSSCO. Malignant tumors. 2023. Vol.13, Part 2. Pp. 69-82]. In this regard, the colossal difference in the rates of prescription and actual use of non-steroidal anti-inflammatory drugs (NSAIDs) becomes clear. Thus, annually 500 million patients are prescribed various NSAIDs, and about 26 billion packages of them are sold [Barinov A.N. Complex pain treatment / / Rus. med. journal. 2007. Vol.15, No.4. P. 215-220; Tkachenko P.E., Ivashkin V.T., Maevskaya M.V. Practical recommendations for the correction of hepatotoxicity induced by antitumor therapy / / Practical recommendations RUSSCO. Malignant tumors. 2023. Vol. 13, Part 2. P. 69-82].Long-term chronic pain associated with oncological diseases becomes a prerequisite for the development of pronounced anxiety, depressive disorders, aggression, insomnia and even suicidal intentions [Kotova O.V., Akarachkova E.S. Adverse effects of non-steroidal anti-inflammatory drugs: possible options for overcoming the situation / / Consilium Medicum. 2017. No. 2. pp. 2-12].

[0010] A current trend in modern medical and pharmaceutical science is the development of issues related to the introduction of domestic highly effective painkillers and anti-inflammatory drugs with a high safety profile.

[0011] To meet the needs for effective and safe pain relief and anti-inflammatory treatment, active development and study of new drugs with a favorable benefit / risk ratio is required.

[0012] Alpha-cyanothioacetamide (CAS 7357-70-2) is a promising reagent for the creation of structural analogs exhibiting a wide spectrum of biological activity. Partially hydrogenated derivatives of α-cyanothioacetamide, which have high analgesic and anti-inflammatory activity, are of great interest. These properties determine the promise of new compounds in terms of their biological activity [Ketova E.S., Bibik E.Yu., Batishcheva G.A., Krivokolisko S.G. Comprehensive assessment of the hepatoprotective activity of new derivatives of alpha-cyanothioacetamide / / Far Eastern Medical Journal. 2024. No. 1. pp. 22-27; Bibik E.Yu., Oleinik I.S., Pankov A.A., Frolov K.A., Dotsenko V.V., Krivokolisko S.G. Study of new 1,4-dihydropyridine derivatives as potential agents with analgesic activity in the orofacial trigeminal pain test: an experimental preclinical randomized study / / Kuban Scientific Medical Bulletin. 2023. No. 30 (2). P.64-75; Bibik I.V., Pankov A.A., Frolov K.A., Dotsenko V.V., Krivokolisko S.G., Bibik E.Yu. Study of hematotoxicity of new derivatives of condensed 3-aminothieno[2,3-b]pyridines and 1,4-dihydropyridines with high analgesic activity / / Avicenna Bulletin. 2023. No. 4. P. 499-507].

[0013] The problem to be solved by the invention is to establish analgesic activity among new derivatives of α-cyanothioacetamide.

[0014] The problem is solved by using allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate (I) (IUPAC name – allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate) as an analgesic agent, which has more pronounced antinociceptive properties in comparison with the reference drug – metamizole sodium.

[0015] Compound (I) was obtained at the Khimex Research Laboratory of the Volodymyr Dahl Luhansk State University using the procedure described for similar compounds [Krivokolysko DS, Dotsenko VV, Bibik E.Yu., Samokish AA, Venidiktova Yu.S., Frolov KA, Krivokolysko SG, Vasilin VK, Pankov AA, Aksenov NA, Aksenova IV New 4-(2-Furyl)-1,4-dihydronicotinonitriles and 1,4,5,6-Tetrahydronicotinonitriles: Synthesis, Structure, and Analgesic Activity / / Russian Journal of General Chemistry. 2021. Vol. 91. No. 9. P. 1646-1660; Krivokolysko DS, Dotsenko VV, Bibik E.Yu., Samokish AA, Venidiktova Yu.S., Frolov KA, Krivokolysko SG, Pankov AA, Aksenov NA, Aksenova IV New Hybrid Molecules Based on Sulfur-Containing Nicotinonitriles: Synthesis, Analgesic Activity in Acetic Acid-Induced Writhing Test, and Molecular Docking Studies / / Russian Journal of Bioorganic Chemistry. 2022. Vol. 48. No. 3. P. 628-635; Tilchenko DA, Bibik E.Yu., Dotsenko VV, Krivokolysko SG, Frolov K.A., Aksenov N.A., Aksenova I.V. Synthesis and Hypoglycemic Activity of New Nicotinonitrile-Furan Molecular Hybrids / / Russian Journal of Bioorganic Chemistry. 2024. Vol. 50. No. 2. P. 554-570].

[0016] The most similar in structure to the described compound are various biologically active substances that served as objects for the creation of analgesic, hypotensive, antiallergic, and diuretic drugs [Gouda MA, Hussein BH, Helal MH et al. A Review: Synthesis and Medicinal Importance of Nicotinonitriles and Their Analogous / / Journal of Heterocyclic Chemistry. 2018. Vol. 55(7). P. 1524-1553; Rucins M., Plotniece A., Bernotiene E. et al. Recent Approaches to Chiral 1,4-Dihydropyridines and their Fused Analogues / / Catalysts. 2020. Vol. 10(9). P. 1019; Ling Y., Hao ZY, Liang D. et al. The Expanding Role of Pyridine and Dihydropyridine Scaffolds in Drug Design / / Drug Design, Development and Therapy. 2021. Vol. 15. P. 4289-4338; Salem MA, Helel MH, Gouda MA et al. Overview on the synthetic routes to nicotine nitriles / / Synthetic Communications Reviews. 2018. Vol. 48(4). P. 345-374].

[0017] For the first time, it was established that the biologically active substance allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate (I) exhibits analgesic activity in an experiment on laboratory animals (white outbred sexually mature male rats) when compared with a drug from the group of non-steroidal anti-inflammatory drugs - metamizole sodium, surpassing the reference sample.

[0018] Pharmacological properties of the claimed compound.

[0019] Example 1.

[0020] Our preliminary virtual bioscreening and predictive analysis according to [Yang J., Kwon S., Bae Park SH et al. Galaxy Sagittarius: Structure- and Similarity-Based Prediction of Protein Targets for Druglike Compounds / / Journal of Chemical Information and Modeling. 2020. Vol. 60. P. 3246-3254; Gfeller D., Grosdidier A., ​​Wirth M. et al. SwissTargetPrediction: a web server for target prediction of bioactive small molecules / / Nucleic Acids Research. 2014. Vol. 42(1). P. 32-39; Gfeller D., Michielin O., Zoete V. Shaping the interaction landscape of bioactive molecules / / Bioinformatics. 2013. Vol. 29. P.3073-3079] showed that the mechanism of action of allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate (I) is associated with the fact that its molecules with a high degree of probability act on such receptors as arachidonate-5-lipoxygenase, cyclooxygenase-2 and adenosine receptors A1, A2a, A2b, and, as a result, are potentially capable of reducing pain sensitivity while providing anti-inflammatory, antipyretic and antiexudative effects [Bocheva A.A., Krivokolisko S.G. Study of antipyretic activity of new partially hydrogenated pyridines, derivatives of α-cyanothioacetamide / / Actual problems of experimental and clinical medicine. Collection of articles of the 81st international scientific and practical conference of young scientists and students. Volgograd. 2023. Pp. 425-426; Bibik E.Yu., Bushik A.A., Samokish A.A., Venediktova Yu.S., Krivokolisko D.S. et al.Efficacy of new 1,4-dihydrothiopyridine derivatives in the orofacial trigeminal pain test / / Safety of pharmacotherapy: Noli Nocere! Collection of abstracts of the IV All-Russian scientific and practical conference with international participation. Kazan. 2021. pp. 19-21].

[0021] Example 2.

[0022] The experimental study was carried out on sexually mature outbred male rats weighing 280±10 g obtained from the vivarium of the Federal State Budgetary Educational Institution of Higher Education “Lugansk State Medical University named after St. Luke” of the Ministry of Health of the Russian Federation (FGBOU VO “LSMU named after St. Luke of the Ministry of Health of the Russian Federation) in the conditions of a specialized laboratory of the Department of Fundamental and Clinical Pharmacology of the Federal State Budgetary Educational Institution of Higher Education “LSMU named after St. Luki Ministry of Health of the Russian Federation in accordance with the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines and rules for working with animals based on the provisions of the Helsinki Declaration and recommendations contained in EU Directive 86 / 609 / ECC and the Council of Europe Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes, as well as in accordance with the rules of good laboratory practice when conducting preclinical studies in the Russian Federation (Article 11 of Federal Law No. 61-FZ of 12.04.2010 "On the circulation of medicines", with the order of the Ministry of Health of the Russian Federation dated April 1, 2016 No. 199n (On approval of the Rules of Good Laboratory Practice) and GOST 33044-2014 "Principles of Good Laboratory Practice", approved by Order of the Federal Agency for Technical Regulation and Metrology No. 1700-st dated November 20, 2014).

[0023] The research was approved by the Bioethics Committee of the Federal State Budgetary Educational Institution of Higher Education “Lugansk State Medical University named after St. Luke” of the Ministry of Health of the Russian Federation (Russian Federation, Lugansk, apt. 50-letiya Oborony Luganska, 1g), protocol No. 5 dated 12 / 10 / 2022.

[0024] Ratification into groups was performed randomly using the "envelope method." Animals were divided into a control group (with simulated orofacial trigeminal pain and a hot plate test), a reference group (rats receiving the comparison drug, metamizole sodium), and an experimental group (animals receiving the original α-cyanothioacetamide derivative, compound of formula I). ​​Each group consisted of eight animals of the same sex.

[0025] The animals were kept in identical conditions and received food and water freely.

[0026] The analgesic activity of the new cyanothioacetamide derivative was determined using two tests, in accordance with the recommendations for preclinical studies [Guidelines for experimental preclinical studies of new pharmacological substances / Ed. G. U. Khabriev. - M .: Medicine. - 2005. - 832 p.]:

[0027] a) orofacial trigeminal pain modeled by subcutaneous injection of 0.1 ml of 5% formalin solution into the vibrissae of experimental animals. The analgesic effect was assessed by recording the number of scratching movements of the forepaws in the orofacial area per minute, which was counted at 10-, 15-, and 20-minute intervals after the injection of 0.1 ml of 5% formalin solution into the vibrissae;

[0028] b) Hot plate. Animals were placed on a metal plate heated to 50-55ºC (average 52ºC), surrounded by a glass cylinder. The time from the moment the rats were placed on the hot plate to the onset of jumping, licking, and hind paw withdrawal, reflecting the behavioral response to the corresponding nociceptive stimulation, was recorded.

[0029] The condensed cyanothioacetamide derivative under study was administered intragastrically at a dose of 5 mg / kg of animal body weight 1.5 hours before acute pain was simulated in vivo. Metamizole sodium, administered intragastrically at a dose of 7 mg / kg of animal body weight, served as the reference drug.

[0030] The indicators of analgesic effect were: a decrease in the number of scratching movements in the orofacial trigeminal pain test and a reliable increase in the latent period of the reaction after the introduction of the test substance in the hot plate test.

[0031] The analgesic activity of allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate was compared with that of the reference drug, metamizole sodium (analgin).

[0032] For statistical evaluation of the obtained data during the study, we used Microsoft Excel 2019, an online calculator for calculating statistical criteria https: / / medstatistic.ru / calculators.html, and the licensed computer program Statistica 10.0 for Windows.

[0033] In the process of assessing biological parameters in experiments on outbred laboratory animals, the use of non-parametric methods of statistical data processing is recommended due to the abnormal nature of the sample distribution [Lugovik I.A., Makarova M.N. Toxicological studies. Reference intervals of mass coefficients of internal organs in a sample consisting of 1000 outbred rats. Laboratory animals for scientific research. 2021. P. 1. https: / / doi.org / 10.29296 / 2618723X-2021-01-01].

[0034] The homogeneity and reliability of the experimental data were assessed using the coefficient of variation (V) and the Wilcoxon signed-rank (W) test. If the coefficient of variation is less than 10%, the variability of the data series is considered insignificant.

[0035] When processing the experimental data, the following were determined: the arithmetic mean of the indicators (quantitative indicators) – t; the dispersion of the values ​​σ 2 around the arithmetic mean, standard deviation – σ and coefficient of variation – V, %.

[0036] The data from experimental studies of analgesic activity in two pharmacological tests are presented in Tables 1 and 2.

[0037] Table 1. Results of the pharmacological test of orofacial trigeminal pain to identify the analgesic activity of a new original derivative of cyanothioacetamide

[0038] Groups Statistical characteristics: quantitative indicators (t), dispersion σ2, standard deviation σ, variation coefficient V, % 10 minutes after the introduction of algogen (5% aqueous formaldehyde solution) 15 minutes after the introduction of algogen (5% aqueous formaldehyde solution) 20 minutes after the introduction of algogen (5% aqueous formaldehyde solution) Control t = 100.6; σ2 = 271; σ = 16.4; V = 16.4 t = 70.7; σ2 = 68.2; σ = 8.2; V = 11.7 t = 57.2; σ2 = 265; σ = 16.3; V = 28.4 Reference (metamizole sodium) t = 31; σ2 = 41.6; σ = 6.45; V = 20.8 t = 22.8; σ2 = 32.7; σ = 5.7; V = 25.1 t = 9.2; σ2 = 13.8; σ = 3.71; V = 40.3 Experimental (new derivative of cyanothioacetamide (I)) t = 31.2; σ2 = 130; σ = 11.4; V = 36.4 t = 14.8; σ2 = 22.1; σ = 4.7; V = 31.7 t = 4.5; σ2 =2.8; σ = 1.6; V = 36.8

[0039] Evaluating the data presented in Table 1, it can be concluded that in the orofacial trigeminal pain modeling test, the experimental compound reduces the frequency of scratching movements by 3.2 times compared to the control group 10 minutes after the administration of algogen; by 4.8 times compared to the control group 15 minutes after the administration of algogen; by 12.7 times compared to the control group 20 minutes after the administration of algogen.

[0040] The studied compound from among the new cyanothioacetamide derivatives was 2 times better than the results obtained in the group receiving the drug metamizole sodium.

[0041] Table 2. Duration of stay on the hot plate of experimental rats of the experimental groups

[0042] Groups Statistical characteristics: quantitative indicators (t) of the duration of stay on the hot plate, dispersion σ2, standard deviation σ, variation coefficient V, % Control t = 1.80 σ2 = 0.21; σ = 0.46; V = 25.2 Reference (metamizole sodium) t = 2.84 σ2 = 0.12; σ = 0.34; V = 12.1 Experimental (new derivative of cyanothioacetamide (I)) t = 6.87 σ2 = 46.3; σ = 6.81; V = 99.1

[0043] As can be seen from the data presented in Table 2, the duration of stay on the hot surface upon introduction of a new compound from among the cyanothioacetamide derivatives was 6.87 seconds, which is 3.82 times longer than in rats of the control group, and 2.5 times longer than upon introduction of the comparison drug – metamizole sodium.

[0044] Thus, the pharmacological correction of acute pain syndrome, modeled in two classical pharmacological tests on white rats, by a new derivative of cyanothioacetamide – allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate (I), is characterized by a high degree of antinociceptive activity, exceeding that of the comparison drug – metamizole sodium.

[0045] No adverse events were recorded during experimental studies.

[0046] Summarizing the above statistical data and conclusions, the described facts and observations clearly demonstrate the ability of allyl 4-[4-(benzyloxy)-3-methoxyphenyl]-5-cyano-6-({2-[(2,5-dimethylphenyl)amino]-2-oxoethyl}thio)-2-methyl-1,4-dihydropyridine-3-carboxylate (I) to exhibit pronounced analgesic activity in an in vivo experiment in orofacial trigeminal pain and hot plate tests. Moreover, in terms of the degree of expression of analgesic properties, the test compound exceeded the comparison drug (metamizole sodium) by 2.0-2.5 times.

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