RU2780247C1 - Application of a tricyclic sulphur-containing derivative of 1,2-dihydroquinoline for inhibiting the replication of betacoronaviruses, including sars-cov-2 - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to the field of pharmacology and medicine, namely, to an application of a tricyclic sulphur-containing derivative of 1,2-dihydroquinoline by the general formula (I) for inhibiting the replication of a betacoronavirus, including SARS-CoV-2. Compounds by the formula (I) exhibit low toxicity and can be used to treat diseases caused by betacoronaviruses, including SARS-CoV-2. In the general formula (I), R1 constitutes a substituent selected from a methyl, ethyl, methoxy, ethoxy group; R2 constitutes a substituent selected from a methyl, ethyl, methoxy, ethoxy group; R3 constitutes a substituent selected from a methyl, ethyl, methoxy, ethoxy group; R4 constitutes a substituent selected from a methyl, benzyl, 3-phenylacryloyl, 3-(4-methoxyphenyl)acryloyl, 3-phenylpropanoyl, phenylethyl, 2-(4-fluorophenoxy)acetyl, 4-chlorophenoxyacetyl, 2-(2,4-dichlorophenoxy)acetyl, 4-nitrobenzoyl, 2-(1,3-dioxoisoindoline-2-yl)-3-phenylpropanoyl) group; 3,4-dichlorophenylcarbamoyl, furan-2-carbonyl, ethylcarbamoyl, 2-naphthylyloxyacetyl, 2-(1,3-dioxoisoindoline-2-yl)acetyl, 2-(1,3-dioxohexahydro-1H-isoindole-2(3H)-yl)acetyl, 2-morpholinoacetyl, piperidine-1-ylacetyl, 2-(4-carbamoylpiperidine-1-yl)acetyl, 2-(2,5-dioxopyrrolidine-1-yl)acetyl, 2-phenoxyacetyl, 2-1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)acetyl, 2-benzo[d]oxazole-2-ylthio)acetyl, 2-(benzo[d]thiazole-2-yloxy)acetyl, benzoxazole-2-ylmercaptoacetyl, furanoyl, 2-naphlinylyoxymethylcarbonyl, 3,4-dichlorophenylcarbamoyl; X constitutes a substituent selected from sulphur, unsubstituted or substituted in positions 2,3,4,5,6-arylamine selected from a group including 4-(4-chlorobenzyloxy)phenylimino, 4-isopropylphenylimino, 4-(6-methylbenzo[d]thiazole-2-yl)phenylimino, 4-(N-benzoylsulphamoyl)phenylimino, 4-ethoxyphenylimino, 4-ethoxycarbonylphenylimino, phenylimino, 4-chlorphenylimino, 3-chlorphenylimino, 4-sulphamoylphenylimino, 4-(N-acetylsulphamoyl)phenylimino, 4-(N-(thiazole-2-yl)sulphamoyl)phenylimino, 2-nicotinoylhydrazono, 4-((4-chlorobenzyl)oxy)phenylimino, 2-methoxyphenylimino, 4-(N-(pyrimidine-2-yl)sulphamoyl)phenylimino, 4-(N-(4,6-dimethylpyrimidine-2-yl)sulphamoyl)phenylimino.

EFFECT: effective inhibition of the replication of betacoronaviruses, including SARS-CoV-2, characterised by high efficiency of inhibiting the replication of coronavirus in the Vero E6 cell culture at low values of the effective EC50 concentration, low cytotoxicity, and high value of selectivity index SI.

4 cl, 1 dwg, 1 tbl, 20 ex

Description

Technical field

The invention relates to the field of pharmacology and medicine, namely, to the use of a known compound - a derivative of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione of formula (I) for the treatment of diseases caused by betacoronaviruses, including SARS-CoV-2. The invention can be used for the production of a pharmaceutical composition for the treatment of these diseases.

State of the art

The worldwide outbreak of COVID-19 caused by SARS-CoV-2 and its associated consequences is a threat to the public health and economies of many countries. The lack of specific therapy against the new virus requires the search for drugs that counteract the infection.

Coronaviruses (Coronaviridae) are a large family of RNA viruses that can infect humans and some animals. The modern classification includes four groups - alpha, beta, gamma and delta coronaviruses [Helena Jane Maier, Erica Bickerton, Paul Britton / Coronaviruses: An Overview of Their Replication and Pathogenesis // Coronaviruses. - 2015; 1282; 1-23]. The genus Betacoronavirus is considered the most dangerous to humans. In humans, coronaviruses can cause a range of illnesses, from mild forms of acute respiratory infection to severe acute respiratory syndrome. In particular, three coronaviruses from the known β-coronaviruses - SARS-CoV-1, MERS-CoV and SARS-CoV-2 (the source of the COVID-19 pandemic) cause more severe symptoms and lead to a higher mortality rate. At the same time, there are no specific highly effective therapeutic agents that exhibit high biological activity in inhibiting the reproduction of beta-coronavirus.

There is an intensive search for therapeutic agents around the world to prevent and treat COVID-19. In this regard, the World Health Organization and national ministries of health are issuing constantly updated guidelines for the prevention, diagnosis and treatment of coronavirus infection using existing therapeutic agents previously intended for other viral infections (influenza, AIDS, Ebola, etc.).

The most widely used drug in the treatment of COVID-19 today is Favipiravir (FVP) 19_v7.pdf], where 6-fluoro-3-hydroxy-2-pyrazinecarboxamide was used as the active substance, as well as Remdesivir (RMD) [https://nypost.com/2020/05/01/fda-approves-remdesivir- as-emergeney-coronavirus-treatment/], and Aprotinin (APR) [A. Azimi.TMPRSS2 inhibitors, Bromhexine, Aprotinin, Camostat and Nafamostat as potential treatments for COVID-19, 10.31226/osf.io/a3rvm].

Known is the use of dalargin (hexapeptide diacetate) for the treatment of coronavirus infection COVID-19 [RU 2728939, 2020], which is part of commercially available drugs previously used for the treatment of gastric and duodenal ulcers, pancreatitis and pancreatic necrosis. The drug can be used in the form of inhalations or injections, while the drug contains dalargin in an amount of 0.1 to 50 mg per dose.

It is known the use of statins used to lower and control cholesterol levels, reduce mortality, the likelihood of development and severity of cardiovascular accidents - such as myocardial infarction and stroke, in the treatment of coronavirus infection [Patent CN 111603465, 2020; CN 111632053, 2020; CN 111588720, 2020; CN 111617065, 2020; CN 111588719, 2020]. The drug is in the form of a gel, soft capsule, oral preparation, injection, lyophilized powder for injection or solution for infusion.

However, the hitherto known compounds and agents proposed for the treatment of COVID-19 have several disadvantages that prevent their widespread use. Thus, in experiments on Vero E6 cell cultures, Favipiravir showed an almost complete inability to suppress the replication of the SARS-CoV-2 coronavirus, since its effective concentration of 50% suppression of the replication of this coronavirus is only EC50 = 61.88 μM, and the selectivity index (the ratio of cytotoxicity to EC50 ) is very low SI > 6.46 [M. Wang, R. Cao. L. Zhang, X. Yang, J. Liu, M. Xu, Z. Shi, Z. Hu, W. Zhong, G. Xiao / Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro / / cell res. - 2020; thirty; 269-271. https://doi.org/10.1038/s41422-020-0282-0]; in addition, favipiravir has shown low treatment efficacy in clinical trials, and there is no reliable evidence for the absence of side effects for it yet [Annoor Awadasseid, Yanling Wu, Yoshimasa Tanaka, Wen Zhang / Effective drugs used to combat SARS-CoV-2 infection and the current status of vaccines // Biomedicine & Pharmacotherapy. - 2021; 137; 111330; https://doi.org/10.1016/j.biopha.2021.111330]. Remdesivir to increase therapeutic efficacy must be administered as intravenous infusions in the daily dose range of 100-200 mg for at least 6 days, which leads to an increase in aspartate aminotransferase and alanine transaminase. Elevated liver enzymes can be a sign of inflammation or damage to liver cells, low albumin, low potassium, low red blood cells, low clotting platelets, and yellowing of the skin. That is, Remdesavir has a fairly high cytotoxicity, does not effectively suppress the replication of the SARS-CoV-2 coronavirus, and has a low selectivity index.

Basel. - 2005. pp. 173-199]. В 2003-2015 гг. были найдены обратимые и необратимые ингибиторы основной протеазы SARS - CoV [Pillaiyar, Т., Manickam, М., Namasivayam, V., et al. / An Overview of Severe Acute Respiratory Syndrome-Coronavirus (SARS-CoV) 3CL Protease Inhibitors: Peptidomimetics and Small Molecule Chemotherapy // Journal of Medicinal Chemistry. - 2016; 59 (14); 6595-6628], однако большинство из них были слабыми ингибиторами, и ни один из них не прошел клинических испытаний. Основная протеаза и несколько других белков SARS-CoV-2 были идентифицированы в качестве терапевтической мишени для препаратов против COVID - 19. Высококачественные 3D-структуры SARS - CoV - 2 M^pro в форме а по с различными ингибиторами депонированы в Protein Data Bank [Berman, H.M., Westbrook, J., Feng, Z., et al. / The Protein Data Bank // Nucleic Acids Research. - 2000; 28 (1); 235-242]. Выявлено, что SARS-CoV-2 M^pro имеет каталитическую диаду цистеин-гистидин: Cys145 и His41. Данные сведения были использованы при поиске соединений среди известных, в качестве основных ингибиторов протеазы SARS, которые можно было бы отнести к группе соединений-кандидатов, обладающих потенциальной противовирусной биологической активностью в отношении бета-коронавирусов для дальнейшей экспериментальной проверки.A number of publications are known that present data on SARS-CoV regarding the spatial atomistic model of its main protease [Anand, K., Ziebuhr, J., Wadhwani, P., et al. / Coronavirus main proteinase (3CLpro) structure: basis for design of anti-SARS drugs // Science (New York, NY). - 2003; 300 (5626); 1763-1767], experimental crystal structure [Yang, H., Yang, M, Ding, Y., et al. / The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor // Proceedings of the National Academy of Sciences of the United States of America. - 2003; 100 (23); 13190-13195], and the key role of M ^pro in SARS-CoV and MERS-CoV replication [Anand, K., Yang, H., Barflam, M., et al. / Coronavirus main proteinase: target for antiviral drug therapy VT - Coronaviruses with Special Emphasis on First Insights Concerning SARS,

Basel. - 2005.pp. 173-199]. In 2003-2015 reversible and irreversible inhibitors of the main SARS protease - CoV have been found [Pillaiyar, T., Manickam, M., Namasivayam, V., et al. / An Overview of Severe Acute Respiratory Syndrome-Coronavirus (SARS-CoV) 3CL Protease Inhibitors: Peptidomimetics and Small Molecule Chemotherapy // Journal of Medicinal Chemistry. - 2016; 59 (14); 6595-6628], however, most of them were weak inhibitors, and none of them passed clinical trials. The main protease and several other proteins of SARS-CoV-2 have been identified as a therapeutic target for anti- ^COVID -19 drugs. HM, Westbrook, J., Feng, Z., et al. / The Protein Data Bank // Nucleic Acids Research. - 2000; 28(1); 235-242]. SARS-CoV-2 M ^pro was found to have a cysteine-histidine catalytic dyad: Cys145 and His41. This information was used in the search for compounds among the known, as the main SARS protease inhibitors, which could be attributed to the group of candidate compounds with potential antiviral biological activity against betacoronaviruses for further experimental verification.

The disadvantage of all of the above compounds is their low efficiency of binding to the specified target, a rapid decrease in concentration with a half-life of about 1 hour and, as a result, the need to use delivered drugs at higher concentrations, which can cause adverse reactions in patients listed above.

The claimed invention is based on the use of a known compound - a derivative of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione, for the treatment of diseases caused by betacoronaviruses, incl. SARS-CoV-2, which has been shown to inhibit the replication of betacoronaviruses, and may be recommended for use in COVID-19 therapy.

Compounds are known from the prior art, which are derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione, and methods for their preparation, based on the participation of various reaction centers available in 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione molecule described below.

For the first time, the synthesis of derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones was proposed by heating 1-R-6-R'-2,2,4-trimethyl -1,2-dihydroquinolines with a 3-5-fold excess of sulfur in N,N-dimethylformamide to form the corresponding 5-R-8-R'-4,5-dihydro-4,4-dimethyl-2,3-dithiolo[3 ,4-c]quinoline-1-thiones with a yield of 15-50% [Brown, J. P. // J. Chem. soc. C. - 1968; 1074. Patent FR 1429915 (A), 1966]. The main disadvantage of this method is the rather low yield of target products. Further, it was found that an increase in the yield of reaction products up to 45-55% can be achieved by replacing the previously mentioned dihydroquinolines with similar 2,2,4-trimethyl-1,2,3,4-tetrahyroquinolines and by reacting with sulfur when heated in trichlorobenzene [Shikhaliev Kh. S., Kasaikina O.T., Shmyreva Zh. V. // Bulletin of the Academy of Sciences of the USSR Division of Chemical Science. - 1989; 38 (1.2); 176 - 178].

The preparation of derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones was proposed to be carried out in two stages through the formation of the S-methyl derivative iodide followed by interaction with n-phenethidine by boiling in ethanol for about 6 hours [Brown, J. P. // J. Chem. soc. C. - 1968; 1074-1075]. Further, this reaction was extended to a number of primary amines: aniline, ethylamine, cyclohexylamine [Patent GB 1174830 (A), 1969]. As a result of this interaction, 1,2-dithiol-3-imines were obtained. The main disadvantage of these methods is the use of a very limited number of amines for the synthesis of derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones. In [Medvedeva S.M., Zubkov F.I., Yankina K.Yu., Grudinin D.G. / Reaction of substituted 1-methylthio-4,5-dihydro[1,2]dithiolo[3,4-c]-quinoline iodides with arylamines. Synthesis of novel -pyrrolo[3,2,1-ij]quinoline-4,5-diones // Arkivoc. - 2017; III; 269-278] presents the possibilities of synthesis of (8-R-7-R'-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene) (4(2)-R''-phenyl)amines with a yield of 69-81% as a result of interaction with various arylamines by boiling in a mixture of isopropyl alcohol and pyridine for 2-3 hours.

During N-acylation of 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thiones with anhydrides or acid chlorides of acrylic, benzoic and acetic acids at boiling in anhydrous toluene in within 6 hours, the formation of 5-R-4,5-dihydro-4,4-dimethyl-2,3-dithiolo-[3,4-c]quinoline-1-thiones occurs. Carrying out N-acylation under the previously indicated conditions is also possible when using 4-chloro-3-nitrobenzoyl chloride or nicotinoyl chloride as an acylating agent [Shikhaliev, Kh.S., Shmyreva Zh.V., Zalukaev L.P. / Acylation of 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione // Zh. organ, chemistry. - 1988; 24(1); 232-233]. In this work, it is shown that similar structures can be obtained not only by modifying 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione, but at the stage formation of a heterocyclic matrix due to the reaction of substituted quinolines, for example, 1-(2,2,4-trimethylquinolin-1(2H)-yl)ethan-1-one or 1-benzoyl-2,2,4-trimethyl-1,2 -dihydroquinoline by boiling with sulfur in N,N-dimethylformamide for 11 hours. The main disadvantage of this method is the use of a limited number of acylating agents and the high temperature of the process. N-Acylation of 8-R-4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione can be carried out with chloroacetyl chloride in toluene medium with a product yield of 65% [ X.S. Shikhaliev, S.M. Medvedev, V.V. Pigarev, A.S. Solovyov, G.V. Shatalov / New heterocyclic compounds based on 8-R-4,4-dimethyl-2,3-dithiolo[5,4-c]quinoline-1-thiones // Zh. total chemistry. - 2000; 70(3); 484-486]. For the resulting 8-R-N-chloroacetyl-4,4-dimethyl-2,3-dithiolo[5,4-c]-quinoline-1-thione, the possibility of carrying out a subsequent substitution reaction in the interaction with heterocyclic mercaptans, including 1,3 -benzoxazole-2-thiol, 1,3-benzothiazole-2-thiol, 4,5-dihydrothiazol-2-thiol, 2-mercapto-4-phenylthiazole-5(4H)-thione in the presence of potassium carbonate in medium 1.4 -dioxane at 40-50°C [Shikhaliev H.S., Pigarev V.V., Solovyov A.S., Shatalov G.V. / Synthesis of N-(2-hetarylacetyl)-8-R-4,5-dihydro-4,4-dimethyl-2,3-dithiolo[5,4-c]quinoline-1-thiones // Izv. universities. Chemistry and chem. technology. - 2000; 43(2); 93-94]. The disadvantages of this method include a limited number of derivatives obtained.

The benzene ring acts as another reactive center in 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thiones, where the nitration reaction can be realized with a mixture of nitrogen ( ρ=1.4 g/cm ³ ) and acetic acids, which leads to the formation of 4,4-dimethyl-8-nitro-4,5-dihydro-[1,2]dithiolo[3,4-c]quinoline-1-thiones [Kasaikina O.T., Golovina N.A., Shikhaliev Kh.S., Smyreva Zh.V. / Kinetic description of the oxidation of hydrocarbons inhibited by sulfur-containing hydrogenated quinolines // Russian Chemical Bulletin. - 1994; 43 (5); 755 - 759. OT Kasaikin, HA Golovina, Zh.V. Shmyreva, XS Shikhaliev / Phenomenology of hydrocarbon oxidation inhibition by sulfur-containing hydrogenated quinolines // Izv. Academy of Sciences of the USSR. Ser. chem. - 1994; 5; 814-818]. The possibility of alkylating 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione with triphenylmethanol with the formation of 4,4-dimethyl-8 -trityl-4,5-dihydro-[1,2]dithiolo[3,4-c]quinoline-1-thione. The main disadvantage of this method is the use of a limited number of acylating and alkylating agents and the process at high temperatures for a long time. Nitration into the benzene ring can be realized by reaction with acetyl nitrate in chloroform to form 6-nitro-[1,2]dithiolo[3,4-c]quinoline-1-thione. [Shikhaliev Kh.S., Pigarev V.V., Shmyreva Zh.V., Medvedeva S.M., Ermolova G.I. / Nitration of 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione and its derivatives // Izvestiya VUZov. Chemistry and chemical technology. - 1998; 41; 48-51].

It was shown that 8-alkyl-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones, N-acyl-[1,2]dithiolo [3,4-c]quinoline-1-thiones, N-hetaryl(thio)acetyl-[1,2]dithiolo[3,4-c]quinoline-1-thiones have antimicrobial and antifungal activity. Among the bacteria tested, En. cloacae, followed by S. aureus, and L. monocytogenes were the most resistant. Among the fungi, T. viride was the most sensitive, and Penicillium v.c. was the most resistant. [Kartsev V., Shikhaliev Kh. S., Geronikaki A., Medvedeva S. M., Ledenyova I. V., Krysin M. Yu., Petrou A., Ciric A., Glamoclija J., Sokovic M. / Appendix A. dithioloquinolinethiones as new potential multitargeted antibacterial and antifungal agents: Synthesis, biological evaluation and molecular docking studies // European Journal of Medicinal Chemistry. - 2019; 175; 201-214].

Various methods are known for the preparation of derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones based on substitution of the exo-sulfur atom or N-acylation. One of the ways to obtain the desired compounds is the synthesis of N-acyl derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones based on the interaction of dithioloquinolinethiones with carbonyl chlorides, including chloroacetyl chloride, in toluene with the possibility of subsequent interaction with succinimide or some other heterocyclic mercaptans to form derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones [Kartsev V., Shikhaliev Kh. S., Geronikaki A., Medvedeva S. M, Ledenyova I. V., Krysin M. Yu., Petrou A., Ciric A., Glamoclija J., Sokovic M. / Appendix A. dithioloquinolinethiones as new potential multitargeted antibacterial and antifungal agents: Synthesis, biological evaluation and molecular docking studies // European Journal of Medicinal Chemistry. - 2019; 175; 201-214]. The disadvantages of this method include the use of absolute toluene and dioxane as a medium for these interactions. The replacement of the solvent and the addition of catalytic amounts of potassium iodide can reduce the reaction time and increase the yields of the target products, as was achieved in this invention.

Closest to the claimed method is a production method based on the interaction of a salt of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones (methylthiodithiolium) with primary amines, such as aniline, ethylamine, cyclohexylamine, p-phenethidine by boiling in ethanol for about 6 hours with the formation of 1,2-dithiol-3-imines [Patent GB 1174830 (A), 1969]. According to the invention, the time of carrying out this stage of the process is reduced to 2-3 hours when using butanol as a solvent and carrying out the reaction in the presence of triethylamine. A wider range of derivatives is obtained by further modification of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones (methylthiodithiolium) salts.

The technical problem solved by the claimed invention is the search for compounds for the treatment of diseases caused by beta-coronavirus, including SARS-CoV-2, which caused the COVID-19 pandemic, which are highly effective due to the suppression of coronavirus replication and have low toxicity.

Disclosure of invention

низкой цитотоксичностью и высоким значением индекса селективности SI > 400. Данное соединение обладает низкой токсичностью, о чем свидетельствует отсутствие активности на клеточной линии здоровых клеток легочного эпителия человека, подтвержденное примерами.The technical result of the invention is a new use of a derivative of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione to suppress the replication of beta-coronaviruses, including SARS-CoV-2, characterized by high efficiency suppression of coronavirus replication in Vero E6 cell culture at low effective concentrations of EC ₅₀ (concentration of a compound that inhibits virus replication by 50% or more compared to control)

low cytotoxicity and high selectivity index SI > 400. This compound has low toxicity, as evidenced by the lack of activity on the cell line of healthy cells of the human lung epithelium, confirmed by examples.

The technical result is achieved by using a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline of general formula I to suppress the replication of beta-coronavirus, including SARS-CoV-2:

where R1, R2, R3 are a substituent selected from hydrogen; alkyl (C1-C10) straight or branched structure; alkoxy groups (C1-C10) of a straight or branched structure; aryl without substituents or substituted at positions 2,3,4,5,6; hydroxyl; amino groups; straight or branched C1-C10 alkylamino groups; nitro groups; carboxyl group; alkylcarboxylic group C1-C10, straight or branched structure; unsubstituted or substituted at the 2,3,4,5,6 positions of the arylcarboxy group; a hetarylcarboxylic group, where hetaryl is nitrogen and/or sulfur and/or an oxygen-containing five- or six-membered heterocycle,

R4 is a substituent selected from hydrogen, C1-C10 straight or branched alkyl; unsubstituted or substituted at positions 2,3,4,5,6 aryl-CH ₂ -;

or R4 is a COR5 substituent, where R5 is straight, branched or cyclic C1-C10 alkyl without substituents or containing substituents; substituted at positions 2,3,4,5,6 or unsubstituted aryl; nitrogen and/or sulfur and/or oxygen-containing five- or six-membered heterocycle; substituted at positions 2,3,4,5,6 or unsubstituted arylamine;

or R4 is a COCH ₂ R6 substituent, where R6 is a 2,3,4,5,6 substituted or unsubstituted aryloxy group; a straight-chain, branched or cyclic C1-C10 alkylamino group; a 2,3,4,5,6-substituted or unsubstituted arylamino group; hetarylamino, where hetaryl is a nitrogen and/or sulfur and/or an oxygen-containing five- or six-membered heterocycle;

or R4 is a COCH2SR7 substituent, where R7 is straight, branched or cyclic C1-C10 alkyl without substituents or containing substituents; aryl without substituents or substituted at positions 2,3,4,5,6; nitrogen and/or sulfur and/or oxygen-containing five- or six-membered heterocycle;

X is a substituent selected from sulfur, unsubstituted or substituted at the 2,3,4,5,6 positions of an arylimine.

Preferably, R1 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R2 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R3 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R4 is a substituent selected from methyl-, benzyl-, 3-phenylacryloyl-, 3-(4-methoxyphenyl)acryloyl-, 3-phenylpropanoyl-, phenylethyl, 2-(4-fluorophenoxy)acetyl-, 4-chlorophenoxyacetyl-, 2-(2,4-dichlorophenoxy)acetyl-, 4-nitrobenzoyl-, 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl-) groups; 3,4-dichlorophenylcarbamoyl-, furan-2-carbonyl-, ethylcarbamoyl-, 2-naphthylyloxyacetyl-, 2-(1,3-dioxoisoindolin-2-yl)acetyl-, 2-(1,3-dioxohexahydro-1H-isoindole -2(3H)-yl)acetyl-, 2-morpholinoacetyl-, piperidin-1-ylacetyl-, 2-(4-carbamoylpiperidin-1-yl)acetyl-, 2-(2,5-dioxopyrrolidin-1-yl) acetyl-,2-phenoxyacetyl-,2-1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)acetyl-, 2-benzo[d]oxazol-2-ylthio)acetyl-, 2 -(benzo[d]thiazol-2-yloxy)acetyl-, benzoxazol-2-ylmercaptoacetyl-, furanoyl-, 2-naphlinyloxymethylcarbonyl, 3,4-dichlorophenylcarbamoyl;

X is a substituent selected from 4-(4-chlorobenzyloxy)phenylimino-, 4-isopropylphenylimino-, 4-(6-methylbenzo[d]thiazol-2-yl)phenylimino-, 4-(N-benzoylsulfamoyl)phenylimino-, 4-ethoxyphenylimino-, 4-ethoxycarbonylphenylimino-, phenylimino-, 4-chlorophenylimino-, 3-chlorophenylimino-, 4-sulfamoylphenylimino-, 4-(N-acetylsulfamoyl)phenylimino-, 4-(N-(thiazol-2-yl) sulfamoyl)phenylimino-, 2-nicotinoylhydrazono-, 4-((4-chlorobenzyl)oxy)phenylimino-, 2-methoxyphenylimino-, 4-(N-(pyrimidin-2-yl)sulfamoyl)phenylimino-, 4-(N- (4,6-dimethylpyrimidin-2-yl)sulfamoyl)phenylimino-.

It is preferable to use a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline of general formula I for the treatment of diseases caused by beta-coronaviruses, including SARS-CoV-2, and for use in the complex therapy of diseases caused by beta-coronaviruses.

For the treatment of diseases caused by beta-coronaviruses, a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline of the general formula I is used to manufacture a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) and pharmaceutically acceptable additives. In this case, the pharmaceutical composition can be presented in the form of a single dosage form or in the form of two or more separate finished pharmaceutical forms for sequential or simultaneous administration.

Also, the compound of formula I can be used both in pure form and as an active component of new dosage forms against coronaviruses.

Compounds of general formula I for the claimed use are obtained by modification of a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline by preliminary alkylation with methyl iodide, followed by replacement of sulfur by the action of arylamines and/or by alkylation or acylation at the nitrogen atom. In this case, the synthesis of compounds I at X=S is carried out by the reaction of N-acylation with acid chlorides or anhydrides of carboxylic acids of the starting 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones. As the acid chloride, furan-2-carbonyl chloride, 2-(naphthalen-2-yloxy)acetyl chloride, 3-phenylpropanoyl chloride, 2-(4-chlorophenoxy)acetyl chloride are used, and the reaction of N-acylation with carboxylic acid chlorides is carried out in a mixture of toluene and pyridine, followed by removing the pyridinium salt and isolating the reaction product by crystallization from the filtrate. To prepare a mixture of toluene and pyridine, anhydrous toluene and pyridine are used, taken in a volume ratio for 10 parts of anhydrous toluene, from 0.8 to 1.2 parts of pyridine are taken. The reaction of N-acylation with carboxylic acid anhydrides is carried out in absolute dioxane, followed by isolation of the reaction product and its recrystallization from toluene, while using 2-chloroacetic acid anhydride in the N-acylation reaction, further alkylation of mercaptans is carried out as a result of their interaction with the resulting chloroacetamide 4,4 -dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione in absolute acetonitrile with the addition of 1.8 to 2.2 equimoles of potassium carbonate, and catalytic amounts of iodide potassium, followed by isolation of the reaction product and its recrystallization from toluene. When using 2-chloroacetic acid anhydride in the N-acylation reaction, alkylation of heterocyclic amines is carried out as a result of their interaction with the obtained chloroacetamide 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c ]quinoline-1-thione in absolute acetonitrile with the addition of catalytic amounts of potassium iodide, followed by isolation of the reaction product and its recrystallization from toluene.

The preparation of the compound of formula I at R4=H is carried out by the synthesis of imines 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones through the formation of a salt of methylthiodithiolium due to the interaction of 4,4-dimethyl -4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione with methyl iodide followed by sulfur replacement by reaction with primary amines in the presence of an excess of triethylamine (1.8 - 2 ,2 equimol), mainly 2 equimol, by boiling in absolute butanol, followed by isolation of the desired reaction product and its recrystallization from isopropyl alcohol.

The compound of formula I at X=S is obtained by the synthesis of derivatives of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones through successive substitution and N-acylation.

The method for obtaining the compound of formula (I) consists in the interaction of the salt of 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones (methylthiodithiolium) with primary amines in butanol medium, in the presence of triethylamine , makes it possible to reduce the process time to 2-3 hours and significantly expand the range of amines used, as well as to carry out further modification to obtain new derivatives with improved biological action.

For the treatment of diseases caused by beta-coronavirus, including SARS-CoV-2, the tricyclic sulfur-containing derivative of 1,2-dihydroquinoline of general formula I is administered as a pharmaceutical composition in a therapeutically effective amount.

The invention expands the arsenal of agents for the treatment and prevention of diseases caused by coronaviruses.

According to the present invention, a 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione derivative can be used to suppress the replication of betacoronavirus, including SARS-CoV-2, and therefore to treatment of diseases caused by betacoronaviruses.

Brief description of the drawings

Figure 1 shows a graph of the effectiveness of inhibition of replication of the coronavirus SARS-CoV-2 in cell culture Vero E6 and cytotoxicity for the compound 2-(naphtalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo -1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)ethanone (Id) and reference drug Remdesivir. The EC ₅₀ values are 0.51 μM and 3.76 μM for the compound 2-(naphthalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo [3,4-c]quinolin-5(4H)-yl)ethanone (Id) and reference drug Remdesivir, respectively.

Implementation of the invention

The following are definitions of terms that are used in the description of the present invention.

"Drug (drug)" - a substance (or a mixture of substances in the form of a pharmaceutical composition), in the form of tablets, capsules, injections, ointments and other ready-made forms, intended to restore, correct or change physiological functions in humans and animals, as well as to treat and disease prevention, diagnostics, anesthesia, contraception, cosmetology and other.

"Pharmaceutical composition" means a composition comprising a compound of formula 1 and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, adjuvants, delivery vehicles, preservatives, stabilizers, bulking agents, grinders, humectants, emulsifiers, suspending agents, thickeners, sweeteners, fragrances, flavors. Prolonged action of the composition can be provided with agents that slow down the absorption of the active principle, for example, aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols and mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of fillers are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and spreading agents are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. The pharmaceutical composition for oral, sublingual, intramuscular, intravenous, subcutaneous, alone or in combination with another active principle, can be administered to animals and humans in a standard form of administration, as a mixture with conventional pharmaceutical carriers. Suitable unit administration forms include oral forms such as tablets, gelatin capsules, pills, powders, granules, oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, topical, transdermal, subcutaneous, intramuscular, intravenous, intranasal administration forms.

The term “therapeutically effective amount” means an amount of an active ingredient that (1) treats or prevents a particular disease, condition, or disorder, (2) relieves, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (3) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein. The term "therapeutically effective amount" means that amount of a compound of formula (I) which is sufficient to provide the desired therapeutic effect. In this case, the daily dose in adults is usually 1.0 ~ 500 mg, preferably 10 ~ 300 mg. Therefore, during the preparation of the pharmaceutical composition of the drug of the present invention in the form of dosage units, it is necessary to take into account the above effective dosage, and each unit dosage of the drug should contain 10 ~ 500 mg of the compound of general formula I, preferably 50 ~ 300 mg. In a preferred embodiment, the therapeutically effective amount is from 0.01 to 1 mg/kg of the subject's body weight. According to the instructions of the doctor or pharmacist, these drugs can be taken several times over a certain period of time (preferably from one to six times). At the same time, the dosage of an agent containing a compound of general formula (I) in patients can be adjusted depending on the therapeutic efficacy and bioavailability of the active ingredient in the body, the rate of their metabolism and excretion from the body, as well as depending on the age, sex and stage of the patient's disease. .

Drugs may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or topically).

EC ₅₀ , or half-maximal inhibition concentration, is a measure of the effectiveness of a ligand in an inhibitory biochemical or biological interaction. EC ₅₀ is a quantitative indicator that shows how much inhibitor ligand is needed to inhibit a biological process by 50%, including the suppression of viral replication in cell culture.

IC ₅₀ is the concentration of half-maximal inhibition of the target protein in experiments with a fluorogenic substrate in the protein-substrate-inhibitor test system.

TC ₅₀ or CC ₅₀ - the concentration of the drug, at which 50% death of the studied cell culture is observed.

SI - selectivity index, the ratio of compound toxicity and inhibitory activity against betacoronavirus (TC50 / IC50).

Catalytic amounts of catalyst - the minimum amount of a substance that ensures the reaction, taking into account various technological factors for the reaction, primarily the dependence of the temperature / duration of the reaction (the more catalyst, the faster the reaction, the higher the temperature, the acceleration of the reaction, the catalyst can be taken less).

The biological activity of the compound of formula (I), which consists in inhibiting the reproduction of beta-coronavirus, was previously identified using the method of molecular modeling. Based on docking, derivatives of 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thiones with better parameters were identified, which subsequently found experimental confirmation of their inhibitory activity . Molecular docking was implemented using the M.V. Lomonosov of the SOL program [Romanov A.N., Kondakova O.A., Grigoriev F.V., Sulimov A.V., Lushchekina S.V., Martynov Ya.B., Sulimov V.B. / Computer-aided drug development: SOL docking program // Computational methods and programming. - 2008; 9(2); 64-84; Sulimov VB, Ilin IS, Kutov DC, Sulimov AV / Development of docking programs for Lomonosov supercomputer // Journal of the Turkish Chemical Society Section A: Chemistry. - 2020; 7(1); 259-276], which determines the protein-ligand binding energy, and has received international recognition among similar software products [Alexey V. Sulimov, Danil C Kutov, Igor V. Oferkin, Ekaterina V. Katkova, and Vladimir B. Sulimov / Application of the Docking Program SOL for CSAR Benchmark // J. Chem. inf. Model. - 2013; 53; 1946-1956]. The identified group of compounds was then additionally tested using the PM7 quantum-chemical semi-empirical method [Sulimov AV, Kutov DC, Taschilova AS, Ilin IS, Stolpovskaya NV, Shikhaliev Kh S., Sulimov VB / In search of non-covalent inhibitors of SARS-CoV -2 main protease: Computer aided drug design using docking and quantum chemistry // Supercomputing Frontiers and Innovations. - 2020; 7(3); 41-56] with a continuum solvent model: additional local optimization from the position of the ligand found during docking in the protein and calculation of the protein-ligand binding enthalpy. For the experiment, we selected compounds whose molecules for this target had a sufficiently negative scoring and a sufficiently negative protein–ligand binding enthalpy. It is among these compounds, selected on the basis of their high binding energy to the main protease of the SARS-CoV-2 coronavirus, that compounds were found that effectively suppress the replication of this coronavirus in Vero E6 cell culture. A very close analogue of the compound 2-(naphthalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinoline-5( 4H)-yl)ethanone (Id), namely the compound 2-(naphthalen-2-yloxy)-1 -(4,4,7-trimethyl-1-thioxo-1H-[1,2]dithiolo -c]quinolin-5(4H)-yl)ethanone (Ic), not only inhibits SARS-CoV-2 coronavirus replication in Vero E6 cell culture with an effective concentration of EC ₅₀ = 1.16 μM, has a high selectivity index SI > 86, but and inhibits the main protease of the SARS-CoV-2 coronavirus with an effective inhibitory concentration of IC ₅₀ = 2.74 μM.

The selected compounds effectively suppress the replication of betacoronaviruses in cell cultures.

The following is a more detailed description of embodiments of the present invention with the achievement of the claimed result. The following examples illustrate but do not limit the present invention.

All reagents used are commercially available, the progress of the reaction was monitored by thin layer chromatography (TLC), and the reaction time is for illustration only; the structure and purity of all isolated compounds were confirmed by at least one of the following methods: TLC (TLC plates precoated with Merck silica gel 60 F ₂₅₄ ), mass spectrometry or nuclear magnetic resonance (NMR). Product yield is for illustration purposes only. ¹ H NMR spectra were recorded on an Agilent MR 400+ spectrometer (at operating frequencies of 400 and 100 MHz, respectively) under normal conditions in DMSO-D6 solutions unless otherwise noted, with tetramethylsilane (TMS) as an internal standard, ppm (m .d.). Chromatographic analysis was carried out on an Agilent Technologies 1260 infinity instrument with an Agilent 6230 TOF LC/MS mass detector (high-resolution time-of-flight mass detector), ionization method - double electrospray (dual-ESI). Recording and registration of signals was carried out in positive polarity; nebulizer (N ₂ ) 20 psig, drying gas (N ₂ ) 6 ml/min, 325°C; mass detection range is 50-2000 Dalton. Capillary voltage 4.0 kV, fragmentator +191 V, skimmer +66 V, OctRF 750 V. Chromatography conditions: Poroshell 120 EC-C18 column (4.6 × 50 mm; 2.7 µm). Gradient elution: acetonitrile/water (0.1% formic acid); flow rate 0.4 ml/min. Research results processing software - MassHunter Workstation / Data Acquisition V.06.00. Melting points were determined on a Stuart SMP30 apparatus.

All procedures, unless otherwise noted, were performed at room or ambient temperature, i.e. in the range of 20-25°C.

Drying of the products to constant weight was carried out at a temperature of 35-45°C at atmospheric pressure or using a vacuum drying Cabinet at a residual pressure of 0.35±0.005 kg/cm ² (35±5 kPa).

Distilled water was used to wash the precipitates/filtrate unless otherwise noted.

Target sulfur-containing derivatives of 1,2-dihydroquinoline I were obtained by a multistage synthesis, including the preparation of the starting dihydroquinolines followed by their heterocyclization with molecular sulfur to form 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1 -thions and further carrying out the reactions of their structural modification in three ways (A, B, C), presented in Scheme 1.

Synthesis of the original 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones is carried out in two stages from the corresponding anilines according to known methods [Brown, J. P. J. Chem. soc. C, 1968, 1074-1075; Reddelin C, Thurm A. // Ber., 1932, 65, 1511-1521; Shikhaliev, X.S., Shmyreva Zh.V., Zalukaev L.P. / Acylation of 4,5-dihydro-4,4-dimethyl-5H-2,3-dithiolo[5,4-c]quinoline-1-thione // Zh. organ, chemistry. - 1988; 24(1); 232-233; Shikhaliev Kh.S., Pigarev V.V., Soloviev A.S., Shatalov G.V. / Synthesis of N-(2-hetarylacetyl)-8-R-4,5-dihydro-4,4-dimethyl-2,3-dithiolo[5,4-c]quinoline-1-thiones // Izv. universities. Chemistry and chem. technology. - 2000; 43(2); .93-94; Medvedeva S.M.; Leshcheva E.V.; Shikhaliev Kh.S. Solov'ev A.S. / Novel heterocyclic systems based on 8-R-4,5-dihydro-4,4-dimethyl[1,2]dithiolo[3,4-c]quinoline-l-thiones // Chem Heterocycl Compd. - 2006; 42; 534-539; Kartsev V., Shikhaliev Kh. S., Geronikaki A., Medvedeva S. M., Ledenyova I. V., Krysin M. Yu., Petrou A., Ciric A., Glamoclija J., Sokovic M. / Appendix A. dithioloquinolinethiones as new potential multitargeted antibacterial and antifungal agents: Synthesis, biological evaluation and molecular docking studies // European Journal of Medicinal Chemistry. - 2019; 175; 201-214]. When performing these stages of synthesis (obtaining 2,2,4-trimethyl-1,2-dihydroquinolines and 4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thiones), there may be any methods and means known from the prior art are used.

Route A: preparation of a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline based on N-acylation reactions with acid chlorides or anhydrides of carboxylic acids with the formation of final compounds I at X=S.

Stage 1: prepare a mixture of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione containing substituents on the benzene ring, including alkyl groups ( C1-C10) unbranched or branched structure; alkoxy groups (C1-C10) of a straight or branched structure; aryls without substituents or substituted at positions 2,3,4,5,6; a hydroxyl group; an amino group; alkylamino group, where alkyl C1-C10 straight or branched structure; a nitro group; a carboxyl group; alkylcarboxyl group, where alkyl C1-C10 straight or branched structure; an arylcarboxy group, where aryl is unsubstituted or substituted at positions 2,3,4,5,6; a hetarylcarboxylic group, where hetaryl is nitrogen and/or sulfur and/or an oxygen-containing five- or six-membered heterocycle and the corresponding carboxylic acid chloride, taken from the calculation that per 0.01 mol of 4,4-dimethyl-4,5-dihydro-1H- [1,2]dithiolo[3,4-c]quinoline-1-thione take 0.012-0.013 mol of carboxylic acid halide, in a mixture of absolute toluene and pyridine, taken in a volume ratio of 10:1 in an amount of 70-100 ml. Boil for 3-5 hours, filter off the resulting salt of pyridinium chloride, the filtrate is evaporated, the precipitate is filtered off and recrystallized from toluene. The yield of the obtained product is 70-85%.

As a carboxylic acid halide, acid chlorides and acid bromides of aliphatic acids containing branched and unbranched hydrocarbon radicals, as well as cyclic hydrocarbon radicals, unsaturated hydrocarbon radicals, including acid chloride of acetic, trifluoroacetic, propanoic, butanoic, isobutanoic, pentanoic, 2 -methylbutanoic, 3-methylbutanoic, hexanoic, propenoic, buteneic, cinnamic, etc. acids, acid chlorides and bromides of aromatic carboxylic acids, including benzoic, 2-chlorobenzoic, 3-chlorobenzoic, 4-chlorobenzoic, 2,4-dichlorobenzoic, 2-bromobenzoic, 3-bromobenzoic, 4-bromobenzoic, 4-iodobenzoic, 2- iodobenzoic, 3-iodobenzoic, 2-fluorobenzoic, 3-fluorobenzoic, 4-fluorobenzoic, 2-methylbenzoic, 3-methylbenzoic, 4-methylbenzoic, 2,4-dimethylbenzoic, 3,4-dimethylbenzoic, 2,5-dimethylbenzoic, 2- methoxybenzoic, 3-methoxybenzoic, 4-methoxybenzoic, 4-nitrobenzoic, naphthalene-2-carboxylic, naphthalene-3-carboxylic, etc. acids, acid chlorides and bromides of heterocyclic carboxylic acids, including furan-2-carboxylic, thiophene-2-carboxylic, etc. acids.

Or a mixture of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and the corresponding carboxylic acid anhydride, including aliphatic acid anhydrides containing hydrocarbon branched and straight chain radicals, as well as cyclic hydrocarbon radicals, including 2-chloroacetic acid anhydride, taken per 0.01 mol of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3 ,4-c]quinoline-1-thione take 0.012 - 0.013 mol of carboxylic anhydride, boil in 70-100 ml of absolute dioxane for 3-6 hours, the precipitate is filtered off, washed with water, dried and recrystallized from toluene. The yield of the obtained product is 60-85%.

If 2-chloroacetic acid anhydride acts as an acylating agent, further modification is possible (stage 2): alkylation of various mercaptans and heterocyclic amines with 4,4-dimethyl-4,5-dihydro-1H chloroacetamide obtained at the first stage of route A -[1,2]dithiolo[3,4-c]quinoline-1-thione.

Stage 2: This stage consists in the alkylation of various heterocyclic amines or mercaptans with chloroacetamide 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione obtained in the first stages.

Benzo[d]oxazol-2-ylthiol, benzo[d]thiazol-2-ylthiol, pyridin-2-ylthiol, pyrimidin-2-ylthiol, imidazol-2-ylthiol, 1,2,4- triazol-2-ylthiol, 1,3,4-thiazol-2-ylthiol, etc.

As a heterocyclic amine, morpholine, piperidine, piperidine-4-carboxamide, 4-methylpiperidine, pyrrolidine, 4-methylpiperazine, 3-methylpiperidine-, ethyl piperidine-4-carboxylate, 4-phenylpiperazine, 4-(2-chlorophenyl) can be used. piperazine, 4-(4-chlorophenyl)piperazine, 4-(4-methylphenyl)piperazine, etc.

Prepare a mixture of chloroacetamide 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and the corresponding mercaptan, taken from the calculation that per 0.01 mol of chloroacetamide 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione take at least 0.01 mol of mercaptan, the resulting mixture is dissolved in 70-100 ml of absolute acetonitrile, add at least 0.02 mol of potassium carbonate and 0.001-0.0015 mol of potassium iodide, boil for 3-5 hours, cool, filter, the precipitate is recrystallized from toluene. The yield of the obtained product is 70-80%.

Or, a mixture of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and the corresponding heterocyclic amine is prepared, taken from the calculation that by 0.01 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione mole of chloroacetamide take 0.022-0.023 mole of heterocyclic amine, the resulting mixture is dissolved in 70-100 ml of absolute acetonitrile, add 0.001 - 0.0015 mol of potassium iodide, boil for 3-5 hours, cool, filter, the precipitate is recrystallized from toluene. The yield of the obtained product is 50-70%.

Pathway B: preparation of a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline as a result of sulfur substitution in the reaction of a methylthiodithiolium salt with primary amines and the formation of imines 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4 -c]quinoline-1-thione I at R ₄ =H.

A solution of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione is prepared in acetone (0.01 mol in 50-100 ml), iodide is added methyl, taken from the calculation that at least 0.015 mol of methyl iodide , boil for 3-5 hours, cool, filter off the methylthiodithiolium salt. A mixture of the resulting salt of methylthiodithiolium, the corresponding primary amine and triethylamine is prepared, taken on the basis that 0.011 - 0.012 mol of primary amine and at least 0.02 mol of triethylamine are taken per 0.01 mol of methylthiodithiolium salt, the resulting mixture is boiled in absolute butanol for 2-3 hours, cooled, the solvent is distilled off under reduced pressure, the precipitate is filtered off, recrystallized from isopropyl alcohol. The yield of reaction products is 60-80%.

As primary amines, aniline, 2-methylphenylamine, 3-methylphenylamine, 4-methylphenylamine, 2,4-dimethylphenylamine, 3,4-dimethylphenylamine, 2-methoxyphenylamine, 3-methoxyphenylamine, 4-methoxyphenylamine, 2-ethoxyphenylamine, 3 -ethoxyphenylamine, 4-ethoxyphenylamine, 2-chlorophenylamine, 3-chlorophenylamine, 4-chlorophenylamine, 2-bromophenylamine, 3-bromophenylamine, 4-bromophenylamine, 2-nitrophenylamine, 3-nitrophenylamine, 4-nitrophenylamine, 4-phenoxymethylphenylamine, 4-( 4-chlorophenoxymethyl)-phenylamine,] N[-acetyl-4-aminobenzenesulfamide, N-pyrimidin-2-yl-4-aminobenzenesulfamide, etc.

Pathway C (B+A): successive sulfur substitution (stage 1) and N-acylation or N-carbamoylation (stage 2) with the formation of final compounds I.

Step 1: prepare a solution of 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione in acetone (0.01 mol in 50-100 ml) , methyl iodide is added, taken on the basis that at least 0.015 mol of methyl iodide, and boil the resulting mixture for 3-5 hours, cool, filter off the methylthiodithiolium salt. A mixture of the resulting methylthiodithiolium salt, the corresponding primary amine and triethylamine is prepared, taken on the basis that 0.011-0.012 mol of primary amine and at least 0.02 mol of triethylamine are taken per 0.01 mol of methylthiodithiolium salt, the resulting mixture is boiled in absolute butanol for 2-3 hours, cooled, filtered off, the solvent is distilled off under reduced pressure, the precipitate is filtered off, recrystallized from isopropyl alcohol. The yield of reaction products is 60-80%.

As primary amines, aniline, 2-methylphenylamine, 3-methylphenylamine, 4-methylphenylamine, 2,4-dimethylphenylamine, 3,4-dimethylphenylamine, 2-methoxyphenylamine, 3-methoxyphenylamine, 4-methoxyphenylamine, 2-ethoxyphenylamine, 3 -ethoxyphenylamine, 4-ethoxyphenylamine, 2-chlorophenylamine, 3-chlorophenylamine, 4-chlorophenylamine, 2-bromophenylamine, 3-bromophenylamine, 4-bromophenylamine, 2-nitrophenylamine, 3-nitrophenylamine, 4-nitrophenylamine, 4-phenoxymethylphenylamine, 4-( 4-chlorophenoxymethyl)-phenylamine, N-acetyl-4-aminobenzenesulfamide, N-pyrimidin-2-yl-4-aminobenzenesulfamide, etc.

Stage 2. This stage consists in the acylation or carbamoylation of the imine 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione, obtained in the first stage with carboxylic acid chloride acid or isocyanate, respectively.

As a carboxylic acid halide, acid chlorides and acid bromides of aliphatic acids containing branched and unbranched hydrocarbon radicals, as well as cyclic hydrocarbon radicals, unsaturated hydrocarbon radicals, including acid chloride of acetic, trifluoroacetic, propanoic, butanoic, isobutanoic, pentanoic, 2 -methylbutanoic, 3-methylbutanoic, hexanoic, propenoic, buteneic, cinnamic, etc. acids, acid chlorides and bromides of aromatic carboxylic acids, including benzoic, 2-chlorobenzoic, 3-chlorobenzoic, 4-chlorobenzoic, 2,4-dichlorobenzoic, 2-bromobenzoic, 3-bromobenzoic, 4-bromobenzoic, 4-iodobenzoic, 2- iodobenzoic, 3-iodobenzoic, 2-fluorobenzoic, 3-fluorobenzoic, 4-fluorobenzoic, 2-methylbenzoic, 3-methylbenzoic, 4-methylbenzoic, 2,4-dimethylbenzoic, 3,4-dimethylbenzoic, 2,5-dimethylbenzoic, 2- methoxybenzoic, 3-methoxybenzoic, 4-methoxybenzoic, 4-nitrobenzoic, naphthalene-2-carboxylic, naphthalene-3-carboxylic, etc. acids, acid chlorides and bromides of heterocyclic carboxylic acids, including furan-2-carboxylic, thiophene-2-carboxylic, etc. acid.

As an isocyanate, phenylisocyanate, 2-methylphenylisocyanate, 3-methyl phenylisocyanate, 4-methylphenylisocyanate, 2-ethylphenylisocyanate, 3-ethylphenylisocyanate, 4-ethylphenylisocyanate, 2-methoxyphenylisocyanate, 3-methoxyphenylisocyanate, 4-methoxyphenylisocyanate, 2-chlorophenylisocyanate, 3 -chlorophenylisocyanate, 4-chlorophenylisocyanate, 3,4-dichlorophenylisocyanate, 2-bromophenylisocyanate, 3-bromophenylisocyanate, 4-bromophenylisocyanate, naphthalene-2-ylisocyanate, naphthalene-1-ylisocyanate, etc.

A mixture of the imine 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione obtained at the 1st stage and the corresponding carboxylic acid chloride, taken from the calculation that per 0.01 mol of imine 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione take 0.012-0.013 mol of the corresponding carboxylic acid chloride, boil in a mixture of absolute toluene and pyridine, taken in a volume ratio of 10:1 in an amount of 70-100 ml, for 3-5 hours, the pyridinium chloride salt is filtered off, the filtrate is evaporated, the precipitate is filtered off and recrystallized from toluene. The outputs of the target products are 70-85%.

Or, a mixture of the imine 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione obtained at the 1st stage and the corresponding isocyanate, taken on the basis that 0 01 mol of imine 4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione take 0.011-0.012 mol of isocyanate, the resulting mixture is boiled at 70-100 ml of absolute dioxane for 5-6 hours, the precipitate is filtered off, washed with water, after drying, recrystallized from toluene. Yield 60-85%.

Methods for the synthesis of final compounds I, methods for their preparation, as well as spectral data, physico-chemical characteristics are presented in the examples below.

Derivatives of the tricyclic sulfur-containing derivative of quinolin-2(1H)-one I were obtained by the above methods for the first time.

EXAMPLE 1

Synthesis of 2-(naphthalen-2-yloxy)-1-(4,4,8-trimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl) ethanone (Ia) was carried out via route A.

A mixture of 0.01 mol 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinolin-1-thione and 0.012 mol 2-(naphthalen-2-yloxy )acetyl chloride was boiled in 100 ml of a mixture of anhydrous toluene and pyridine (10:1) for 5 hours, the pyridinium salt was filtered off, the filtrate was evaporated, the precipitate that formed was filtered off and recrystallized from toluene. Yield 80%.

Orange powder, melting point 153-154°C; 1.65 (br. s, 6H, (CH3) ₂ ), 2.35 ( _s , _3H , CH3), 4.79 (br. _s , 2H, CH2CO), 6.77 (s, 1H, H-Ar), 6.81 (d, J=8.78 Hz, 1H, H-6(7)), 7.22 (d, J=8.78 Hz, 1H, H-7(6)), 7.28 (t, J=8.04 Hz, 1H, H- Ar), 7.38 (t, J=8.04 Hz, 1H, H-Ar), 7.47 (d, J=8.04 Hz, 1H, H-Ar), 7.58 (d, J=8.04 Hz, 1H, H-Ar) , 7.66 (d, J=8.04 Hz, 1H, H-Ar), 7.75 (d, J=8.04 Hz, 1H, H-Ar), 8.98 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₅ H ₂₁ NO ₂ S ₃ +H ⁺ , 464.0808; found, 464.0811.

EXAMPLE 2

Synthesis of 1-(8-methoxy-4,4-dimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl1)-2-(naphthalene-2- yloxy)ethanone (Ib).

The synthesis was carried out analogously to example 1, while 8-methoxy-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 2-(naphthalen-2-yloxy)acetyl chloride. Yield 83%.

Orange powder, melting point 136-137°C; 1.33 (br. s, 3H, (CH3) ₂ ), 2.04 (br. s, _3H , (CH3) ₂ ), 3.79 ( _s , _3H , CH3O), 4.78 (br. s, 2H, CH ₂ CO), 6.74 (s, 1H, H-Ar), 6.82 (d, J=8.75 Hz, 1H, H-6(7)), 7.00 (d, J=8.75 Hz, 1H, H-7(6 )), 7.30 (t, J=8.23 Hz, 1H, H-Ar), 7.38 (t, J=8.23 Hz, 1H, H-Ar), 7.55 (d, J=8.23 Hz, 1H, H-Ar) , 7.58 (d, J=8.23 Hz, 1H, H-Ar), 7.67 (d, J=8.23 Hz, 1H, H-Ar), 7.75 (d, J=8.23 Hz, 1H, H-Ar), 9.03 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₅ H ₂₁ NO ₃ S ₃ +H ⁺ , 480.0758; found, 480.0755.

EXAMPLE 3

Synthesis of 2-(naphthalen-2-yloxy)-1-(4,4,7-trimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl) ethanone (Ic)

The synthesis was carried out analogously to example 1, while 4,4,7-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 2- (naphthalen-2-yloxy)acetyl chloride. Yield 77%.

Orange powder, melting point 144 - 145°C; 1.65 (br. s, 6H, (CH ₃ ) ₂ ), 2.32 (s, 3H, CH ₃ ), 4.84 (br. s, 2H, CH ₂ CO), 6.77 (s, 1H, H-6), 6.79 (d, J=8.15 Hz, 1H, H-Ar), 7.21 (d, J=8.35 Hz, 1H, H-8), 7.30 (t, J=8.15 Hz, 1H, H-Ar), 7.36 (s , 1H, H-Ar), 7.39 (t, J=8.15 Hz, 1H, H-Ar), 7.56 (d, J=8.15 Hz. 1H, H-Ar), 7.62 (d, J=8.15 Hz, 1H , H-Ar), 7.75 (d, J=8.15 Hz, 1H, H-Ar), 9.03 (d, J=8.35 Hz, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₅ H ₂₁ NO ₂ S ₃ +H ⁺ , 464.0808; found, 464.0812.

EXAMPLE 4

Synthesis of 2-(naphthalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinoline-5(4H)- yl)ethanone (Id)

The synthesis was carried out analogously to example 1, while 4,4,7,8-tetramethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 2-(naphthalen-2-yloxy)acetyl chloride. Yield 80%.

Orange powder, melting point 189-190°C; 1.64 (br. s, 6H, (CH ₃ ) ₂ ), 2.24 (s, 6H, 2CH ₃ ), 4.82 (br. s, 2H, CH ₂ CO), 6.76 (s, 1H, H-6), 6.82 (d, J=8.71 Hz, 1H, H-Ar), 7.30 (d, J=8.71 Hz, 1H, H-Ar), 7.32 (s, 1H, H-Ar), 7.38 (d, J=8.71 Hz , 1Н, Н-Ar), 7.57 (d, J=8.71 Hz, 1Н, Н-Ar), 7.66 (d, J=8.71 Hz, 1H, Н-Ar), 7.75 (d, J=8.71 Hz, 1H , H-Ar), 8.93 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₆ H ₂₃ NO ₂ S ₃ +H ⁺ , 478.0965; found, 478.0964.

EXAMPLE 5

Synthesis of 3-phenyl-1-(4,4,6-trimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)propan-1-one ( Ie)

The synthesis was carried out analogously to example 1, while 4,4,6-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 3- phenylpropanoyl chloride. Yield 74%.

Orange powder, melting point 117-118°C; 1.04 (s, 3H, CH ₃ ), 2.07-2.15 (m, 1H, CH ₂ ), 2.22 (s, 3H, (CH ₃ ) ₂ ), 2.28 (s, 3H, (CH ₃ ) ₂ ), 2.32- 2.39 (m, 1H, CH ₂ ), 2.52 (d, J=7.66 Hz, 2H, CH ₂ CO), 6.84 (d, J=7.01 Hz, 2H, H-Ar), 7.04 (t, J=7.01 Hz , 1H, H-Ar), 7.12 (t, J=7.01 Hz, 2H, H-Ar), 7.32 (t, J=6.18 Hz, 1H, H-8), 7.35 (d, J=6.18 Hz, 1H , H-7), 8.91 (d, J=6.18 Hz, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₂ H ₂₁ NOS ₃ + H ⁺ , 412.0859; found, 412.0862.

EXAMPLE 6

Synthesis of 2-(4-chlorophenoxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl) ethanone (If)

The synthesis was carried out analogously to example 1, while 4,4,7,8-tetramethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 2-(4-chlorophenoxy)acetyl chloride. Yield 70%.

Orange powder, melting point 181-182°C; ¹ H NMR (DMSO-d ₆ , 500 MHz) δ ppm: 1.63 (br. s, 6H, (CH ₃ ) ₂ ), 2.22 (s, 6H, 2CH ₃ ), 4.66 (s, 2H, CH ₂ CO ), 6.54 (d, J=9.01 Hz, 2H, H-Ar), 7.13 (d, J=9.01 Hz, 2H, H-Ar), 7.32 (s, 1H, H-6), 8.82 (s, 1H , H-9); HPLC-HRMS (ESI) calculated for C ₂₂ H ₂₀ CINO ₂ S ₃ + H ⁺ , 462.0419; found, 462.0420.

EXAMPLE 7

Synthesis of 1-(8-methoxy-4,4-dimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)-2-(piperidin-1- yl)ethanone (Ig)

A mixture of 0.01 mol of 8-methoxy-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 0.012 mol of 2-chloroacetic anhydride was boiled in 100 ml of anhydrous dioxane for 6 hours, the precipitate that formed was filtered off, washed with water, dried, and recrystallized from toluene. To a mixture of 0.01 mol of 8-methoxy-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione chloroacetamide and 0.022 mol of piperidine dissolved in 100 ml of anhydrous acetonitrile, 0.001 mol of potassium iodide was added, the resulting mixture was boiled for 5 hours, cooled, the precipitate formed was filtered off, dried, and recrystallized from toluene. Yield 70%.

Orange powder, melting point 124-125°C; ¹ H NMR (DMSO-d ₆ , 500 MHz) δ ppm: 1.15-1.17 (m, 8H, 4 CH ₂ ), 1.99 (br. s, 6H, (CH ₃ ) ₂ ), 2.51 (m, 2H, CH ₂ ), 2.95 (br. s, 2H, COCH ₂ ), 3.77 (s, 3H, CH ₃ O), 6.93 (d, J=8.66 Hz, 1H, H-6(7)); 7.39 (d, J=8.66 Hz, 1H, H-7(6)), 8.72 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₀ H ₂₄ N ₂ O ₂ S ₃ + H ⁺ , 421.1074; found, 421.1074.

EXAMPLE 8

Synthesis of 1-(2-oxo-2-(4,4,8-trimethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)ethyl)piperidine -4-carboxamide (Ih)

The synthesis was carried out analogously to example 7, while 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and piperidine- 4-carboxamide. Yield 50%.

Orange powder, melting point 115-116°C; ¹ H NMR (DMSO-d ₆ , 500 MHz) δ ppm: 1.26-1.34 (m, 2H, CH ₂ ), 1.46-1.49 (m, 2H, CH ₂ ), 1.70 (br. s, 6H, (CH ₃ ) ₂ ), 1.85–1.87 (m, 3H, CH ₂ ), 2.34 (s, 3H, CH ₃ ), 2.51 (m, 1H, CH ₂ ), 3.00 (br. s, 2H, COCH ₂ ), 6.63 ( br. s, 1H, NH ₂ ), 7.02 (br. s, 1H, NH ₂ ), 7.20 (d, J=8.06 Hz, 1H, H-6(7)); 7.36 (d, J=8.06 Hz, 1H, H-7(6)), 8.90 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₁ H ₂₅ N ₃ O ₂ S ₃ + H ⁺ , 448.1183; found, 448.1186.

EXAMPLE 9

Synthesis of 2-(benzo[d]oxazol-2-ylthio)-1-(4,4,6,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinoline-5 (4H)-yl)ethanone (Ii)

A mixture of 0.01 mol of 4,4,6,8-tetramethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 0.012 mol of 2-chloroacetic anhydride, boiled in 100 ml of anhydrous dioxane for 6 hours, the precipitate was filtered off, washed with water, and after drying, recrystallized from toluene. To a mixture of 0.01 mol of 4,4,6,8-tetramethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione chloroacetamide and 0.01 mol of benzo[ d]oxazole-2-thiol in 100 ml of anhydrous acetonitrile, 0.02 mol of potassium carbonate and 0.001 mol of potassium iodide were added, boiled for 4 hours, cooled, the precipitate formed was filtered off, dried, and recrystallized from toluene. Yield 78%.

Orange powder, melting point 156-157°C; ¹ H NMR (DMSO-d ₆ , 500 MHz) δ ppm: 1.16 (s, 3H, (CH ₃ ) ₂ ), 2.23 (s, 3H, (CH ₃ ) ₂ ), 2.33 (s, 3H, CH ₃ ) , 2.40 (s, 3H, CH ₃ ), 3.79 (d, J=16.27 Hz, 1H, CH ₂ ), 4.11 (d, J=16.27 Hz, 1H, CH ₂ ), 7.24 (t, J=7.76 Hz, 1H, H-Ar), 7.26 (s, 1H, H-7); 7.28 (t, J=7.76 Hz, 1H, H-Ar), 7.52 (d, J=7.76 Hz, 1H, H-Ar), 7.54 (d, J=7.76 Hz, 1H, H-Ar), 8.80 ( s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₃ H ₂₀ N ₂ O ₂ S ₄ + H ⁺ , 485.0481; found, 485.0479.

EXAMPLE 10

Synthesis of 4-((4,4,7-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene)amino)benzenesulfamide (Ij) was carried out via route B .

To a solution of 0.01 mol of 4,4,7-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione in 100 ml of acetone was added 0.015 mol of methyl iodide and boiled for 3 hours, cooled, filtered off the methylthiodithiolium salt, 0.01 mol of which, after drying, boiled with 0.011 mol of 4-aminobenzenesulfamide and 0.02 mol of triethylamine in 100 ml of dry butanol for 3 hours, cooled, the solvent was distilled off under reduced pressure, the precipitate was filtered off, dried, and recrystallized from dioxane. Yield 76%.

Yellow powder, melting point 306-307°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.42 (br. s, 6H, (CH ₃ ) ₂ ), 2.19 (s, 3H, CH ₃ ), 6.20 (br. s, 1H, NH) , 6.47 (d, J=8.10 Hz, 1H, H-8); 6.56 (s, 1Н, Н-6), 7.18 (d, J=8.60 Hz, 2Н, Н-Ar), 7.32 (br. s, 2Н, NH ₂ ), 7.85 (d, J=8.60 Hz, 2Н, H-Ar), 8.40 (d, J=8.10 Hz, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₁₉ H ₁₉ N ₃ O ₂ S ₃ + H ⁺ , 418.0713; found, 418.0709.

EXAMPLE 11

Synthesis of N-((4-((4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene)amino)phenyl)sulfonyl) acetamide (Ik).

The synthesis was carried out analogously to example 10, but 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and N-( (4-aminophenyl)sulfonyl)acetamide. Yield 68%.

Yellow powder, melting point 288-289°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.42 (br. s, 6H, (CH ₃ ) ₂ ), 1.93 (s, 3H, CH ₃ CO), 2.19 (s, 3H, CH ₃ ) , 6.12 (br. s, 1Н, NH), 6.67 (d, J=8.06 Hz, 1Н, Н-6(7)); 6.88 (d, J=8.06 Hz, 1H, Н-7(6)), 7.24 (d, J=8.67 Hz, 2Н, Н-Ar), 7.93 (d, J=8.67 Hz, 2Н, Н-Ar) , 8.33 (s, 1H, H-9), 12.05 (br. s, 1H, CONH); HPLC-HRMS (ESI) calculated for C ₂₁ H ₂₁ N ₃ O ₃ S ₃ + H ⁺ , 460.0819; found, 460.0816.

EXAMPLE 12

Synthesis of 4-((8-ethoxy-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene)amino)-N-(pyrimidine- 2-yl)benzenesulfamide (II)

The synthesis was carried out analogously to example 10, but 8-ethoxy-4,4-dimethyl-4,5-dihydro-1 H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 4-amino-N-(pyrimidin-2-yl)benzenesulfamide. Yield 80%.

Yellow powder, melting point 272-273°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.22 (t, 3H, J=13.89 Hz, CH ₃ CH ₂ O), 1.39 (br. s, 6H, (CH ₃ ) ₂ ), 3.84 ( q, 2Н, J=13.89 Hz, CH ₂ O), 5.95 (br. s, 1Н, NH), 6.67 (d, J=8.11 Hz, 1H, Н-6(7)); 6.73 (d, J=8.11 Hz, 1H, H-7(6)), 7.05 (t, J=4.89 Hz, 1H, H-Het), 7.20 (d, J=8.60 Hz, 2H, H-Ar) , 8.01 (d, J=8.60 Hz, 2H, H-Ar), 8.16 (s, 1H, H-9). 8.51 (d, J=4.89 Hz, 2Н, H-Het), 11.79 (br. s, 1Н, SNH); HPLC-HRMS (ESI) calculated for C ₂₄ H ₂₃ N ₅ O ₃ S ₃ + H ⁺ , 526.1037; found, 526.1037.

EXAMPLE 13

Synthesis of 4-((4-chlorobenzyl)oxy)-N-(4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene) aniline (Im)

The synthesis was carried out analogously to example 10, but 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 4-( (4-chlorobenzyl)oxy)aniline. Yield 73%.

Yellow powder, melting point 142-143°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.42 (br. s, 6H, (CH ₃ ) ₂ ), 2.17 (s, 3H, CH ₃ ), 5.12 (br. s, 2H, CH ₂ O), 6.09 (br. s, 1H, NH), 6.67 (d, J=8.0 Hz, 1H, Н-6(7)) 6.90 (d, J=8.0 Hz, 1H, Н-7(6)) , 6.99 (d, J=8.68 Hz, 2H, H-Ar), 7.08 (d, J=8.68 Hz, 2H, H-Ar), 7.76 (d, J=8.38 Hz, 2H, H-Ar), 7.51 (d, J=8.38 Hz, 2H, H-Ar), 8.41 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₆ H ₂₃ ClN ₂ OS ₂ + H ⁺ , 479.1014; found, 479,1016.

EXAMPLE 14

Synthesis of N-((4-((4,4,5,8-tetramethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-ylidene)amino)phenyl) sulfonyl)acetamide (In). To obtain the In compound, the following is preliminarily carried out: 1) the stage of alkylation of the nitrogen atom of dihydroquinoline with methyl iodide, 2) interaction with sulfur gives 4,4,5-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4- c]quinoline-1-thione.

To a solution of 0.01 mol of 2,2,4-trimethyl-1,2-dihydroquinoline in 100 ml of acetone was added 0.015 mol of methyl iodide and boiled for 10 hours, cooled, filtered off 1,2,2,4-tetramethyl-1, 2-dihydroquinoline (preliminary stage 1), 0.01 mol of which, after drying, was boiled with 0.05 mol of molecular sulfur in 30 ml of N,N-dimethylformamide for 12 hours, cooled, the reaction mixture was poured into water, the precipitate was filtered off, dried, the obtained 4,4,5-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione was recrystallized from a mixture of toluene-isopropyl alcohol (3:1) (preliminary stage 2). To a solution of 0.01 mol of 4,4,5-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione in 100 ml of acetone was added 0.015 mol of methyl iodide and boiled for 5 hours, cooled, filtered off the methylthiodithiolium salt, 0.01 mol of which, after drying, boiled with 0.011 mol of N-((4-aminophenyl)sulfonyl)acetamide and 0.02 mol of triethylamine in 100 ml of dry butanol for 3 hours, cooled, filtered off, the solvent was distilled off under reduced pressure, the precipitate was filtered off, recrystallized from dioxane. Yield 65%.

Yellow powder, melting point 247-248°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.40 (br. s, 6H, (CH ₃ ) ₂ ), 1.93 (s, 3H, CH ₃ CO), 2.20 (s, 3H, CH ₃ ) , 2.78 (s, 3H, NCH ₃ ), 6.76 (d, J=8.36 Hz, 1H, H-6(7)); 7.08 (d, J=8.36 Hz, 1H, H-7(6)), 7.24 (d, J=8.60 Hz, 2H, H-Ar), 7.94 (d, J=8.60 Hz, 2H, H-Ar) , 8.39 (s, 1H, H-9), 12.05 (br. s, 1H, CONH); HPLC-HRMS (ESI) calculated for C ₂₂ H ₂₃ N ₃ O ₃ S ₃ +H ⁺ , 474.0976; found, 474.0980.

EXAMPLE 15

Synthesis of (1-((4-ethoxyphenyl)imino)-4,4,8-trimethyl-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)(furan-2- yl)methanone (Io) was carried out via route C.

To a solution of 0.01 mol of 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione in 100 ml of acetone was added 0.015 mol of methyl iodide and boiled for 3 hours, cooled, filtered off the methylthiodithiolium salt, 0.01 mol of which, after drying, boiled with 0.011 mol of 4-ethoxyaniline and 0.02 mol of triethylamine in 100 ml of dry butanol for 3 hours, cooled, filtered off, the solvent was distilled off at reduced pressure, the precipitate was filtered off, recrystallized from isopropyl alcohol. A mixture of 0.01 mol of the resulting imine 4,4,8-trimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 0.012 mol of furan-2-carbonyl chloride was boiled in 100 ml of a mixture of anhydrous toluene and pyridine (10:1) for 5 hours, the pyridinium salt was filtered off, the filtrate was evaporated, the precipitate that formed was filtered off and recrystallized from toluene. Yield 82%.

Yellow powder, melting point 203-204°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.32 (t, 3H, J=6.95 Hz, CH ₃ CH ₂ O), 1.69 (br. s, 6H, (CH ₃ ) ₂ ), 2.23 ( s, 3H, CH ₃ ), 4.02 (q, 2H, J=6.95 Hz, CH ₂ O), 6.65 (d, J=8.15 Hz, 1H, H-6(7)); 6.87 (d, J=8.15 Hz, 1H, H-7(6)), 6.96 (t, J=4.36 Hz, 1H, H-Het), 7.00 (d, J=8.96 Hz, 2H, H-Ar) , 7.06 (d, J=8.96 Hz, 2H, H-Ar), 7.16 (d, J=4.36 Hz, 1H, H-Het), 7.74 (d, J=4.36 Hz, 1H, H-Het), 8.45 (s, 1H, H-9); HPLC-HRMS (ESI) calculated for C ₂₆ H ₂₄ N ₂ O ₃ S ₂ + H ⁺ , 477.1302; found, 477.1300.

EXAMPLE 16

Synthesis of N-(3,4-dichlorophenyl)-8-ethoxy-1-((4-isopropylphenyl)imino)-4,4-dimethyl-1H-[1,2]dithiolo[3,4-c]quinoline-5 (4H)-carboxamide (Ip)

0.015 mol of iodide methyl and boiled for 5 hours, cooled, filtered off the methylthiodithiolium salt, 0.01 mol of which after drying was boiled with 0.011 mol of 4-isopropylaniline and 0.02 mol of triethylamine in 100 ml of dry butanol for 3 hours, cooled, filtered off, the solvent was distilled off under reduced pressure, the precipitate was filtered off, recrystallized from isopropyl alcohol. A mixture of 0.01 mol of the obtained imine 8-ethoxy-4,4-dimethyl-4,5-dihydro-1H-[1,2]dithiolo[3,4-c]quinoline-1-thione and 0.011 mol of 3,4- dichlorophenylisocyanate was boiled in 100 ml of anhydrous dioxane for 6 hours, the precipitate was filtered off, washed with water, dried, and recrystallized from toluene. Yield 80%.

Yellow powder, melting point 189-190°C; ¹ H NMR (DMSO-d ₆ , 500 Hz) δ ppm: 1.22 (d, 6H, J=6.79 Hz, 2 CH ₃ ), 1.29 (t, 3H, J=6.85 Hz, CH ₃ CH ₂ O), 1.65 (br. s, 6H, (CH ₃ ) ₂ ), 2.92 (m, 1H, CH), 3.97 (q, 2H, J=6.85 Hz, CH ₂ O), 6.85 (s, 2H, CH ₃ , H- 6+H-7); 7.01 (d, J=8.10 Hz, 2H, H-Ar), 7.34 (d, J=8.10 Hz, 2H, H-Ar), 7.42 (d, J=8.76 Hz, 1H, H-Ar), 7.55 ( e, J=8.76 Hz, 1Н, Н-Ar), 7.89 (s, 1Н, Н-Ar), 8.29 (s, 1Н, Н-9), 10.14 (br. s, 1H, CONH); HPLC-HRMS (ESI) calculated for C ₃₀ H ₂₉ Cl ₂ N ₃ O ₂ S ₂ + H ⁺ , 598.1152; found, 598.1150.

Compounds with other variants of the radicals given in the general formula I have also been synthesized, which have demonstrated similar activity in inhibiting the replication of betacoronaviruses, including SARS-Cov-2.

EXAMPLE 17

Study of the activity of antiviral chemically synthesized compounds against SARS coronavirus - CoV- 2 in vitro

Chemically synthesized compounds were dissolved in dimethyl sulfoxide (DMSO) to a concentration of 10 mg/ml (stock solution) and stored at -20°C.

In this study, a SARS-CoV-2 coronavirus strain homologous to a viral isolate isolated at the beginning of the pandemic in Wuhan (PRC) was used.

The study of antiviral activity was carried out using a transplanted Vero cell line (epithelial from the kidney of the African green monkey). Cells were grown in Dulbecco's Modified Medium (DMEM) with 10% fetal calf serum, 1% mixture of antibiotics and antimycotics, and 1% glutamine (Gibco, Life Technologies, UK).

Vero cells were seeded in 96-well plates (approximately 2×10 ⁴ cells per well) and incubated for 24 h at 37° C. until a confluent monolayer was formed. The medium was then removed from the wells and replaced with 200 µl of fresh medium containing 0-100 µM compound and incubated for 2 hours. The medium was then removed from the wells and replaced with fresh medium containing virus with a MOI of 0.1. After a 1 hour incubation, the medium was replaced with fresh compound containing medium and incubated for 72-96 hours at 37° C. until a cytopathic effect (CPE) appeared in virus control (no inhibitor) wells.

In all experiments, the virus-infected cells were added as a negative control at a concentration corresponding to the dilution of the initial solution of the compound - DMSO.

The number of viable cells (protected from CPE virus compound) was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT). The test is based on the conversion of pale yellow 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (Sigma-Aldrich, USA) into purple formazan by the enzyme succinate dehydrogenase.

To do this, add an MTT solution (50 µl, 5 mg/ml) to the cell supernatant.

The tablet was incubated for 90 min at 37°C.

The supernatant was completely removed, and the cells were fixed with 4% formaldehyde for 30 min.

MTT crystals were dissolved in 1 ml of 96% ethanol for 10 min.

Optical density (extinction) values were measured on a multifunctional xMark spectrophotometer (Bio-Rad, USA) in 96-well plates at a wavelength of 450 nm. Wells not infected with the virus were used as a control for an intact cell monolayer.

The effectiveness of the inhibition of viral reproduction by the studied compound was determined by the formula:

where OP - optical density

For each dilution of the compound, 3 wells of the plate were used, from which the average value was determined.

Table 1 shows the results of determining the activity of the synthesized compounds against SARS-CoV-2 coronavirus, Remdesevir was tested as a reverse compound.

It can be seen from the above table that at least two compounds, namely 2-(naphthalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[ 3,4-c]quinolin-5(4H)-yl)ethanone Id (EC ₅₀ = 0.51 μM, SI = 411.5) and 2-(naphthalen-2-yloxy)-1-(4,4,7-trimethyl- 1-thioxo-1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)ethanone 1c (EC ₅₀ = 1.16 μM, SI = 86.21), are significantly superior to Remdesavir both in terms of suppression efficiency replication of the SARS-CoV-2 coronavirus, and by the selectivity index, i.e. these compounds have significantly lower cytotoxicity than Remdesavir, with a higher efficiency of suppression of coronavirus replication in cell culture. All these compounds of general formula I remain stable for a long time (1 year) in air at room temperature. Stability analysis was carried out on the basis of data from chromatographic and spectral analysis methods.

Figure 1 shows a graph of the effectiveness of inhibition of replication of the coronavirus SARS-CoV-2 in cell culture Vero E6 and cytotoxicity for the compound 2-(naphtalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo -1H-[1,2]dithiolo[3,4-c]quinolin-5(4H)-yl)ethanone (Id) and reference drug Remdesivir. It can be seen from the graph that the compound 2-(naphthalen-2-yloxy)-1-(4,4,7,8-tetramethyl-1-thioxo-1H-[1,2]dithiolo[3,4-c]quinoline- 5(4H)-yl)ethanone (Id), is significantly superior to Remdesavir both in terms of the effectiveness of suppression of SARS-CoV-2 coronavirus replication and in terms of selectivity index, the cytotoxicity of Id is significantly lower than that of Remdesavir, with a higher efficiency of suppression of coronavirus replication in cell culture.

EXAMPLE 18.

To obtain a drug in the form of tablets, 1600 mg of starch, 1600 mg of crushed lactose, 400 mg of talc and 1000 mg of compound Id are mixed, compressed into a bar. The resulting bar is crushed into granules and sifted through sieves, collecting granules with a size of 14-16 mesh. The resulting granules are tabletted into a suitable tablet form weighing 560 mg.

EXAMPLE 19.

To obtain a drug in the form of capsules, compound Id is thoroughly mixed with lactose powder in a ratio of 2:1. The resulting powder mixture is packaged in 250 mg gelatin capsules of a suitable size.

EXAMPLE 20.

Obtaining a medicinal product in the form of injectable compositions for intramuscular, intraperitoneal or subcutaneous administration.

Mix 500 mg of compound Id with 300 mg of chlorobutanol, 2 ml of propylene glycol and 100 ml of injection. The resulting solution is filtered and placed in 1 ml ampoules and sealed.

Claims (7)

1. The use of a tricyclic sulfur-containing derivative of 1,2-dihydroquinoline of general formula I to suppress the replication of beta-coronavirus:

where R1 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R2 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R3 is a substituent selected from methyl, ethyl, methoxy, ethoxy; R4 is a substituent selected from methyl-, benzyl-, 3-phenylacryloyl-, 3-(4-methoxyphenyl)acryloyl-, 3-phenylpropanoyl-, phenylethyl, 2-(4-fluorophenoxy)acetyl-, 4-chlorophenoxyacetyl-, 2-(2,4-dichlorophenoxy)acetyl-, 4-nitrobenzoyl-, 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl-) groups; 3,4-dichlorophenylcarbamoyl-, furan-2-carbonyl-, ethylcarbamoyl-, 2-naphthylyloxyacetyl-, 2-(1,3-dioxoisoindolin-2-yl)acetyl-, 2-(1,3-dioxohexahydro-1H-isoindole -2(3H)-yl)acetyl-, 2-morpholinoacetyl-, piperidin-1-ylacetyl-, 2-(4-carbamoylpiperidin-1-yl)acetyl-, 2-(2,5-dioxopyrrolidin-1-yl) acetyl-, 2-phenoxyacetyl-, 2-1,3-dioxo-1Н-benzo[de]isoquinolin-2(3H)-yl)acetyl-, 2-benzo[d]oxazol-2-ylthio)acetyl-, 2 -(benzo[d]thiazol-2-yloxy)acetyl-, benzoxazol-2-ylmercaptoacetyl-, furanoyl-, 2-naflinyloxymethylcarbonyl, 3,4-dichlorophenylcarbamoyl; X is a substituent selected from sulfur, unsubstituted or substituted at the positions of 2,3,4,5,6-arylamine selected from the group consisting of 4-(4-chlorobenzyloxy)phenylimino-, 4-isopropylphenylimino-, 4-(6 -methylbenzo[d]thiazol-2-yl)phenylimino-, 4-(N-benzoylsulfamoyl)phenylimino-, 4-ethoxyphenylimino-, 4-ethoxycarbonylphenylimino-, phenylimino-, 4-chlorophenylimino-, 3-chlorophenylimino-, 4-sulfamoylphenylimino -, 4-(N-acetylsulfamoyl)phenylimino-, 4-(N-(thiazol-2-yl)sulfamoyl)phenylimino-, 2-nicotinoylhydrazono-, 4-((4-chlorobenzyl)oxy)phenylimino-, 2-methoxyphenylimino -, 4-(N-(pyrimidin-2-yl)sulfamoyl)phenylimino-, 4-(N-(4,6-dimethylpyrimidin-2-yl)sulfamoyl)phenylimino-. 2. Use according to claim 1, characterized in that the compound of general formula I is used to suppress the replication of SARS-CoV-2. 3. Use according to claim 1, characterized in that the compound of general formula I is used for the treatment of diseases caused by betacoronaviruses. 4. Use according to claim 1, characterized in that the compound of general formula I is used in the complex therapy of diseases caused by betacoronaviruses.

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