Design of new pharmacophore molecules, which contain vital phosphorus-containing heterocycles and their derivatives, is one of the urgent tasks of bioorganic chemistry. In the present paper, we report the original data on the synthesis and bactericidal properties of fluorine-containing dioxaphosphorinane oxides, representatives of the biologically important class of organic cyclic phosphates (nucleotides, phosphorylated carbohydrates, phospholipids), which play an important role in biochemical processes. In recent years, these compounds have been widely studied as promising precursors of drugs, monomers for the production of biomedical materials, ligands for metal complexes, as well as building blocks for organic synthesis. Among them are substances with antibacterial, antiviral, fungicidal and insecticidal activity. Special attention is paid to the directed synthesis of phosphorus-containing heterocycles with pharmacophoric fluoroalkyl fragments. In this study, we report on the synthesis of some representatives of dioxaphosphorinanе oxides with exocyclic fluoroalkyl substituents. The target dioxaphosphorinane oxides were obtained by us in two stages based on the available reagents (phosphoryltrichloride, fluorinated alcohols and 1,3-butanediol) under mild experimental conditions. At the first stage, the synthesis of trifluoroethyl, tetrafluoropropyl and octafluoropentyl dichlorophosphates from phosphoryltrichloride and fluorinated alcohols is carried out in the presence of catalytic amounts of lithium chloride. In the second stage, cyclization of polyfluoroalkyl dichlorophosphates with 1,3-butanediol was used to construct six-membered phosphorus-containing heterocycles. The reaction readily proceeds at a temperature of –10 ¸ 25˚C for 5 h in the presence of pyridine in diethyl ether and leads to the formation of 4-methyl-2-(polyfluoroalkoxy)-1,3,2-dioxaphosphorinanе 2-oxides with a preparative yield of up to 71%. 4-Methyl-2-(2,2,2-trifluoroethoxy)-1,3,2-dioxaphosphorinanе 2-oxide was obtained by us for the first time in this work according to the developed technique. The structure and composition of the synthesized individual representatives of the class of cyclic organic phosphates - polyfluoroalkylated phosphorinanе oxides has been reliably proven based on NMR spectroscopy data on ¹H, ¹³C, ¹⁹F, ³¹P nuclei, IR spectroscopy and elemental analysis. The use of two-dimensional homo- and heteronuclear NMR spectroscopy (COZY, HSQC, HMBC) made it possible to confirm the existence of synthesized 4-methyl-2-(polyfluoroalkoxy)phosphorinane oxides in the form of two isomers with a cis- and trans- location of a polyfluoroalkoxy substituent at the phosphorus atom and the methyl group in 4 the position of the phosphorus-containing cycle (isomer ratio ~ 1:1). The bactericidal activity of the obtained cyclic phosphates was evaluated by the agar well diffusion method with respect to the strains of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus and Bacillus thuringiensis. Fish peptone agar was used for work, 0.02 mmol of the active substance was added to the well in the form of an aqueous solution (for trifluoroethoxy and tetrafluoropropoxy substituents for dioxaphosphorinane oxides) or an emulsion (for an octafluoropentyloxy group with dioxaphosphorinane oxide). Cultures were incubated at +30°C, and after 24 hours, results were recorded. Test-cultures under similar conditions without adding the studied substances to the wells was used as a control. Studies have shown that a previously unknown dioxaphosphorinane oxide with a trifluoroethoxy group has a bactericidal effect on E. coli and P. aeruginosa strains, while compounds with longer polyfluoroalkoxy substituents (with tetrafluoropropoxy and octafluoropentyloxy groups) do not inhibit the growth of these cultures. At the same time, a 10-fold decrease in the concentration of dioxaphosphorinane oxide with the trifluoroethoxy group (0.002 mmol of the active substance) also has a bactericidal effect on the E. coli strain. It should be noted that, in the P. aeruginosa strain, there is an intensive production of pigment compared with the control in the case of using all the studied phosphorinane oxides, regardless of the degree of fluorination of the polyfluoroalkoxyl substituent. Strain S. aureus showed sensitivity to all three polyfluorine-containing compounds, while there is a bacteriostatic effect. The synthesized compounds do not affect the growth of B. cereus soil bacterial strains (which can cause toxic humans infections) and the entomopathogenic B. thuringiensis bacterium. The data obtained evidence that fluorine-containing dioxaphosphorinane oxides could be successfully used as agents that inhibit the growth of conditionally pathogenic bacteria such as P. aeruginosa and S. aureus, causing opportunistic humans infections.

Mikhailenko Valentina Lvovna, Candidate of Science (Chemistry), Associate Professor, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation, tel.: (3952) 24–18–55, e-mail: mival63@gmail.com 

Vyatchina Olga Fedorovna, Candidate of Science (Biology), Associate Professor, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation, tel.: (3952) 24–18–55, e-mail: olgairk3@rambler.ru 

Nalibayeva Araylym Muratovna, Research Scientist, D.V. Sokolsky Institute of Fuel, Catalysis and Electrochemistry, 142, Kunayev st., Almaty, 050010, Kazakhstan, tel.: (727) 291–65–58, e-mail: aray77@mail.ru 

Pozdeeva Aleksandra Sergeevna, Graduate Student, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation, tel.: (3952) 24–18–55 

Bishimbayeva Gaukhar Kozykeevna, Doctor of Science, Professor, Principal Research Scientist, D.V. Sokolsky Institute of Fuel, Catalysis and Electrochemistry, 142, Kunayev st., Almaty, 050010, Kazakhstan, tel.: (727) 291-58-08, e-mail: gaukhar.bishimbayeva@gmail.com 

Verkhoturova Svetlana Ilyasovna, Candidate of Science (Chemistry), Senior Research Scientist, A. E. Favorsky Irkutsk Institute of Chemistry SB RAS, 1, Favorsky st., Irkutsk, 664033, Russian Federation, tel.: (3952) 42–59–31, e-mail: verkhoturova@irioch.irk.ru

Mikhailenko V.L., Vyatchina O.F., Nalibayeva A.M., Pozdeeva A.S., Bishimbayeva G.K., Verkhoturova S.I. Estimation of Bactericidal Activity of Fluorine-Containing Dioxaphosphorinane Oxides. The Bulletin of Irkutsk State University. Series Biology. Ecology, 2019, vol. 27, pp. 30-40. https://doi.org/10.26516/2073-3372.2019.27.30 (in Russian)

bactericidal activity, fluorine-containing dioxaphosphorinane oxides, 4-methyl-2-(2,2,2-trifluoroethoxy)-1,3,2-dioxaphosphorinane-2-oxide, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus, Bacillus thuringiensis

Netrusov A.I. Praktikum po mikrobiologii [Workshop on microbiology]. Moscow, Akademia Publ., 2005, 608 p. (in Russian)

Gusarova N.K., Verkhoturova S.I., Arbuzova S.N., Kazantseva T.I., Nalibayeva A.M., Bishimbayeva G.K. Sintez tsianoetilirovannykh ftoralkilfosfatov [Synthesis of cyanoethylated fluoroalkylphosphates]. Butlerov Communications, 2016, vol. 47, no. 8, pp. 29-34. (in Russian)

Gusarova N.K., Verkhoturova S.I., Arbuzova S.N., Kazantseva T.I., Albanov А.I., Nalibayeva A.M., Bishimbayeva G.K. Sintez poliftoralkilirovannykh 1,3,2-dioksafosfolan- i 1,3,2-dioksafosforinanoksidov [Synthesis of Polyfluoralkylated 1,3,2-Dioxaphospholane and 1,3,2-Dioxaphosphorinane Oxides]. Rus. J. Org. Chem., 2017, vol. 53, no. 11, pp. 1623-1629. https://doi.org/10.1134/S107042801711001X. (in Russian)

Carroll S.S., Koeplinger K., Vavrek M., Zhang N.R., Handt L., MacCoss M., Olsen D.B., Reddy K.R., Sun Z., van Poelje P.D., Fujitaki J.M., Boyer S.H., Linemeyer D.L., Hecker S.J., Erion M.D. Antiviral Efficacy upon Administration of a HepDirect Prodrug of 2`-C-Methylcytidine to Hepatitis C Virus-Infected Chimpanzees. Antimicrobial agents and chemotherapy, 2011, vol. 55, no. 8, pp. 3854-3860. https://doi.org/10.1128/AAC.01152-10

Baszczyňski O., Janeba Z. Medicinal Chemistry of Fluorinated Cyclic and Acyclic Nucleoside Phosphonates. Med. Res. Rev., 2013, vol. 33, no. 6, pp. 1304-1344. https://doi.org/10.1002/med.21296

Liao S., Fan S.-Y., Liu Q., Li C.-K., Chen J., Li J.-L., Zhang Z.-W., Zhang Z.-Q., Zhong B.-H., Xie J.-W. In vitro evaluation of 9-(2-phosphonylmethoxyethyl)adenine ester analogues, a series of anti-HBV structures with improved plasma stability and liver release. Arch. Pharm. Res., 2014, vol. 37, pp. 1416-1425. https://doi.org/10.1007/s12272-013-0300-6

Duda A., Kubisa P., Lapienis G., Slomkowski S. Milestones in development of a ring-opening polymerization of the heterocyclic monomers - view from a personal perspective. Polimery, 2014, vol. 59, no. 1, pp. 9-23. http://dx.doi.org/10.14314/polimery.2014.009

Dadapeer E., Naidu K.R.M., Ramesh M., Raju C.N., Devamma M.N. Synthesis and biological activity of alkyl-2-[5-(hydroxy methyl)-5-nitro-2-oxo-1,3,2λ⁵-dioxaphosphinan-2-yl]amino acid esters. J. Chem. Pharm. Res., 2010, vol. 2, no. 3, pp. 109-116. 

Shi D.Q., Liu Y., Feras A.D., Tan X.S., Chen J.X. Synthesis and Biological Activity of Cis 2-(6-Chloropyridine-3-yl)methylamino-4-substitutedphenyl-5,5-dimethyl-1,3,2-dioxaphosphinane 2-Oxides. Phosphorus, Sulfur, and Silicon, 2005, vol. 180, no. 8, pp. 1937-1946. http://dx.doi.org/10.1080/104265090902697

Pradere U., Garnier-Amblard E.C., Coats S.J., Amblard F., Schinazi R.F. Synthesis of Nucleoside Phosphate and Phosphonate Prodrugs. Chem. Rev., 2014, vol. 114, no. 18, pp. 9154-9218. https://pubs.acs.org/doi/10.1021/cr5002035

Wiemer A.J., Wiemer D.F. Prodrugs of Phosphonates and Phosphates: Crossing the Membrane Barrier. Top. Curr. Chem., 2015, vol. 360, pp. 115-160. https://doi.org/10.1007/128_2014_561