IBCh positions

2021–to date

Leading research fellow

Laboratory of biosynthesis of physiologically active compounds

Titles

Doctor of Philosophy (Chemistry)

Scientific results

In last ten years Konstantinova Irina  directly manages the complex of works on development of chemical-enzymatic method of obtaining new modified nucleosides. Defines the possible directions of work on modification of structures of known nucleosides with the aim of getting promising compounds with unusual set of properties. Those compounds are of great interest for oncology and virology. Professionally at a high level owns by NMR spectroscopy in determining the structure of new synthesized compounds. She has for the first time proved formation of regioizomer at synthesis of nucleosides of benzimidazole [1,2,4,9]. A critical attitude to the results of experiments allows her to notice atypical behavior of enzymes in synthesis of new compounds [3] that led to the discovery of some evidences for the hypothesis about the key influence of the serine-90 on the formation of the substrate –enzyme complex of the active center of purine nucleoside phosphorylase.

Under the leadership of I.D. Konstantinova effective domestic biocatalytic technologies of receiving substances of pharmaceuticals on the basis of the modified nucleosides have been developed: cladribin, fludara, nelarabin, vidarabin, ribavirin. Irina D. Konstantinova  is the author of 3 patents of the Russian Federation for technologies of receiving substances of these medicines.

With group of coauthors I.D. Konstantinova  performed the analysis of metabolic transformations of carbohydrates and nucleosides in the active centers of enzymes. Those are the most important enzymatic transformations of the specified compounds in terms of their possible use in biotechnology.  Among enzymes of nucleic acid metabolism the attention has been focused on ribokinase (RK), phosphopentomutase (FPM) and nucleosidephosphorylases (namely, uridinephosphorylase (UF), a thymidinephosphorylase (TF), and purinenucleosidephosphorylase (PNF). It was shown that kinases, mutases, and nucleosidephosphorylases  E. coli are of the greatest interest.

The detailed analysis of operating conditions of all enzymes and their substrate specificity was carried out. This allowed to formulate the original strategy of receiving biologically important nucleosides which consists in use of RK, FPM, and nucleosidephosphorylase "in one pot". The initial pentose consistently turns under the action of RK in 5-phosphate, then in 1-phosphate under the action of FPN, and finally, in nucleosides by the condensation of 1-phosphates with heterocyclic bases catalyzed by nucleosidephosphorylases. As the pentoses, in this cascade transformation can be used D–ribose, 2-deoxyribose, 3-deoxyribose, arabinose, xylose, 2-deoxy-2-fluoro-ribose, and 2-deoxy-2-fluoro-arabinose.

It was found that synthetic analogues of the natural purines and pyrimidines can be used  as heterocyclic bases [5,6,7].

The developed methodology of cascade enzymatic synthesis in one pot has no analogues in the world scientific and patent literature. Konstantinova is the winner of the RAS-Belarus NAS Award 2015 for a series of joint works with Institute of Bioorganic chemistry of Belarus National Academy of Sciences on the topic: "A multienzyme cascade in the conversion of carbohydrates nucleosides: a new strategy for the synthesis of biologically important nucleosides".

Under the direct supervision of Konstantinova metabolically stable analogues of nucleoside antibiotic cordycepin were received. Those compounds are of great interest for the treatment of human African trypanosomiasis [African Human trypanosomiasis (HAT)] [10].

The research results became the basis for the development of scalable technology to produce a number of biologically important compounds (clofarabine, fludarabine, cladribine, ribavirin, nelarabin, etc.). Those compounds were synthsized in preparative amounts (tens and hundreds of grams) to conduct medical and biological tests for registration them in the Russian Federation as medical products. In addition, a detailed analysis of both own results and literary data on the structure of catalytic centers nucleosidephosphorylases allowed to clarify the mechanism of the "recognition" of the heterocyclic bases by mentioned enzymes. This work is under development, however already today we have proposed new structures of heterocyclic bases and nucleosides, electronic properties of which will be expected of substrate or inhibitory activity.

I. D. Konstantinova is author of four patents for new of modified nucleosides. Personally corresponds with the FIPS examiners, skillfully resolves conflict situations in patenting.

Patents:

- Konstantinova I. D., Eletskaia B. Z, Dorofeeva, E. V., Fateev V. I., Berzina, M. J., Lutonina O. I., Miroshnikov A. I. "Method of production of 9-beta-D-arabinofuranosyl-6-(Nα-L-serylamido)-2-chloropurine". RF patent № 2563257, 21.08.2015 Patent priority 07.11.2014. Application RF № 2014144731, 07.11.2014 G.

- Konstantinova I. D., Fateev I. V., Miroshnikov, A. I., "A method of purine nucleosides of β-D-arabinofuranose". Patent application RF № 2015147715, 06.11.2015 Decision on patent grant 30.03.2017 Patent RF № 2624023, 30.06.2017, priority 06.11.2015.

- Konstantinova I. D., Kharitonova, M. I., Miroshnikov A. I., Kotovskaya S. K., Charushin V. N., Galegov G. A., Andronova V. L. "2-Amino-5,6-difluoro-1-(beta-D-ribofuranosyl)-benzimidazole, the method of obtaining and antiviral activity against herpes simplex virus Type 1" Patent Application No. 2016132163/04 (049868), 04.08.2016, Decision on granting of a patent 1 0.07.2017.

- Konstantinova I. D, Chudinov  M. V. , Prutkov A. N., Matveev A. V., Grebenkina L. E. Dorofeeva E. V. "Amide 5-(tetrahydrofuran-2-yl)-1,2,4-triazole-3-carboxylic acid, having antiviral activity, and method for its production". Patent application RF № 2016138180, 26.09.2016 Decision on patent grant 25.05.2017.

Currently, Konstantinova I. D. is preparing her doctoral thesis. In the Laboratory of Biotechnology of Institute of Bioorganic Chemistry she determines the further development trends in the synthesis of modified oligonucleotides, directly administers the experimental work of seven members of the Laboratory of biotechnology, three graduate students, and five students of Moscow colleges.

1. Fateev, I.V.; Antonov, K.V.; Konstantinova, I.D.; Muravyova, T.I.; Seela, F.; Esipov, R.S.; Miroshnikov, A.I.; Mikhailopulo, I.A. The chemoenzymatic synthesis of clofarabine and related 2′-deoxyfluoroarabinosyl nucleosides: the electronic and stereochemical factors determining substrate recognition by E. coli nucleoside phosphorylases // Beilstein J. Org. Chem. 2014, 10, pp. 1657-1669. DOI: 10.3762/bjoc.10.173

2.I. V. Fateev, M. I. Kharitonova, K. V. Antonov, I. D. Konstantinova, V. N. Stepanenko, R. S. Esipov, F. Seela, K. W. Temburnikar, K. L. Seley-Radtke, V. A. Stepchenko, Y. A. Sokolov, A. I. Miroshnikov, I. A. Mikhailopulo. Recognition of the Challenging Heterocyclic Bases by E. coli Purine Nucleoside Phosphorylase vs its Ser90Ala Mutant. // Chem. Eur. J., 2015, V. 21 (38). P. 13401-13419. DOI: 10.1002/chem.201501334

3. M. I. Kharitonova, I. V. Fateev, A. L. Kaushin, I. D. Konstantinova, S. K. Kotovskaya, V. L. Leont’eva, G. A. Galegov, V. N. Charushin, A. I. Miroshnikov. Chemo-Enzymatic Syntheses and Antiviral Evaluation of 5-substituted 4,6-difluorobenzimidazoles Ribo- and 2’-Deoxyribo-Nucleosides // Synthesis, 2016; 48(03): 394-406, DOI: 10.1055/s-0035-1560911

4. B. Z. Eletskaya, I. D. Konstantinova, A. S. Paramonov, R. S. Esipov, D. A. Gruzdev, A. Yu. Vigorov, G. L. Levit,  A. I. Miroshnikov, V. P. Krasnov and V. N. Charushin. Chemoenzymatic arabinosylation of 2-aminopurines bearing the chiral fragment of 7,8-difluoro-3-methyl-3,4-dihydro-2H-[1,4]benzoxazines //  Mendeleev Commun., 2016, 26, p. 6−8.

5. M. V. Chudinov, A. N. Prutkov, A. V. Matveev, L. E. Grebenkina, I. D.Konstantinova, Y. V. Berezovskaya. An alternative route to the arylvinyltriazole nucleosides.// Bioorg. Med. Chem. Lett. 2016. V. 26, р. 3223-3225.   DOI: 10.1016/j.bmcl.2016.05

6. Chudinov, M.V., Matveev, A.V., Prutkov, A.N.,  Konstantinova,I.D., Fateev, I.V., Prasolov, V.S., Smirnova, O.A., Galegov, G.A., Deryabin, P.G. Novel 5-alkyl(aryl)-substituted ribavirine analogues: Synthesis and antiviral evaluation. // Mendeleev Communications V. 26 (3), 2016, pp. 214-21

7. Konstantinova, I.D., Fateev, I.V., Miroshnikov, A.I.  The arsenolysis reaction in the biotechnological method of synthesis of modified purine β-D-arabinonucleosides. // Russian Journal of Bioorganic Chemistry, 2016, 42 (4), 372-380

8. M. I. Kharitonova, I. V. Fateev, K.V.Antonov, A. L. Kaushin, I. D. Konstantinova, S. K. Kotovskaya, V. L. Andronova, G. A. Galegov, V. N. Charushin, A. I. Miroshnikov. Chemo-Enzymatic Syntheses and Antiviral Evaluation of 5-substituted 4,6-difluorobenzimidazoles 2’-fluoro-arabinosides // Synthesis 2017, V. 49, pp. 1043-1052. DOI 10.1055/s-0036-1588625

9. M. I. Kharitonova, A. O. Denisova, V. L. Аndronova, A.L. Kayushin, I.D. Konstantinova, S.K. Kotovskaya, G.A. Galegov, V.N. Charushin, A.I. Miroshnikov. New modified 2-aminobenzimidazole nucleosides: synthesis and evaluation of their activity against herpes simplex virus type 1// Bioorg. Med. Chem. Lett., 2017, V. 27, p. 2484-2487. DOI 10.1016/j.bmcl.2017.03.100

10. Denisova A.O., Tokunova Y.A., Fateev I.V., Breslav A.A., Leonov V.N., Dorofeeva E.V., Lutonina O.I., Muzyka I.S., Esipov R.S., Kayushin A.L., Konstantinova I.D., Miroshnikov A.I., Stepchenko V.A., Mikhailopulo I.A. The Chemoenzymatic Synthesis of 2-Chloro- and 2-Fluorocordycepins // Synthesis, 2017, V. 49, DOI: 10.1055/s-0036-1590804