Lviv Polytechnic National University
Lviv Polytechnic National University
Lviv Polytechnic National University
Lviv Polytechnic National University
Institute of animal biology of NAAS

The actual problem in medicine is a phenomenon of antibiotic resistance, therefore, it is expedient to develop and introduce new antibiotics or to improve the transport of already known antibiotics into the cell. PEGylating is the one of the most successful ways of improving of the delivery of therapeutic molecules to the cell, such as proteins, oligonucleotides and other biomolecules. Thus, it is promising to introduce PEG fragments into the structure of antibiotics with the preservation of their biological activity.

The object of research is enrofloxacin, an antibiotic of the third generation of fluoroquinolones, that are derivatives of 4-quinolone and contain a piperazine ring and a fluorine atom. The presence of fluorine atom in their structure substantially expands the spectrum of their antibacterial activity. Enrofloxacin is successfully used in veterinary medicine for the treatment of many bacterial diseases, but due to the rapid increase in the resistance of microorganisms to the action of antibiotic, the development of new antibiotics based on enrofloxacin is relevant. The presence of a reactive carboxyl group in the molecule of the antibiotic makes it possible to modify its structure for obtaining new compounds.

During the development of the method for the modified enrofloxacin production, the following reactions were considered: straight esterification of carboxyl groups, esterification using Steglich reaction and obtaining esters through the intermediate formation of chloranhydride. The yield of PEGylated product in straight esterification and using Steglich reaction did not exceed 20-25%, therefore the synthesis of PEGylated enrofloxacin is proposed through the intermediate formation of its chloranhydride. During the reaction the formation of PEGylated enrofloxacin was monitored by IR spectroscopy. The purity of the products according to the data of high-performance liquid chromatography was 98%.

It is important for such a modification that the obtained PEGylated enrofloxacin hadn`t, at least, less bactericidal properties than the original antibiotic. Antibacterial activity of PEGylated compounds was investigated by serial dilutions using Pseudomonas aeruginosa culture. It is shown that antibacterial activity of the enrofloxacin compounds covalently coupled to polyethylene glycol are higher in comparison with the initial antibiotic.

1. Никитин И. Г., Байкова И. Е., Гогова Л. М. Пегилированные лекарственные препараты:
современное состояние проблемы и перспективы // Лечебное дело. – 2005. – № 4. – С. 18–24. 2. Paola
Milla, Franco Dosio and Luigi Cattel. PEGylation of proteins and liposomes, a powerful and flexible strategy to improve the drug delivery // Current Drug Metabolism. – 2012. – Vol.13, 1. – Р. 105–119(15). 3. Gianfranco
Pasut, Mauro Sergi, Francesco M. Veronese. Anti-cancer PEG-enzymes: 30 years old, but still a current
approach // Advanced Drug Delivery Reviews, 60. – 2008. – Р. 69–78. 4. Omolo, Rahul S. Kalhapure Mahantesh Jadhav, Sanjeev Rambharose, Chunderika Mocktar. Valence M. K. Ndesendo, Thirumala Govender. Pegylated oleic acid: A promising amphiphilic polymer for nanoantibiotic Delivery // European Journal of Pharmaceutics and Biopharmaceutics. – 2017. – 112. – Р. 96–108. 5. Gregory Marslin, Ann Mary Revina, Vinoth Kumar Megraj Khandelwal, Krishnamoorthy Balakumar, Caroline J. Sheeba, Gregory Franklin. PEGylated ofloxacin nanoparticles render strong antibacterial activityagainst many clinically important human pathogens // Colloids and Surfaces B: Biointerfaces. – 2015. – Vol. 132. – Р. 62–70. 6. N. Elmarzugi, T. Adali, A. Bentaleb, E. Keleb, A. Mohamed, A. Hamza. Spectro-scopic characterization of PEG–DNA biocomplexes by FTIR // J. Appl. Pharm. Sci. – 2014. – 4 (08). – Р. 6–10. 7. E. Froehlich, J.S. Mandeville, C.M. Weinert, L. Kreplak, H.A. Tajmir-Riahi // Bundling and aggregation of DNA by cationic dendrimers // Biomacromolecules. – 2011. – 12. – Р. 511–517. 8. Gabor Pinter, Pal Horvath, Sandor Bujdoso, Ferenc Sztaricskai, Sandor Keki, Miklos Zsuga, Szilvia Kardos, Ferenc Rozgonyi and Pa´l Herczegh. Synthesis and antimicrobial activity of ciprofloxacin and norfloxacin permanently bonded to polyethylene glycol by a thiourea linker // The Journal of Antibiotics. – 2009. – 62. – Р. 113–116. 9. Yu-SenWang, Stephen Youngster, James Bausch, Rumin Zhang, Charles McNemar, and Daniel F. Wyss Identification of the Major Positional Isomer of Pegylated Interferon Alpha // Biochemistry. – 2000. – 39. – Р. 10634–10640. 10. Christian F. Jehn, Philipp Hemmati, Silvia Lehenbauer-Dehm, Sherko Kümmel ,Bernd Flath, Peter Schmid. Biweekly Pegylated Liposomal Doxorubicin (Caelyx) in Heavily Pretreated Metastatic Breast Cancer: A Phase 2 Study // Clinical Breast Cancer. – Vol. 16, No. 6. – Р. 514. 11. Tessa Trouchon, Sébastien Lefebvre. A Review of Enrofloxacin for Veterinary Use, Scientific Research Publishing Inc. - USC 1233 INRA. - Vetagro Sup, Veterinary School of Lyon. – Marcy l’Etoile. – France, 2016. – Р. 20. 12. Ковалев В. Ф., Волков И. Б., Виолин Б. В. и др. Антибиотики, сульфаниламиды и нитрофураны в ветеринарии. –М.: Агропромиздат, 1988. – 223 c. 13. B. Chakrabarty, A.K. Ghoshal, M.K. Purkait. Effect of molecular weight of PEG on membrane morphology and transport properties // Journal of Membrane Science. – 2008. – Vol. 309. –
Р. 209–221.