1 IN VITRO ANTIOXIDANT ACTIVITY OF COMPOSITIONS BASED ON BIOSURFACTANTS AND AMINO- CONTAINING HETEROCYCLIC DERIVATIVES OF 1,4-NAPHTOQUINONE Nataliia Polish *1 , Olena Yaremkevych 1 , Nataliia Marintsova 1 , Lesia Zhurakhivska 1 , Nataliia Koretska 2 , Tetyana Pokynbroda 2 , Nataliia Liubas 3 , Vira Lubenets 1 , Elena Karpenko 2 Address(es): 1 Lviv Polytechnic National University, Department of Technology Biologically Compounds, Pharmacy and Biotechnology, 3/4 St. George Square, Lviv, Ukraine, 79013. 2 Department of Physical Chemistry of Fossil Fuels of the Institute of Physical-Organic Chemistry and Coal Chemistry named after L. M. Lytvynenko of the National Academy of Sciences of Ukraine, Str. Naukova, 3a, Lviv, Ukraine, 79060. 3 Institute of Animal Biology, NAAS of Ukraine, V. Stus Str., Lviv, Ukraine, 79034. *Corresponding author: polishn@ukr.net ABSTRACT Keywords: antioxidant activity, amino-containing heterocyclic 1,4-naphthoquinone derivatives, rhamnolipids, trehalosе lipids, GUSAR INTRODUCTION The increase in the processes of free-radical oxidation in the body is observed in many diseases. Their influence in the pathogenesis of various types of cancer, atherosclerosis, hypertension, Alzheimer's disease, Parkinson's disease (Cassidy, et al., 2020; Jiang, et al., 2016), arthritis, neurodegenerative disorders, diabetes has been proven (Li, et al., 2016; Liu, et al., 2017). Antioxidants (AO) are the main group of drugs that can resist oxidative stress. They are able to inhibit the development of free-radical reactions, preventing the formation of peroxides that damage the cellular and subcellular membranes of the body (Karuppagounder, et al., 2021). That is why the need to find new AO that would be effective in treatment of number of diseases is an important task of scientists for today. Heterocyclic amino-containing derivatives of 1,4-naphthoquinone and their complexes with TL and RL biosurfactants were choosen for the study of antioxidant activity. Literary sources suggest that 1,4-naphthoquinone derivatives have redox properties. They affect molecular mechanisms of oxygen exchange in tissues and are able to regulate the flow of electrons in the respiratory chain (Gulcin, et al., 2020). This indicates the possibility of their use as AO. 1,4- Naphtoquinones are poorly soluble in water, which makes their use difficult. Interfering with the solubilization properties, as well as the ability to increase the permeability of cell membranes, biosurfactants increase the bioavailability and activity of biologically active substances. RL and TL belong to low molecular glycolipid biosurfactants (Jahan, et al, 2020). Due to the amphiphilic nature of the molecules, biosurfactants contribute to the solubilization of poorly soluble substances, improve the permeability of cell membranes and enhance the effect of biologically active substances when they are used in combination (Ceresa, et al., 2021; Sotirova et al., 2012) and are low-toxic. RL are products of biosynthesis of Pseudomonas bacteria, where rhamnose is attached to the fatty acid tail by a glycosidic bond (Haba, et al., 2014). TL are biosynthesis products of actinomycetes bacteria, containing trehalose disaccharide linked to mycolic acids (Semeniuk, et al., 2022). The physicochemical properties of biosurfactants contribute to increasing the effectiveness and stabilizing the functional properties of drugs for various purposes (Shekhar et al., 2015). Literature sources suggest that biosurfactants can be used in pharmacy (Naughton, et al., 2019), in particular, as antimicrobials (Zhao et al., 2010; Abdel-Megeed et al., 2011; Sha et al., 2011) , antiviral (Singh et al., 2004; Akbari et al., 2018), antitumor (Pasiar et al., 2016; Yuewen et al., 2017), anti-adhesive drugs (Rodrigues et al., 2006; Bucci et al., 2018; Janek et al., 2018) and as immunological adjuvants (Al-wazni et al., 2016). However, most of the compositions are still at the stage of development and research. Thus, it was determined that the bioavailability and biological (antimicrobial) activity of thiosulfonates significantly increased in the composition with RL (Lubenets et al., 2013). All this information indicates the feasibility of creating compositions of biologically active compounds with biosurfactants (Koretska, et al., 2019). The main goal of creating a composition of synthetic heterocyclic amino derivatives of 1,4-naphthoquinone with biosurfactants is to improve the solubility and reduce the therapeutic dose of the drug. It is known that N-containing heterocyclic derivatives of 1,4-naphthoquinone (Polish et. al., 2020) demonstrate high anticonvulsant activity. However, it is known that they are characterized by low solubility in water, which complicates their use. Due to their ability to regulate the permeability of cell membranes, biosurfactants enhance the effect of biologically active substances and increase the bioavailability of sparingly soluble substances (Banat et al., 2010; Salihu, et al., 2009). This makes it possible to create new drugs with improved functional properties. We have developed compositions of 1,4- naphthoquinone derivatives with RL, which demonstrate high anticonvulsant activity (Polish et. al., 2022). The main goal of the combined use of synthetic naphthoquinone derivatives and biosurfactants is to improve water solubility, bioavailability and reduce the therapeutic dose (inhibitory concentration) of the drug (Sotirova et al., 2012; Koretska et al., 2019). MATERIALS AND METHODS Materials In this study, we used rhamnolipids, i.e., products of microbial synthesis of Pseudomonas sp. PS-17 and TL produced by Rhodococcus erythropolis AU-1. Heterocyclic amino-containing derivatives of 1,4-naphthoquinone (1, 2): 2-[(6-(4- fluorophenyl-5-oxo-2,5dihydro-1,2,4-triazin-3-yl)phenyl)amino]naphthalene - 1,4-dione (1) was obtained by reacting 1,4-naphthoquinone with a derivative of 1,2,4-triazine by the Michael reaction, ethyl-4-((3-chloro-1,4-dioxo-1,4- dihydronaphthalene-2-yl)amino)-1-phenyl-1H-pyrazole-3-carboxylate (2) was Compositions of rhamnolipids (RL) and trehalosе lipids (TL) with amino-containing heterocyclic derivatives of 1,4-naphthoquinone were obtained for the first time. The acute toxicity of amino-containing derivatives of 1,4-naphthoquinone was determined by in silico method using the GUSAR program. We have studied influence of selected compounds and their rhamnolipid and trehalosе lipids compositions on the processes of lipid peroxidation (LPO) and oxidative modification of proteins (OMP) in rat liver tissues upon initiation of free radical oxidation in vitro. It was determined that 1,4-naphthoquinone derivatives have antioxidant activity. RL and TL have a pro-oxidant effect, increase the content of secondary products of lipoperoxidation compared to the control. They have an antioxidant effect in a composition with 1,4-naphthoquinone derivatives. The lead-compound 2-[(6-(4-fluorophenyl-5-oxo-2,5-dihydro-1,2,4-triazin-3- yl)phenyl)amino]naphthalene-1,4-dione (1) and its complex with rhamnolipid show high antioxidant activity (which is confirmed by the decrease in the content of carbonyl groups in the side chains of proteins compared to the control). ARTICLE INFO Received 4. 1. 2023 Revised 19. 5. 2023 Accepted 2. 6. 2023 Published xx.xx.201x Regular article https://doi.org/10.55251/jmbfs.9771