Category Archives: Uncategorized

1,3-Oxazol-4-ylphosphonium salts as new non-peptide inhibitors of furin

T. V. Osadchuk1, V. K. Kibirev1,2, O. V. Shybyryn1, A. V. Semyroz1,
Ye. S. Velihina1, Е. R. Abdurakhmanova1, V. S. Brovarets1

1V.P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry,
National Academy of Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv

Received: 22 February 2019; Accepted: 17 May 2019

A series of novel triphenylphosphonium derivatives of 1,3-oxazole containing at C2 and C5-positions electron withdrawing or electron-donating groups were synthesized and characterized by 1H, 31P NMR and IR spectroscopy, element analysis and chromato-mass spectrometry. These compounds were found to be a new class of non-peptide inhibitors of furin. Depending on the chemical structure, they inactivated enzyme at micromolar level by mechanism of competitive, non-competitive or mixed inhibition. Evaluation of the synthesized derivatives as furin inhibitors showed that among the triphenylphosphonium salts studied by us, oxazole 12 containing 2,4-dichlorophenyl- in the C2-position and MeS-group at C5 is the most active (Ki = 1.57 μM) competitive inhibitor of furin. Our results provided evidence that chemical modification of 1,3-oxazole-4-yl-triphenylphosphonium salts may be useful for developing new more potent and selective inhibitors of furin.

Born in Ukraine: Nobel prize Winners Ilya Mechnikov, Selman Waksman, Roald Hoffmann AND Georges Charpak

T. V. Danylova1, S. V. Komisarenko2

1National University of Life and Environmental Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Received: 18 February 2019; Accepted: 14 March 2019

Our country has not yet gained recognition from a Nobel Committee, however, some Nobel Prize winners were born in the territory, which belongs to present-day Ukraine. Among them are the father of innate cellular immunity Ilya Mechnikov; the famous microbiologist and biochemist Selman Waksman, whose studies had led to the discovery of streptomycin; the outstanding chemist, poet and playwright Roald Hoffmann, and the prominent physicist Georges Charpak who invented and developed particle detectors, in particular, the multiwire proportional chamber. This paper aims to outline briefly the main stages of their scientific activity.

Development on knowledge of hormone biochemistry in the works of the Nobel prize laureates of the first half of the 20th century: F. G. Banting, John J. R. Macleod, H. O. Wieland, A. O. Windaus, A. F. Butenandt, L. Ružička, E.Kendall, P. Hench, T. Reichstein

R. P. Vynogradova, V. M. Danilova, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Received: 18 February 2019; Accepted: 14 March 2019

The first half of the 20th century was marked by significant scientific advances in the study of hormones and vitamins. Among the first resear­chers working with hormones were F. Banting and J. Macleod, who discovered and characterized the pancreatic hormone insulin. This discovery catalyzed advances in the understanding of the mechanisms regulating biochemical processes – a new topic in the field of biological chemistry. The next important stage in the development of knowledge on biologically active substances was the works of organic chemists G. Wieland, A. Windaus, A. Butenandt and L. Ružička. They almost simultaneously identified and characterized the chemical structures of bile acids, vitamin D as well as female and male sex hormones. They found that all of these compounds are of a steroid nature and identified cholesterol as the starting material for their synthesis in the body. The studies of highly-active substances of steroid nature were continued by E. Kendall, F. Hench and T. Reichstein. They synthesized and investigated the structure and biological effects of corticosteroids, the hormones produced in the adrenal cortex. They were first to develop a method for the commercial manufacturing of cortisone, a hormone which is widely used to treat inflammatory processes. Thus, in the first half of the 20th century, organic chemists gave biochemists knowledge on the structure of essential for the human body substances – steroid compounds.

Lipid profile parameters and oxidative processes intensity in the persons who have been affected by low doses of radiation

V. L. Sokolenko, S. V. Sokolenko

Bohdan Khmelnytsky National University of Cherkasy, Ukraine;

Received: 10 October 2018; Accepted: 14 March 2019

The aim of the work was to analyze the relationship between the main parameters of lipid metabolism and the intensity of oxidative processes among the inhabitants of the territories contaminated by radionuclides as a result of the Chornobyl accident. We examined 50 persons from the control group and 50 persons from the territories of strengthened radioecological control (density of soil contamination by isotopes 137Cs 3.7∙104–18.5∙104 Bq/m2, 50 persons). All examined were the students aged 18 to 24, who had no acute illnesses during the study. We determined the parameters of lipid metabolism, oxidative processes and antioxidant system. A positive correlation of all analyzed lipid metabolism parameters (except for HDL-C) with MDA, ceruloplasmin and the index of oxidative stress was discovered. The highest values of correlation coefficients with oxidative stress indices were observed for low-density lipoprotein cholesterol. Under the conditions of additional emotional stress, the correlation coefficients between the main lipid metabolism and the intensity of oxidative processes increased. Persons who lived for a long time in areas contaminated with radionuclides form the risk group for the development and progression of inflammatory processes. The risk increases under the influence of additional factors of a stressful nature.

New monoclonal antibodies to the Chlamydia trachomatis main outer membrane protein and their immunobiological properties

O. Yu. Galkin1,2, O. B. Besarab1, Yu. V. Gorshunov1, O. M. Ivanova3

1National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”;
2Propharma Plant Ltd., Kyiv;
3Xema Ltd., Kyiv

Received: 18 July 2018; Accepted: 14 March 2019

One of the methods that have been widely used in the diagnosis of urogenital chlamydia is an enzyme-linked immunosorbent assay (ELISA), the use of which allows for differential diagnosis. In order to increase the efficiency of ELISA test kits production, for the kits for the diagnosis of urogenital chlamydia, based on the principle of indirect modification, following synthetic positive controls (PCs) can be used: a conjugate of IgM (IgA) normal immunoglobulins and monoclonal antibodies (McAbs) to C. trachomatis major outer membrane protein (MOMP). The goal of this work was to obtain high active and affinity McAbs to the C. trachomatis MOMP as well as the study of its immunobiological properties which are important for future biochemical approaches. The study was conducted using: polyclonal antibodies (PcAbs) to C. trachomatis; recombinant major outer membrane protein (MOMP) (191-354 a.r.; W4-W5); epitope mapping based on phage display technology. The original set from 16 clones of hybridomas, producers of McAbs to the C. trachomatis MOMP has been obtained. More than half of the tested McAbs (8 out of 14) were characterized by a rather high titer (≥1:800), and three of them had a titer of ≥1:1600. In general, the McAbs titer was correlated with the value of the affinity constant: McAbs with higher titles were characterized by a high value of the affinity constant. For McAbs with a titer of <1:800, the average Ka is 5.2×109 M-1, while for McAbs with a titer ≥1:800 – Ka = 10.7×109 M-1. Antigenic determinants of two McAbs 293F4 and 291F8 that actively competed with PcAbs are represented by two linear sequences of 320-325 a.r. and 326-330 a.r., respectively. The epitope, which interacts with McAb 296G2, is represented by a linear sequence of 347-352 a.r. McAb 296G2 did not show active competition with serum PcAbs. The resulting set of data allows selecting McAbs for use in PCs of the ELISA kit for the detection of IgA or IgM antibodies to C. trachomatis.

New anti-candida active nitrogen-containing bisphosphonates as inhibitors of farnesyl pyrophosphate synthase Candida albicans

L. O. Metelytsia, D. M. Hodyna, O. L. Kobzar,
V. V. Kovalishyn, I. V. Semenyuta

V. P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry,
National Academy of Sciences of Ukraine, Kyiv;

Received: 05 February 2019; Accepted: 14 March 2019

In our previous work, a number of new nitrogen-containing bisphosphonates (N-BPs) with high predicted and experimental antifungal activity were presented as potential Candida albicans farnesyl pyrophos­phate synthase (FPPS) inhibitors. To confirm this hypothesis, a homologous C. albicans FPPS model with high-quality scores has been developed and used in present work to study the molecular mechanism of nit­rogen-containing bisphosphonates action as anti-Candida agents. The known FPPS inhibitors ammonium 2-(Pyridin-2-ylamino)ethylidene-1,1-bisphosphonate, risedronate and alendronate were used in molecular docking analysis. The molecular docking analysis of the new N-BPs demonstrated a number of common features of all ligand’s interaction in the active center of FPPS C. albicans. It is established that the ligands phosphonate groups are the key elements in the formation of the stable ligand-protein complexes with binding energy in a range (ΔG) from –6.6 to –7.1 kcal/mol due to a significant number of electrostatic, hydrogen and metal-acceptor bonds. It is confirmed that the new studied N-BPs 1 and 3 with high anti-Candida activity are FPPS inhibitors.

Insulin resistance in obese adolescents and adult men modifies the expression of proliferation related genes

O. H. Minchenko1, Y. M. Viletska1, D. O. Minchenko1,2, V. V. Davydov3

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Bohomolets National Medical University, Kyiv, Ukraine
3SI “Institute of Children and Adolescent Health Care,
National Academy of Medical Sciences of Ukraine”, Kharkiv

Received: 11 December 2018; Accepted: 14 March 2019

Numerous data demonstrate that key regulatory factors, enzymes and receptors including HSPA5, MEST, SLC1A3, PDGFC, and ADM represent poly-functional, endoplasmic reticulum stress-dependent proteins, which control variable metabolic pathways. The expression level of genes of these proteins in the blood and subcutaneous adipose tissue of obese adolescents and adult men with and without insulin resistance was studied. It was shown that in blood of obese adolescents without insulin resistance the expression level of SLC1A3, HSPA5, MEST, and PDGFC genes was significantly increased, but development of insulin resis­tance led to down-regulation of these genes expression except HSPA5 gene as compared to the control group as well as to the group of obese adolescents without insulin resistance. At the same time, the expression level of ADM gene did not change significantly in obese adolescents without insulin resistance, but the development of insulin resistance led to down-regulation of this gene expression. In subcutaneous adipose tissue of obese adult men without insulin resistance the level of SLC1A3 gene expression was decreased, although ADM, MEST, and HSPA5 genes – increased. It was also shown that the development of insulin resistance in obese men affected the expression level of ADM and SLC1A3 genes only. Results of this investigation provide evidence that insulin resistance in obese adolescents and adult men is associated with specific changes in the expression of genes, which related to proliferation and development of obesity and insulin resistance as well as to endoplasmic reticulum stress and contribute to the development of obesity complications.

Kinetic properties of Na(+),K(+)-АТРase of spermatozoa from fertile and infertile men under effect of calix[4]arene C-107

R. V. Fafula, O. I. Meskalo, A. S. Besedina,
Io. A. Nakonechnyi, D. Z. Vorobets, Z. D. Vorobets

Danylo Halytsky Lviv National Medical University, Ukraine;

Received: 12 November 2018; Accepted: 14 March 2019

The calix[4]arene C-107 (5,17-diamino(2-pyridyl)methylphosphono-11,23-di-tret-butyl-26,28-dihydro­xy-25,27-dipropoxy-calix[4]arene) effects on the kinetic properties of Na+,K+-ATPase in spermatozoa of fertile (normozoospermia) and infertility men (oligozoospermia, and asthenozoospermia) were studied. It was shown that in spermatozoa of healthy men calix[4]arene С-107 inhibited Na+,K+-ATPase activity and decreased the maximum reaction rate of ATP hydrolase reaction without affecting the coefficient of (half-) activation by ATP and Hill coefficient nH. In оligo- and asthenozoospermic samples of spermatozoa almost a 2-fold decrease of cooperativity coefficient nH of ATPase inhibition with calyx[4]aren C-107 was observed. In normozoospermic samples of spermatozoa the KMgCl2  for Na+,K+-ATPase was decreased at calix[4]arene C-107 high concentrations (≥50 nM) in the incubation medium in contrast to oligozoospermic samples of spermatozoa where KMgCl2 was increased only at high calix[4]arene C-107 concentration (100 nM). The increase of the KMgCl2 in the entire range of investigated calix[4]arene concentrations and the decrease of cooperativity coefficient nH of MgCl2 activating effect were detected in asthenozoospermic samples of Na+­,K+-ATPase.

Сalix[4]arene chalcone amides effects on myometrium mitochondria

S. G. Shlykov1, A. M. Kushnarova-Vakal1, A. V. Sylenko1,
L. G. Babich1, О. Yu. Chunikhin1, O. A. Yesypenko2,
V. I. Kalchenko2, S. O. Kosterin1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv

Received: 19 November 2018; Accepted: 14 March 2019

Mitochondria are a key player in a wide range of the most important functions of the cell. Calixarenes are supramolecular compounds that have been widely used in bioorganic chemistry and biochemistry. The aim of this work was to study the effects of calix[4]arenes with two (С-1012, С-1021), three (С-1023, С-1024) and four (С-1011) chalcone amide groups on the myometrial mitochondria membranes polarization, Ca2+ concentration in the matrix of these organelles ([Ca2+]m ) and on the average hydrodynamic diameter of mitochondria. It was shown that permeabilized myometrium cells incubation with calix[4]arenes containing two or more chalcone amide groups, was accompanied by an increased level of myometrial mitochondria membranes polarization. All studied calix[4]arenes increased [Ca2+]m values in the absence and in the presence of exogenous Ca2+. The values of [Ca2+]m in the absence of exogenous Ca2+ were higher at mitochondria incubation in Mg2+-containing, than in Mg2+,ATP-containing medium. Incubation of isolated mitochondria with the studied calix[4]arenes resulted in changes of mitochondria volume: at incubation with С-1012, С-1021, C-1023 the average hydrodynamic diameter was decreased, while with С-1011 it was increased. Thus, we have shown that a short-term (5 min) incubation of mitochondria in the presence of 10 µM calix[4]arenes, which contain from two to four chalcone amide groups, increased the level of mitochondria membranes polarization, ionized Ca concentration in the matrix and had different effects on the mitochondrial volume.

Adaptive respiratory response of rat pancreatic acinar cells to mitochondrial membrane depolarization

B. O. Manko, O. O. Bilonoha, V. V. Manko

Ivan Franko National University of Lviv, Ukraine;

Received: 06 December 2018; Accepted: 14 March 2019

The dependence of uncoupled respiratory capacity of intact pancreatic acini on oxidative substrate supply and functional cell state has not yet been studied in detail. In this study, the respiratory responses of isolated pancreatic acini to FCCP were measured with Clark electrode and mitochondrial membrane potential was assessed with rhodamine123 fluorescence. The response of acini to FCCP was characteri­zed with maximal uncoupled respiration rate, optimal FCCP concentration, respiration acceleration and decele­ration. Maximal uncoupled respiration rate substantially increased upon the oxidation of glucose + glutamine (3.03 ± 0.54 r.u.), glucose + glutamine + pyruvate (2.82 ± 0.51 r.u.), glucose + isocitrate (2.71 ± 0.33 r.u.), glucose + malate (2.75 ± 0.38 r.u.), glucose + monomethyl-succinate (2.64 ± 0.42 r.u.) or glucose + dimethyl-α-ketoglutarate (2.36 ± 0.33 r.u.) comparing to glucose alone (1.73–2.02 r.u.) or no substrate (1.76 ± 0.33 r.u.). The optimal FCCP concentration was the highest (1.75 μM) upon glucose + glutamine + pyruvate combination and the lowest (0.5 μM) upon glutamate, combinations of glucose with isocitrate, malate, succinate or α-ketoglutarate. Respiration acceleration after FCCP application was the highest with dimethyl-α-ketoglutarate. Following the peak respiration, time-dependent deceleration was observed. It increased with FCCP concentration and depended on oxidative substrate type. Deceleration was the highest upon malate or isocitrate oxidation but was not observed in case of glutamine or dimethyl-α-ketoglutarate oxidation. Pyruvate alone or in combination with glutamine and glucose significantly decreased the depolarizing effect of FCCP on mitochondrial membrane potential and increased respiration elasticity coefficient with respect to the membrane potential change. Thus, in pancreatic acinar cells, the combination of pyruvate, glutamine and glucose enables the optimal adaptive respiratory response to membrane depolarization.