Problems of Biological Medical and Pharmaceutical Chemistry

Introduction. Phenolic compounds have a wide range of biological activity. They are part of phytopharmaceuticals for the treatment of diseases that are among the most common diseases that require long-term therapy and prevention. In the ethnopharmacology of Siberia, Solidago dahurica Kitag. is used as a means for removing kidney stones, as a diuretic, and as an anti-inflammatory agent. The study of the phenolic compounds of Solidago dahurica is an important task.

The aim of the study – to study the composition and content of phenolic compounds in Solidago dahurica, to assess the dynamics of their accumulation, and to predict the biological activity of the contained phenolic compounds.

Material and methods. The object of the study is the above-ground part of S. dahurica, collected in various phases of vegetation in the Irkutsk region. The isolation and identification of phenolic compounds were carried out using microcolumn chromatography. The content of phenolic compounds was determined using permanganate titration, and the sum of flavones was determined using high-performance liquid chromatography. The content of phenolic compounds was determined by permanganate titration, and the sum of flavonoids and phenolic carboxylic acids was determined by spectrophotometry using a Leki SS1207 spectrophotometer. The prediction of biological activity was carried out using the PASS software, with a threshold probability level of Paі 0.500.

Results. As a result of the conducted studies, 8 compounds were isolated and identified, of which 4 flavonols were quercetin–3-O-rutinoside (rutin), quercetin-3-O-glucoside, isoramnetin-3-O-rutinoside (narcissin), kaempferol-3-O-rutinoside; 3 phenolic carboxylic acids - 3–caffeoylquinic acid (chlorogenic), 3,5-dicofeylchinic and 4,5-dicofeylchinic; as well as bisglucoside – leiocarposide. The dominant phenolic compounds were chlorogenic acid (3,11 mg/g) and 3,5-dicaffeoylquinic acid (2,96 mg/g). In S. dahurica, 5 compounds were discovered for the first time: rutin, narcissin, 3,5- and 4,5-dicaffeoylquinic acids, and leiocarposide. When studying the accumulation dynamics, it was found that the maximum amount of phenolic compounds, as well as flavonoids and phenolic carboxylic acids, accumulates in the aerial part during the flowering period – 8,48%, 4,98%, and 1,44%, respectively. The isolated phenolic compounds have a high predictive level of the following types of biological activity: anti-inflammatory, antioxidant, choleretic, diuretic, and hemostatic (Pa from 0,504 to 0,992).

Conclusions. 8 phenolic compounds – flavonoids, phenolic carboxylic acids and bisglucoside have been isolated and identified from the above-ground part of S. dahurica. The maximum amount of these compounds is accumulated during the flowering period. The isolated phenolic compounds have a high prognostic level in relation to the manifestation of anti-inflammatory, antioxidant, choleretic, diuretic and hemostatic activity.

Introduction. The growing incidence of acute poisoning with fly agarics is becoming a serious medical and social problem. The capsules containing crushed dried raw materials of red fly agaric (Amanita muscaria (L.) Lam.) and/or panther fly agaric (Amanita pantherina (DC.) Krombh.) are most often found on trading platforms. The raw materials contain psychoactive compounds such as muscimol, ibotenic acid and 1-Methyl-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylic acid, which are the reason for their recreational use. The development of methods for the qualitative detection of target substances in capsule extracts is of interest to modern toxicology, since chemical toxicological analysis (CTA) is an essential element of clinical laboratory diagnostics. Obtaining chromatograms and mass spectra of native molecules during capsule analysis will simplify the development of a method for analyzing biological fluids in the future for CTA.

The aim of the study is to develop a method for the qualitative analysis of capsules containing dried raw materials of red and panther mushrooms using liquid tandem chromatography-mass spectrometry.

Material and methods. Red and panther mushroom capsules from the Lesnoy Lekar online store (Russia) were taken as objects of study. Detection and identification of target substances in capsule extracts were carried out on a Nexera XR liquid chromatograph with an LCMS-8050 tandem mass spectrometer (Shimadzu, Japan) using Shim-pack FC-ODS, Shim-pack VP-ODS C8-Phenyl, Shim-pack GIST C18-Aqua HP columns, and a Shimadzu electronic library of mass spectra.

Results. It was found that the optimal method for isolating substances from capsules is methanol extraction and filtration using filter nozzles. Studies have shown that the use of the Shim-pack GIST C18-Aqua HP column and gradient elution mode allows achieving the best parameters for chromatographic separation of muscimol and muscarine.

Conclusions. A method has been developed for isolating muscarine and muscimol from capsules containing crushed dried raw materials of red and panther fly agaric and their qualitative detection using a Nexera XR modular liquid chromatograph with an LCMS-8050 tandem mass spectrometer (Shimadzu, Japan). The Shim-pack GIST C18-Aqua HP column (3 mm × 150 mm, 3 μm) (Shimadzu, Japan) is optimal for analyzing these highly polar hydrophilic toxicants. The following were determined in the methanol extract: muscarine – retention time 3.48 min, muscimol – 2.35 min (the spectrum of ion products had a convergence with the certified library of the device of more than 80%). The obtained data can be of value for clinical and analytical toxicology, and will be further used to develop methods for detecting toxicants in biological objects.

Introduction. Anatomical preparations are not only important auxiliary material in training highly qualified specialists in the natural sciences but also form the basis for scientific research. Most demonstration and educational preparations are wet, that is, they are conserved in preservative liquids and require constant monitoring and periodic restoration. Formalin, despite its well-known carcinogenic, mutagenic and toxic properties, remains the most commonly used preservative solution worldwide. Thus, there has long been a need to search for alternative solutions that also have antiseptic, fixing and preserving properties, but are safe for human health and the environment.

The aim of the study is to conduct a literature review to search for promising options for preservative solutions that do not contain formalin; consider their safety and effect on fixed tissues, study the positive and negative properties.

Material and methods. The review includes data from studies studying various methods of fixing pathological material for the manufacture of wet anatomical preparations. The sources for the review were the PubMed, Google Scholar, ResearchGate, eLibrary.ru, CyberLeninka databases.

Results. This review examines monocomponent preservative solutions based on glyoxal, sodium benzoate, sodium azide, polyhexamethylene guanidine hydrochloride and saturated salt solutions and conducts a comparative analysis of them.

Conclusion. Currently developed alternative preservative solutions have a number of significant drawbacks, which prevents them from becoming a full-fledged replacement for formalin. Further search and improvement of methods for preserving pathological preparations should make it possible in the future to abandon the use of preservative solutions that contain toxic substances that pose a danger to human health and the environment.

Introduction. Improving the effectiveness of cancer treatment remains an urgent task today. The use of various transport forms for therapeutic agents makes it possible to improve their bioavailability and specificity of distribution in the body, reducing toxic effects.

Aim. This study aimed at evaluating the effect of complexes of the native protein apolipoprotein A-I with cytostatic drugs of various concentrations [0.1 and 1 mkg/ml] on the activation of caspase 3/7 in Ehrlich ascites carcinoma cell culture.

Materials and methods. ApoA-I was isolated from the HDL fraction of human blood plasma. The protein was conjugated with cytostatic drugs (actinomycin D, doxorubicin, vinblastine, melphalan). The experiments were carried out on cell culture of Ehrlich's ascites carcinoma. Incubation of Ehrlich ascites carcinoma cells with the studied complexes in various dosages was carried out for 24 hours, 30 minutes before the end the reagent CellEvent^TM Caspase-3/7 Green Detection Reagent (Thermo Fisher Scientific, USA) was added. The calculation of the number of apoptotic cells was recorded using the Program AxioVizion V.4.5.

Results. It has been shown that the addition of ApoA-I complexes with antitumor drugs promotes the activation of caspases 3/7 in Ehrlich ascites carcinoma cells when using cytostatic drugs in minimal concentrations [0.1 µg/ml]. Drugs without a carrier did not cause this effect.

Conclusion. The use of ApoA-I as a transport form of antitumor drugs contributes to a more effective activation of apoptosis in cells of Ehrlich ascites carcinoma in relation to cytostatics without a carrier.

Introduction. Macrolides are a class of broad-spectrum antibiotics that are prescribed for the treatment of various infectious diseases, both local and systemic. Although the use of macrolides is generally considered safe, some patients may be at risk of dose-dependent adverse drug reactions (ADRs), which may reduce the safety of treatment for this category of patients.

The aim of the study was to investigate the effect of CYP3A4 isoenzyme activity on the safety of erythromycin therapy in patients with bacterial complications of influenza.

Material and methods. An open-label, non-comparative, prospective study was conducted in which 40 patients with a confirmed diagnosis of bacterial complications of influenza affecting the lower respiratory tract (ICD-10 codes J10.0-J10.1) who were treated in an infectious diseases hospital, received erythromycin tablets for 5 days. CYP3A4 activity was assessed by high-performance liquid chromatography in relation to the concentrations of the endogenous substrate and its metabolite in urine (6β-hydroxycortisol/cortisol).

Results. and discussion: It was found that the metabolic ratio values on day 1 of the study (corresponding to the baseline level of CYP3A4 isoenzyme activity) and in the dynamics were significantly lower in patients with any recorded NLE. It was also found that the occurrence of nausea was significantly more frequent in patients with low baseline CYP3A4 activity. No statistically significant associations with the metabolic ratio were found for the other individual ADRs.

Conclusions. The study in patients with bacterial complications of influenza demonstrated the influence of CYP3A4 isoenzyme activity on the safety of erythromycin therapy. Thus, determining the level of CYP3A4 activity may allow predicting the development of nausea during erythromycin use.

Introduction. Hyperproduction of reactive oxygen species (ROS) has been implicated in the pathogenesis of several neurological and neurodegenerative diseases. Therefore, antioxidants are able to have a positive effect on the functioning of the central nervous system, preventing the development of oxidative stress. Pyridine is one of the most important heterocyclic nuclei providing antiepileptic, antidepressant, sedative, antitumor and antimicrobial effects. The pyridine ring is also a structural component of the compounds under study, 2,5-diaryl-8,8-dimethyl-3,7,8,9-tetrahydro-2H-pyrido[4,3,2-de]cinnoline-3-ones. Therefore, we were interested to study the effect of these substituted pyridocinnolin-3-ones on the function of the central nervous system through the evaluation of their nootropic activity.

Target. Study of the nootropic activity of 2,5-diaryl-8,8-dimethyl-3,7,8,9-tetrahydro-2H-pyrido[4,3,2-de]cinnolin-3-ones.

Material and methods. 2,5-Diaryl-8,8-dimethyl-3,6,7,8-tetrahydro-2H-pyrido[4,3,2-de]cinnoline-3-ones were selected as the objects of the study. These compounds were prepared by reacting 2-aryl-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydroquinoline-4-carboxylic acids with arylhydrazines. The first stage in the study of nootropic activity was the assessment of exploratory activity in vivo in the «Open Field» model, which allows assessing motor, exploratory activity and stress level. Motor activity is determined by counting the number of intersections of sectors, research - the number of peeks into the holes and lifts on the hind legs, anxiety – the number of fecal boluses and urination. The number of acts of grooming (self-combing) is also recorded.

Results. The structure of the compounds in order to predict biological activity was analyzed in the PASS online program (probability of nootropic activity (p_a) from 61% to 78%). Motor activity in animals during administration of compounds I, III and V exceeds the value of the reference standard (piracetam) by 21–34%. Pronounced stimulation of research behavior is observed under the influence of compounds I and III, with 28% and 54% higher piracetam, respectively, and mexidol in pyridocinnoline I. Activation of indicative reactions exceeding both reference standards (piracetam and mexidol) by an average of 16%, 34% and 68% is characteristic of samples I, III and V. Thus, pyridocinnolines I, III and V, which contain methyl groups, chlorine and fluorine atoms in their structure, have pronounced indicative research activity. The obtained results are confirmed by the prediction data in the PASS program.

Conclusions. The effect of pyridocinnoline-3-ones on exploratory behavior confirms the expediency and prospects of further pharmacological screening, in particular, the study of their effects on cognitive functions.

Introduction. We may use the sources of anthocyanins for prevention of glaucoma in the medicine practice. Pharmacopoeia source of anthocyanins is fruits of bilberry. Related species such as Vaccinium arctostaphyllos may be additional source of anthocyanins along with Vaccinium myrtillus.

Aim – the comparative investigation of anthocyanin profile of Vaccinium arctostaphylos and V. myrtillus fruits and its effects at the progress of glaucoma.

Material and methods. We prepared two types of extracts: without and with addition of hydrochloric acid. We got anthocyanins profile by high-performance liquid chromatography with UV-detector. Glaucoma model was prepared by injection of polystyrene microballons in anterior chamber of rats with subsequent measurement of activity of succinate dehydrogenase and concentration of thiobarbituric acid reactive products (TBARS) in the rat’s brain tissues.

Results. Water-alcohol extracts with addition of hydrochloric acid from bilberry fruits contain three anthocyanins: delphinidin, cyanidin, mixture peonidin and malvidin. Cyanidin is the major components in the both types of fruits. Water-alcohol extracts without hydrochloric acid content nine substances: delphinidin 3-galactoside, delphinidin 3-glucoside, cyanidin 3-galactoside in mixture with delphinidin 3-arabinoside, cyanidin 3-glucoside, petunidin 3-glucoside, peonidin 3-glucoside, malvidin 3-glucoside. Malvidin 3-glucoside and delphinidin 3-glucoside are the main glycosides in the V. arctostaphylos fruits, cyaniding 3-glucoside and glycosides of delphinidin – in the V. myrtillus fruits. Water-alcohol extracts with addition of hydrochloric acid had more effective as neuroprotective agent by increasing concentration of succinate dehydrogenase and decreasing amount of TBARS in the brain.

Conclusion. Extracts from bilberry fruits have neuroprotective effect by presence of total amount of anthocyanins and may be used for prevention of glaucoma.