Problems of Biological Medical and Pharmaceutical Chemistry
Peer-review scientific and practical journal
Editor-in-chief
- Nikolay I. Sidelniko, Doctor of Agricultural Science, the academician of RAS
Publisher
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Publishing House «Russkiy Vrach»
Founder
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All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
About
The journal publishes materials on biological, medical, and pharmaceutical chemistry, directly related to problems of modern medicine. Established in 1998.
Sections
- Pharmaceutical chemistry
- Biological chemistry
- Medical chemistry
- Problems of experimental biology and medicine
- Bioelementology
- Plant protection and biotechnology
- Brief reports
Current Issue



Vol 28, No 4 (2025)
Pharmaceutical chemistry
Computer simulation of the processes of complexation between new derivatives of quinazoline-4(3H)-one, quinazoline-2,4(1H,3H)-dione and cytochrome P-450 sterol-7Α-hydroxylase (CYP27A1) when studying their antimicrobial effects against Klebsiella pneumoniae
Abstract
Introduction. Multi-resistance of Klebsiella pneumoniae to known antimicrobial agents determines the need for urgent development of new drugs exhibiting an antibacterial effect. Compounds used in the treatment of non-infectious pathologies, affecting various protein targets and realizing the "antimicrobial potential", seem promising in the search for new derivatives. Of particular interest is the predicted high probability of inhibition of sterol-27-hydroxylase cytochrome P-450 (CYP27A1) by new derivatives of quinazoline-4(3H)-one and quinazoline-2,4(1H,3H)-dione with the possibility of preventing the processes of conversion of cholesterol, as the main component of macrophage lipid rafts on which pathogen adhesion occurs. Activation of phagocytosis, as well as disruption of the formation of the polysaccharide capsule of K. pneumoniae, is considered one of the probable mechanisms of the antibacterial effect of the studied substances.
The aim – determination of reactivity parameters and potential pharmacological activity of quinazolin-4(3H)-one and quinazolin-2,4(1H,3H)-dione derivatives; prediction of the mechanism of antimicrobial action of new quinazolinones against K. pneumoniae based on the formation of a complex with CYP27A1.
Material and methods. The probability of CYP27A1 inhibition by new quinazolinone derivatives was predicted using the PASS program. Quantum chemical parameters were calculated using the parameterized PM7 method in the MOPAC 2016 program. The structural characteristics of the studied substances were determined using ProTox 3.0. Some pharmacokinetic parameters were assessed using the admetSAR software tool. A three-dimensional model of the protein structure of CYP27A1 was generated on the AlphaFold platform; optimal conformation and further clustering of protein sequences using the MMseqs2 and Foldseek algorithms. The determination of the energy parameters of the intermolecular complexes “quinazolinone CYP27A1”, as well as the identification of the amino acid sites to which the quinzolinone derivative binds, was carried out using the Swiss Dock system.
Results. Quantum-chemical and structural parameters of quinazoline-4(3H)-one and quinazoline-2,4(1H,3H)-dione derivatives, as well as their intermolecular complexes with CYP27A1, were established, and some pharmacokinetic parameters of the studied substances were predicted.
Conclusions. Control of the interaction of the compound with the lipid raft of the macrophage and an increase in the degree of penetration of the pathogen into its cell can be considered as a probable mechanism of the antibacterial action of quinazoline-4(3H)-one and quinazoline-2,4(1H,3H)-dione derivatives against Klebsiella pneumoniae. An increase in the number of naphthyl radicals in their molecule leads to a decrease in reactivity. Naphthyl radicals do not act as a pharmacophore of antimicrobial action against K. pneumoniae.



Biological activity of deep eutectic solvent-based extracts of Chamaeneríon angustifolium (L.) Scop.
Abstract
Introductioin. For the use of plant extracts in pharmaceutical, cosmetic and food industries it is important to study their biological activity towards cells and bacterial structures, as well as their antioxidant properties.
Aim of the study – a comprehensive evaluation of the biological properties of plant extracts of leaves of Ivan-tea narrow-leaved (Chamaeneríon angustifolium (L.) Scop.) based on deep eutectic solvents (DESs), compositions [ChCl][Gly]2[H2O]11, [ChCl][Cit]2[H2O]43 and [Cit][Gly]4[H2O]20, in comparison with pure solvents.
Material and Methods. The ability of the obtained extracts to inhibit ABTS and DPPH free radicals was determined. The antimicrobial activity of the extracts and extractants against bacterial cultures of Bacillus pumilus, Microbacterium paraoxydants and Acinetobacter lwoffii and the cytotoxicity of DES and plant extracts against HeLa cervical carcinoma cell line and HEK293 human embryonic kidney cell line were determined using MTT assay.
Results. It was shown that [ChCl][Gly]2[H2O]11 based extract inhibited DPPH more strongly than ABTS, while on the contrary [ChCl][Cit]2[H2O]43 and [Cit][Gly]4[H2O]20 based extracts inhibited ABTS radical more intensely. It was also found that all the DES based extracts of C. angustifolium at high concentrations (10, 1 vol.%) had pronounced antimicrobial activity against the bacterial strains used. The [Cit][Gly]4[H2O]20 based extract was found to be the most toxic to all bacterial cultures. The cytotoxic effect was less pronounced for healthy cells with [ChCl][Cit]2[H2O]43 and for cancer cells with [ChCl][Gly]2[H2O]11.
Conclusions. The results of the work are a scientific basis for further development of methods of application of DESs and extracts based on them in food and pharmaceutical industries.



Comparative study of extraction methods from biological material of tricyclic antidepressants: imipramine, clomipramine, amitriptyline
Abstract
Introduction. Tricyclic antidepressants (TCAs) are historically the first group of medications used to treat depression. They work by affecting the duration of monoamines presents in the presynaptic cleft in the brain and blocking certain receptors, such as m-cholinergic and alpha-adrenergic. TCAs can cause a toxicological effect and lead to fatal poisoning. This group of drugs is often used among drug addicts, as well as in the criminal environment.
The purpose of the study. Determination of an optimal and effective method for TCAs extraction from biological material using solid-phase (SPE) and liquid-liquid extraction (LLE) for further chemical-toxicological studies.
Materials and methods. The objects of the study were representatives of the TCAs group: imipramine, amitriptyline and clomipramine. Urine was used as a biological material. In the study of SPE method columns produced by Agilent Technologies company were used. These columns contained Evidex and ODS-C18 sorbents. Analutes isolation using LLE were realised according to the one of the most common methods. Analisys were conducted on gas-chromatography with a mass-selective detector (GC/MS).
Results. The method of extraction on a solid sorbent was developed using phosphate buffer with pH 4.8. After using the LLE method, samples with high contamination of co-extracted impurities of biological origin were observed. The error in the results of TCAs extraction using SPE was less than 10%, while the error for the LLE method was 25-35%. A comparison of two different extraction methods was studied using various TCAs concentrations. The advantages of using the ODS-C18 as a sorbent for SPE were determined. Calibration curves were constructed for samples using the SPE method. Limits of detection and quantification for all TCAs were determined.
Conclusions. The advantages of isolating TCAs using the SPE method over the LLE method were revealed.



Development of a technique for quantitative determination of silver by extraction-photometric method in a polyacrylamide-based medical device
Abstract
Introduction. One of the promising directions of using polymer hydrogels in medicine is the production of synovial fluid endoprostheses. Thus, cross-linked polyacrylamide serves as the basis for the medical device “Sterile Silver Ion Waterborne Biopolymer Material”, which is used for intra-articular injections in the treatment of osteoarthritis. The addition of silver ions to polymeric hydrogels is due to its broad spectrum of antimicrobial activity and is used to prevent microbial contamination. However, in order to safely use a medical device containing silver ions and avoid the risk of heavy metal poisoning, the content of this component must be monitored. Among the various methods of quantitative analysis of metals, the extraction-photometric method is the easiest to reproduce and does not require expensive equipment, which allows to control the content of the element in low content.
Purpose of the study. Development of a technique for quantitative determination of silver in silver-containing medical device based on polyacrylamide hydrogel by extraction-photometric method.
Material and Methods. A sample of medical product “Material-biopolymer water-containing with silver ions sterile”, produced by LLC “NC ‘BIOFORM’, Russia, was used as an object of the study. The silver content in the studied object was determined by the extraction-photometric method.
Results. The technique of quantitative determination of silver in the analyzed medical device by the extraction-photometric method based on the extraction of pre-ionized silver in an organic solvent (dithizone solution in 1-butanol 0.0025 %) through the formation of a high-affinity complex in acidic medium was developed. The absence of placebo (polyacrylamide) influence on determination of silver ions was proved.
Conclusions. This study demonstrated the possibility of silver ions determination in the medical device “Material-biopolymer water-containing with silver ions sterile” after extraction with dithizone solution in 1-butanol 0.0025 % by extraction-photometric method.
It was found that the developed method is applicable in the range of measured concentrations of silver ions from 0.2 to 5.0 μg/mL, the average value of the method opening on 6 levels of silver content is 101.5 %.



Medical chemistry
Analysis of the overall survival of patients with primary glioblastoma of the brain depending on glial fibrillary acidic protein content in blood serum
Abstract
Introduction. Glioblastoma is one of the most aggressive malignant brain tumors characterized by low sensitivity to drug therapy, frequent recurrence after surgical removal of the tumor, and an extremely unfavorable prognosis.
Aim. To analyze the overall survival of patients with primary glioblastoma of the brain based on the glial fibrillary acidic protein (GFAP) content of in blood serum.
Material and methods. 176 patients with primary malignancy grade IV glioblastoma confirmed by morphological and molecular genetic studies (99 men and 77 women) aged 18–82 years who underwent examination and treatment at the N.N. Blokhin National Medical Research Center of Oncology in the period from 2013 to 2024 were enclosed in the study. GFAP concentration in the pre-treatment blood serum was determined by enzyme immunoassay using Human GFAP ELISA kit (Biovendor, Czech Republic). The obtained data were processed using Statistica 10 (StatSoft) and SPSS (IBM) programs. Overall survival analysis was performed using the Kaplan–Meier method with the log-rank test for comparisons. Differences were considered statistically significant at p < 0.05.
Results. 72 of 176 patients were alive at the end of the study, the median follow-up period being 7.8 months, while 104 patients with died within a period of 1 to 103 months the median survival being 9.1 months. The median lifespan of all patients with glioblastoma was 14.8 months. 1-year overall survival rate comprised 56.7±4.2%, 2-year – 39.2±4.3%, 3-year – 30.0±4.1%, 4and 5-year – 26.8±4.0%, 10-year – 15.0±4.4%. The risk of death in patients with primary glioblastoma of the brain was the greatest in the first two years of follow-up from the time of diagnosis. No associations were found with patients’ age and gender, or the side of the tumor lesion. To study the relationship between long-term treatment results and serum GFAP concentrations patients were divided into two groups: 1) patients in whom GFAP was not detected (GFAP-negative) and 2) those with detectable GFAP concentrations (GFAP-positive). Median survival time of 46 GFAP-negative glioblastoma patients was 24.8 months, while in 130 GFAP-positive patients it comprised only 13.7 months (p = 0.047). No differences in the overall survival in GFAP-positive glioblastoma group depending on the serum GFAP concentration were found. The differences in the overall survival depending on the presence or absence of GFAP in blood serum were mostly pronounced in the female group and in patients below 50 years of age. Thus, median survival of GFAP-negative glioblastoma patients below 50 years was several fold higher than in GFAP-positive patients of this age group (70.8 and 12.4 months respectively).
Conclusion. The overall survival of patients with grade IV primary glioblastoma is significantly higher in GFAP-negative than in GFAP-positive group. Hence, the presence of GFAP in blood serum can be regarded as an unfavorable prognostic factor for this disease.



Redox regulation of apoptosis and proliferation in psoriasis patients using omega-3-PUFA-containing drugs by thiol-disulfide exchange
Abstract
Introduction. The relevance of the topic is due to the wide prevalence of psoriasis, which is characterized as increase in the intensity of keratinocyte proliferation. One of the factors contributing to the initiation of proliferation processes is the overreaction of the antioxidant system to inflammation, accompanied by a decrease in the intensity of free radical oxidation.
Material and Methods. The study included patients with an established diagnosis of psoriasis vulgaris, divided into three groups of 15 people, who along with standard therapy received the biologically active supplement "Omega-3 35%". The control group consisted of 15 healthy people. The antioxidant status was assessed by the state of the enzymatic and non-enzymatic link: the activity of catalase and superoxide dismutase of blood hemolysate and their ratio, the concentration of total HS-groups in plasma; the content of end products of lipid peroxidation – by the concentration of malondialdehyde in plasma. The state of apoptosis was determined by flow cytometry, the intensity of proliferation – by clinical manifestations and the PASI index.
Purpose of the work – to study the effect of omega-3 polyunsaturated fatty acids in different doses.
Results. It has been shown that in patients with psoriasis, when added to the standard therapy "Omega-3 35%" at a dose of 3450 mg/day, divided into three doses of 1150 mg at three times a day, an antioxidant effect is manifested, instead of the expected pro-oxidant clinical improvement is insignificant (the decrease in the PASI index is less than 25%), and at a dose of 2640 mg/day once a day for a month, a moderate pro-oxidant effect is observed, which is necessary to reduce excess proliferation, and increase in the intensity of apoptosis, these changes are accompanied by clinical improvement and a decrease in the PASI index by 37% compared to the baseline.
Conclusions. It has been shown that in patients with psoriasis, a shift in redox equilibrium towards oxidized equivalents is necessary to obtain positive clinical dynamics. It is assumed that redox regulation with the addition of omega-3 PUFAs occurs due to a short-term increase in oxidative processes and is realized through thiold-disulfide exchange.



Soluble forms of PD-1 receptor and PD-L1 ligand in patients with renal cell carcinoma
Abstract
Introduction. The study of soluble forms of the receptor and immune checkpoint ligand (ICP) PD-1/PD-L1 is considered one of the promising areas in renal cell carcinoma (RCC). This system is actively involved in the regulation of antitumor immunity and includes the programmed cell death protein PD-1 (programmed cell death protein 1) and two ligands PD-L1, PD-L2. It is known that activation of the PD-1/PD-L1 system simultaneously stimulates apoptosis of antigen-specific T cells and suppresses apoptosis of regulatory suppressor T cells, which allows tumors to "escape" the host immune system.
The aim of the work is to analyze serum levels of sPD-1 and sPD-L1 in patients with renal cell carcinoma, their relationship with clinical and morphological characteristics of the disease.
Material and methods. We studied the levels of sPD-1 and sPD-L1 in the blood serum before treatment in 117 patients with kidney tumors, including 105 patients with renal cell carcinoma (RCC) (63 men and 42 women) aged 33 to 81 years (median 60 years) at various stages of the disease. Benign renal tumor (DOP) was detected in 12 patients (3 men, 9 women) aged 29 to 84 years (median 48.5 years). The control group consisted of 67 healthy individuals (27 men, 40 women) aged 22 to 82 years (median 53 years). The concentration of sPD-L1 and sPD-1 was determined in the blood serum before treatment using the Human PD-L1 Platinum ELISA and Human PD-1 ELISA kits (Affimetrix, eBioscience, USA). Statistical analysis of the distributions of the studied features was performed using the Kolmogorov-Smirnov goodness-of-fit test. To identify differences between values, single-factor and multifactorial variance analysis were used. All calculations were performed on a personal computer using the statistical software package "Statistica for Windows" and SPSS.
Results. A significantly higher median sPD-1 concentration was found in patients with RCC compared to the control; in patients with DOP, the median marker did not differ from the control. The median serum sPD-L1 content in RCC was significantly higher than in the control and did not differ from the group of patients with DOP. Analysis of variance of sPD-1, sPD-L1, sPD-1/sPD-L1 in the blood serum in the control and in patients with RCC did not show a relationship with age. The sPD-L1 index in RCC patients was significantly higher in men than in women. The T index reflected the sPD-L1 index and its concentrations were significantly higher in RCC patients with metastases in regional lymph nodes. The M index in RCC patients reflected serum sPD-L1 levels to the greatest extent. The lowest receptor median was found in clear cell RCC. ROC analysis indicates the prospects for further studies of sPD-L1 as a biomarker in the diagnosis of RCC.
Conclusions. The medians of sPD-1 and sPD-L1 are significantly higher in patients with RCC than in the control. Analysis of variance of the values of the studied parameters in the control and in patients with RCC did not show a relationship with age. The T and M index are closely related to sPD-L levels in RCC. In patients with RCC, significant differences in PD-1 receptor levels were found between different morphological variants of the tumor. Analysis of the diagnostic significance of sPD-L1 in patients with RCC indicates the prospect of further research of the biomarker in the diagnosis of RCC.



Biological chemistry
Comparative analysis of methods for inducing steatosis using a complex of fatty acid bovine serum albumin in an in vitro model on HepG2 cells
Abstract
Introduction. Non-alcoholic fatty liver disease (NAFLD) is a major public health issue characterized by a rapidly increasing prevalence worldwide. NAFLD is associated with excessive lipid accumulation and the development of inflammation in the liver. To study the pathogenic mechanisms of the disease, a steatosis model using immortalized cell lines is widely employed.
The aim of this study was to improve the in vitro steatosis model using HepG2 cells by conducting a comparative analysis of two methods of steatosis induction, utilizing a fatty acid (FA) complex with bovine serum albumin (BSA) prepared by simple dissolution or conjugation.
Material and methods. Cell viability was assessed using the XTT assay. Lipid accumulation in HepG2 cells was measured by the GPO-PAP method, with triglyceride (TG) levels normalized to cellular protein content. Production of the pro-inflammatory cytokine IL-8, a marker of inflammation, was quantitatively determined using an enzyme-linked immunosorbent assay (ELISA). Statistical significance was evaluated using Student's t-test, with p-values < 0.05 considered statistically significant.
Results. The optimal FA concentration that promoted lipid accumulation and inflammation without a marked cytotoxic effect in HepG2 cells was 0.75 mM. The conjugated FA-BSA complex led to a higher TG accumulation compared to the FA-BSA complex prepared by dissolution. However, IL-8 levels were significantly lower in the culture medium of HepG2 cells treated with the conjugated complex compared to those treated with the FA-BSA complex prepared by dissolution.
Conclusions. The use of the conjugated FA-BSA complex allowed for the development of an improved in vitro steatosis model that more closely resembles the physiological mechanisms of NAFLD progression.


