Russian Journal of Bioorganic Chemistry最新文献

Objective: Dehydroabietic acid (DHAA) is an important natural triterpene resin acid with a wide range of pharmacological activities, including antibacterial, antifungal, antiviral, and anti-inflammatory effects, particularly in antitumor activity. The aim of this study was to synthesize a series of novel dehydroabietic acid 1,2,3-triazole derivatives and evaluate their antitumor activity. Methods: Click chemistry was employed to conjugate dehydroabietic acid (DHAA) with 1,2,3-triazole, yielding 12 DHAA derivatives containing 1,2,3-triazole with yields ranging from 64.5 to 90%. The structures of the target compounds were confirmed by ¹H, ¹³C NMR, and HR-MS (ESI). The initial antiproliferative activity of these compounds was assessed in vitro using two tumor cell lines (human cervical cancer cells, HeLa, and human breast carcinoma cells, MCF-7), with cisplatin used as a reference. Results and Discussion: The results indicated that three compounds, VII, VIII, and IX, exhibited significant antiproliferative activity against the tumor cell lines. Among them, compound VIII (4-(4-fluorophenyl)-1-(((1R,4aS)-7-isopropyl-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl)methyl)-1H-1,2,3-triazole) demonstrated particularly remarkable activity, with an IC₅₀ value of 22.1 μM against HeLa cells and 23.2 μM against MCF-7 cells. Conclusions: This study provides useful strategies for the design and synthesis of novel antitumor agents. The synthesized compounds exhibit some inhibitory effects on tumor cell growth and proliferation while causing minimal damage to normal cells, warranting further investigation.

Objective: To characterize the antimicrobial resistance gene (ARG) profiles of the WHO 2024 N. gonorrhoeae reference strains and compare the CARD/RGI and NG-STAR genotyping methods. Methods: We performed in silico analysis of 29 reference genomes using the Resistance Gene Identifier (RGI) tool and the Comprehensive Antibiotic Resistance Database (CARD), benchmarking results against the NG-STAR scheme. Results and Discussion: β-Lactam and macrolide resistance genes were most prevalent. Key determinants included mtrA, penA, rpsJ, and mtrC. Strains WHO_Q and WHO_beta harbored the most ARGs (10 each). Concordance between CARD/RGI and NG-STAR was 39.4%, with CARD providing broader mechanistic coverage and NG-STAR offering superior allele-level resolution. Conclusions: The study provides a benchmark for genomic AMR prediction, demonstrating the complementary value of general-purpose and species-specific typing schemes for surveillance.

Objective: Diabetes and diabetes-induced diseases, such as cataracts, are growing global health problems, with ongoing research into better treatment options. In India, the prevalence of blindness is 15 per 1000 people, with cataracts alone accounting for 80% of cases. In this context, chalcones are frequently investigated by scientists as potential agents with diverse biological activities. Results and Discussion: Compounds IIa and IIc, which demonstrated ALR2 inhibition in in silico studies, were evaluated for the inhibition of rat lens aldose reductase, with Epalrestat used as the standard drug. Both compounds showed promising results, with IC₅₀ values lower than that of the standard drug. Compounds IIb and IId, which exhibited promising results as PPAR-γ agonists in in silico analysis, were further evaluated in vivo using streptozotocin (STZ) and high-fat diet (HFD)-induced diabetic mice. The disease control group showed mature cataracts, while the treated groups exhibited the opposite at the end of the study. The mice were assessed for various parameters, including blood glucose, serum total cholesterol, triglycerides, HDL, LDL, aldose reductase levels, antioxidant enzyme activity, and lipid peroxidation at the conclusion of the treatment, in comparison with standard drug-treated groups (Epalrestat and Pioglitazone). Levels of aldose reductase, blood glucose, triglycerides, cholesterol, and LDL in the lens were significantly decreased, whereas antioxidant enzymes, total proteins, soluble proteins, and HDL were significantly increased in the treatment groups. The higher dose (200 mg/kg) of compound IIb showed pronounced protection. Six chalcones were synthesized via the Claisen–Schmidt condensation reaction and evaluated for anti-diabetic activity in silico against ALR2 and PPAR-γ receptors. The promising compounds were then assessed for their relative antidiabetic activity in vitro and in vivo. Conclusions: The results suggest that compound IIb may be effective against hyperglycemia-induced activation of the polyol pathway, oxidative and osmotic stress, as well as the subsequent development of diabetic cataracts.

Objective: This study aimed to improve the DNA-nanomachine (DNM) platform based on the 10– 23 RNA-cleaving DNAzyme for efficient recognition of dsDNA at near-physiological temperatures. Two DNM variants with distinct scaffold architectures were designed and compared with the binary deoxyribozyme (BiDz) probe in terms of sensitivity and selectivity. Methods: In vitro fluorescence measurements and gel shift assays were used to assess the secondary structures, sensitivity, and selectivity of the designs. Results and Discussion: All three sensors successfully detected a synthetic HPV-16 ssDNA analyte, with BiDz demonstrating the highest sensitivity and the lowest limit of detection (10 pM). DNM-I exhibited higher background fluorescence due to partial self-activation, while DNM-II showed improved background control but slightly reduced sensitivity. Both DNMs retained excellent single-nucleotide selectivity (99.9%). Conclusions: The scaffold topology was found to strongly influence sensor performance, affecting catalytic activity and background fluorescence. The introduction of displaced strand-binding elements resulted in poorer performance compared to multiple-binding armed DNMs. Although both DNMs retained high selectivity, further optimization is required to achieve efficient dsDNA recognition at physiological temperatures.

Objective: Lactoferrin is a multifunctional glycoprotein with beneficial properties, including immunomodulatory, anti-inflammatory, antibacterial, antiviral and antioxidant properties. hLF contained in breast milk provides natural protection of newborns from infections. Testing the quality of breast milk and colostrum is an important task that will enable the adjustment of the baby’s nutrition. A large number of methods for determining hLF are known, most often instrumental and immune tests are used. However, simple and rapid analysis methods are needed. Methods: 32 colostrum samples and 10 milk were obtained from healthy women. Lactoferrin content was tested by fluorescence polarization immunoassay (FPIA) and ELISA using monoclonal and polyclonal antibodies. Results and Discussion: Previously, we demonstrated the possibility of determining hLF by the FPIA using fluorescently labeled recombinant camel nanobodies, which took no more than 10 min. In this work, we demonstrate the possibility of determining hLF in colostrum samples, which differs in composition from milk. A one-step enzyme immunoassay using polyclonal and monoclonal antibodies has been developed, which allows for faster and minimal-stage determination of hLF in milk and colostrum. The ELISA detection limit was 1.25 ng/mL. 32 colostrum samples and 10 milk samples were tested using FPIA and ELISA. Conclusions: It has been shown that both methods correctly determine hLF in milk and colostrum. It is important to note that the developed ELISA can be extended to other human physiological fluids, such as tear fluid or urine.

Objective: Single embryo transfer is a strategy used in assisted reproductive technologies that can reduce the incidence of multiple pregnancies. In this approach, the importance of methods for assessing embryo quality increases substantially, as it is necessary to select the embryo with the highest implantation potential from among several embryos. One way to improve embryo quality assessment is through non-invasive methods of diagnosing oocyte competence. The aim of the study was to evaluate the potential use of the expression levels of long non-coding RNAs (lncRNAs) NEAT1, MALAT1, ANXA2P2, MEG3, IL6STP1, VIM-AS1, HSAT2/3, LTR12c-375, HERVS71-int9, MER52c-175 as predictors of oocyte quality. Methods: Nine donors and 19 patients with various IVF outcomes participated in the study. The groups of donors and patients did not differ significantly in age, body mass index, antimüllerian hormone level, duration of stimulation, or daily dose of hormones. RNA was isolated from the cumulus cells and used for cDNA synthesis and subsequent amplification by qPCR. The data were analyzed using statistical methods to determine the correlation between the level of the lncRNAs expression and IVF procedure outcome. Results and discussion: The expression levels of the lncRNAs were analyzed in relation to the IVF outcome and the embryologic status of patients and donors (total number, number of mature oocytes, and number of day 5 blastocysts). Decreased expression of NEAT1 and increased expression of VIM-AS1 were the predictors most strongly associated with successful IVF outcomes, i.e., birth (correlation coefficient r (–0.7) and 0.6 respectively). Logistic regression parameters were determined for NEAT1 to describe the relationship between IVF outcomes and non-coding RNA transcription levels (b0 = 3.852; b1 = –1.418). There was no correlation between the NEAT1 transcription level and the number of oocytes or blastocysts. This suggests that NEAT1-dependent factors that reduce oocyte competence play a role in post-implantation stages. VIM-AS1 expression showed a weak correlation with the total number of oocytes (r = 0.3). Transcript 2 isoform of the VIM-AS1 correlated also with the number of mature oocytes and day 5 blastocysts. Conclusions: NEAT1 and VIM-AS1 non-coding RNAs of cumulus cells are involved in follicle maturation processes and can be considered as potential biomarkers for assessing oocyte quality and predicting the outcome of IVF procedure.

Objective: A complex approach using test organisms from different taxonomic groups (the unicellular green alga Chlorella vulgaris and the onion Allium cepa) was employed to assess the toxicity, cytotoxicity, and genotoxicity of polycarboxylic acids containing cycloaliphatic fragments. Methods: Onion (Allium cepa L.) of the Stuttgarter Riesen variety was used as a test object. Samples were analyzed using a Mikromed microscope (Russia) at 400× magnification. Differences were considered statistically significant at p < 0.05. The unicellular green alga Chlorella vulgaris (Beijer), strain LARG-1, obtained from the Vavilov Institute of General Genetics of the Russian Academy of Sciences, was used to investigate genotoxic effects. The culture was incubated in a Flora-1 photobioreactor (Russia) for 10 days at 25°C. Visible mutations were determined using an Altami stereomicroscope (Russia) at magnifications ranging from 15× to 40×. Results and Discussion: Polycarboxylic acids I–IV were evaluated at concentrations of 1.0%, 0.1%, 0.01%, and 0.001%. Compounds I and II at a concentration of 1.0% significantly inhibited the cell division rate, almost completely stopping it. Compounds I–IV at a concentration of 0.1% inhibited the cell division rate in the onion root meristem, reducing the mitotic index to 8.33 ± 1.12, 8.11 ± 0.90, 5.33 ± 0.70, and 8.33 ± 0.28%, respectively, compared to 11.05 ± 0.73% in the control. In contrast, compounds I–IV at concentrations of 0.01 and 0.001% did not cause a significant decrease in the mitotic index or observable damage to the root meristem. Analysis of the mitotic phase indices showed that compounds I–IV at a concentration of 1.0% exhibited antiproliferative activity. The tested polycarboxylic acids demonstrated mutagenic effects in the bioassay with Chlorella vulgaris. Conclusions: The analysis of the genotoxic activity polycarboxylic acids I–IV in various concentrations carried out using a system of toxicogenetic methods, showed that at high concentrations these compounds can reduce the mitotic index and phase indexes. Although no chromosomal aberrations were observed, the compounds demonstrated significant cytotoxicity and mutagenicity, as evidenced by the inhibition of cell division in Allium cepa and the increased mutation frequency in Chlorella vulgaris. The polycarboxylic acids I–IV with antiproliferative activity can be used as promising scaffolds. These drugs can be recommended for further research. The approach described here in may be applied for obtaining rapid, cost-efficient and useful supplementary data on different types of toxicity in vitro for promising scaffolds as well as for drugs under development.

Objective: Quantitative, rapid and high-throughput analysis of IgG and IgA immunoglobulins is necessary to determine the content of these proteins and their associated molecules in the patient’s physiological fluids. The analysis of these proteins is necessary in the diagnosis of specific antibody deficiency as an auxiliary test for the detection of general variable immunodeficiency, as well as for risk stratification of patients with low IgA levels. IgG content determination can help in prescribing revaccination to patients and supporting their treatment strategy, can be used to monitor the patient's humoral immune system, as well as in the development and subsequent production of most therapeutic antibodies in biopharmaceuticals. Methods: Miniature recombinant single-domain antibodies (nanobodies) have a number of advantages over classical antibodies, such as their relative simplicity of operation, high stability over a wide range of temperature and pH values, the ability to recognize highly specific conformational epitopes of the target protein, as well as the possibility of using them as probes for detecting larger target antigen proteins in the fluorescence polarization method. Results and Discussion: Fluorescently labeled FITC-anti-IgG and FITC-anti-IgA nanobodies to human IgG and IgA were obtained and characterized. The K_D values of the FITC-anti-IgG*IgG and FITC-anti-IgA*IgA complexes were determined; they confirmed the high affinity of the immunoreagents. The possibility of specifically determining IgG and IgA levels in human serum in the range of 35– 120 μg/mL (for IgA) and 75–260 μg/mL (for IgG) was demonstrated. Eighteen human sera were tested for IgG and IgA levels, and the content of antibodies in the samples was confirmed using commercial enzyme immunoassay kits. FITC-anti-IgG and FITC-anti-IgA did not interact with other human proteins: albumin, plasminogen, fibrinogen, lactoferrin, and transferrin. Testing of human and animal sera by FITC-anti-IgG and FITC-anti-IgA demonstrated specific binding to human and monkey antisera, but not to animal sera: bovine, canine, feline, rabbit, and sheep. Conclusions: Thus, the FPIA method can be used for the rapid and specific determination of human IgG and IgA.

Objective: This article focuses on the design, synthesis, and characterization of C-19 acylamino-functionalized isosteviol derivatives, and their evaluation for LSD1 inhibitory activity as well as molecular docking studies. Methods: The novel C-19 acylamino-functionalized isosteviol derivatives were designed based on the principle of drug combination, and were subsequently synthesized via acetylation with acyl chlorides and condensation with amines. The LSD1 inhibitory activity of the synthesized compounds was evaluated using the LSD1 small molecule inhibitor screening platform. Molecular docking studies were performed using MOE (Version 2019). Results and Discussion: The synthesized C-19 acylamino-functionalized isosteviol derivatives were characterized by IR, NMR, and HR-MS spectra. The results of the anti-LSD1 activity assays showed that compounds with cyclic substituents generally exhibited excellent inhibitory activity. In particular, compound IIIl exhibited the best LSD1 inhibitory effect, with an IC₅₀ value of 8.523 ± 0.882 μM. Further molecular docking studies revealed that the 16-carbonyl group of compound IIIl formed a hydrogen bond with the Ser289 residue, and its aromatic ring formed a π-H interaction with the Met332 residue. Additionally, the top-ranked docking pose of compound IIIl showed a strong binding affinity to the LSD1 protein, with a docking score of –5.273.Conclusions: This study lays the groundwork for the development and structural modification of new isosteviol-based drugs.