We present a new issue of the "Leadership Opinion" series, dedicated to strategic projects in the fields of biotechnology and immunoprophylaxis.

Polar steroid compounds from starfish and triterpene glycosides from sea cucumbers represent unique classes of biologically active secondary metabolites characterized by considerable structural diversity and a broad range of biological effects. A metabolomics approach based on HPLC-MS/MS was applied to investigate complex mixtures of these natural compounds. Chromatographic separation was performed using a LicArt 62 HPLC coupled to a Bruker Impact II Q-TOF quadrupole time-of-flight mass spectrometer. More than 350 compounds were detected in the Lethasterias fusca starfish extract and more than 550 compounds were detected in the Eupentacta fraudatrix sea cucumber extract. 

In the development of any analytical method, a key step is the assessment of its robustness, i.e., its ability to remain unaffected by minor variations in different parameters. Given sufficient time, a method developer can thoroughly investigate each factor affecting chromatographic separation, including mobile phase composition, column temperature, injection volume, and other parameters. However, one of the most significant factors determining retention time, peak shape, and resolution is the chromatographic system itself. When developing a new method, a researcher can readily test it using instruments from different manufacturers. However, when transferring an already approved analytical method to a new instrument under time constraints, there is a risk that the chromatographic system may fail to meet the established system suitability criteria.

Introduction. The high prevalence of pain syndromes among the global population highlights the need for searching for highly potent analgesic agents with a favorable safety profile. 3-Hydroxy-3-pyrrolin-2-one derivatives exhibit diverse pharmacological activities and represent promising compounds for the development of novel analgesic drug candidates with pronounced efficacy.

Aim. To study the degree of analgesic potential of previously unknown 5-aryl-4-aroyl-1-isobutyl-3-hydroxy-3-pyrrolin-2-ones in a hot plate test.

Materials and methods. Newly synthesized 3-hydroxy-3-pyrrolin-2-ones containing an isobutyl substituent at position 1 (I–VI) were the objects of the study. The analgesic efficacy of 6 analog compounds was assessed in vivo using a behavioral nociception model.

Results and discussion. Pharmacological screening demonstrated that the new compounds previously synthesized by the authors exhibited analgesic potential following oral administration in thermal nociception tests. Compound VI demonstrated the most pronounced analgesic activity. Structure–activity relationship (SAR) analysis revealed distinct patterns correlating molecular features with analgesic efficacy. These findings indicate that the newly synthesized 3-hydroxy-3-pyrrolin-2-ones bearing an isobutyl substituent at position 1 exhibit significant analgesic potential (compounds I, III, V, VI).

Conclusion. The obtained 5-aryl-4-aroyl-1-isobutyl-3-hydroxy-3-pyrrolin-2-ones (a series of 6 analog compounds) were tested on a model of behavioral nociception, the relationship between the chemical structure of the substances and their activity was studied, and the main candidate with significant analgesic potential was identified for further study (laboratory sample with code VI).

Introduction. Fungi represent an evolutionarily distinct group of heterotrophic organisms that occupy a unique position in the system of wildlife. Currently, they are considered by modern medicine as promising sources of new biologically active compounds. Despite a millennia-long history of use in traditional medicine and an increasing number of studies on the biological activity of various groups of fungal substances, the incorporation of this raw material into pharmaceutical practice remains limited. The key problem lies in the difficulty of standardization due to the absence of regulatory documentation. Unlike plants, the number of pharmacopoeial monographs on fungal raw materials (fruit bodies, mycelium, extracts) is very low, and unified approaches to quality assessment are lacking. Thus, the development and harmonization of standards for fungal raw materials will enable the realization of their therapeutic potential.

Text. This article addresses the issues of standardization of fungal raw materials. The main areas of application of macromycetes are described. The principal species of higher fungi used in medicine are named, and the prospects for using other potential candidates are discussed. The types of confirmed biological activity for specific fungal species, including antitumor, immunomodulatory, and antioxidant activities, are described. The increasing number of studies on higher fungi as sources of biologically active compounds is demonstrated. The pharmacopoeias of Russia, China, Japan, India, the USA, Brazil, and other countries were analyzed for the presence of monographs on mushrooms. The materials for this review include various pharmacopoeial editions available in open access in English and Russian. The species of pharmacopoeial fungi are identified, along with similarities and differences in standardization approaches. Quality parameters used for standardization in different pharmacopoeias are presented. Examples of medicinal products and biologically active food additives based on mushrooms registered in Russia are given. The problems of developing regulatory documentation, quality standards, and analytical methods for fungal raw materials are outlined.

Conclusion. It has been determined that the regulatory framework governing the quality of fungal raw materials is extremely limited. The expansion of the lists of pharmacopoeial fungal species used for the production of medicines and dietary supplements is necessary.

Introduction. Sosnowsky's hogweed (Heracleum sosnowskyi Manden.) is a dangerous invasive transformer species widespread in the European part of Russia. The inclusion of the plant in the list of hazardous invasive species from 2026 and the ongoing regional control programs necessitate the rational utilization of its biomass. A detailed study of the phytochemical profile is a key prerequisite for justifying processing prospects.

Aim. To study the phytochemical profile of H. sosnowskyi biomass, including the isolation of furanocoumarins, polysaccharides, and essential oils of pharmaceutical value, to provide a scientific basis for the rational use of this invasive species.

Materials and methods. The objects of the study were the herb and fruits of H. sosnowskyi collected in Perm Krai and Leningrad Oblast in 2024. Furanocoumarins were extracted with n-hexane (Soxhlet apparatus), purified by preparative HPLC; structures were confirmed by UV, NMR, and high-resolution mass spectrometry. Polysaccharide fractions (water-soluble polysaccharides – WSP, pectic substances – PS, hemicelluloses) were obtained by sequential extraction with water, ammonium oxalate, and alkali, then purified using the Sevag method. Yield was assessed gravimetrically, monosaccharide composition by HPTLC, structural features by IR spectroscopy, and molecular weights by size-exclusion chromatography. Essential oil from the fruits was obtained by hydrodistillation (Clevenger apparatus), and its component composition was analyzed by GC-MS.

Results and discussion. Five furanocoumarins – xanthotoxin, psoralen, angelicin, isopimpinellin, and bergapten – were isolated from the aerial part of H. sosnowskyi and identified. The total polysaccharide content in the herb was 4.1 %, with the yield of the PS fraction at 2.3 ± 0.2 % and the WSP fraction at 1.8 ± 0.1 %. The dominant monosaccharide in both fractions was arabinose. The average molecular weight of WSP was ~160 kDa, and PS was ~193 kDa. The yield of essential oil from the fruits was 1.01 ± 0.03 %; the main component was octyl acetate (61.8 %), along with hexyl butyrate (6.4 %), 2-methylhexyl butyrate (6.0 %), and hexyl isobutyrate (5.5 %).

Conclusion. H. sosnowskyi biomass is a promising multicomponent source of biologically active compounds. The proposed integrated processing approach allows the sequential isolation of furanocoumarins (5 compounds) and polysaccharides (4.1 %) from the herb, while the fruits serve as a source of essential oil with high octyl acetate content. These results provide a scientific basis for the rational use of this invasive species for pharmaceutical purposes.

Introduction. In situ forming implants (ISFIs) are liquid drug delivery systems that form a solid polymer matrix directly at the injection site in response to physiological stimuli. These systems provide prolonged localized release of active pharmaceutical ingredients (APIs), making them promising for the treatment of various diseases. A key challenge in their pharmaceutical development is the lack of reliable predictive models linking in vitro release kinetics with in vivo performance.

Text. This review provides a systematic and critical analysis of existing in vitro methods for assessing drug release from ISFIs, approaches to selecting biorelevant media, mathematical modeling, and substantiates the main directions for establishing in vitro – in vivo correlation (IVIVC). The main groups of release testing methods, such as the Dissolution test using various apparatuses, dialysis methods, and the sample-and-separate method, as well as more modern approaches including the medium replacement method and diffusion in an agarose matrix, are considered. It is shown that the choice of a biorelevant dissolution medium, considering the specifics of the microenvironment at the implantation site (synovial, gingival, interstitial, tear fluid, tumor microenvironment), is a critically important factor for obtaining prognostically valuable data. The capabilities of classical (zero-order, first-order, Higuchi, Korsmeyer – Peppas) and mechanistic mathematical models for describing the complex release kinetics, determined by the interrelated processes of diffusion, swelling, and degradation of the polymer matrix, are analyzed. Special attention is paid to the analysis of precedents for establishing IVIVC for ISFIs, from the first linear correlation to the world's first successfully developed and validated Level A correlation.

Conclusion. Establishing IVIVC for ISFIs is not fundamentally impossible but requires systematic engineering of in vitro methods with an emphasis on biomimetic adjustment of geometric parameters and medium composition. The key directions for future research are the creation of integrated test systems, the incorporation of biological factors into protocols, and the application of non-invasive monitoring techniques.

Introduction. Paclitaxel is a plant-derived antineoplastic agent with high potential in the treatment a wide range of malignant neoplasms. However, the unique properties of paclitaxel – low aqueous solubility, poor oral bioavailability, and high toxicity – create considerable challenges in the development of stable formulations, substantially limiting its clinical application. Therefore, there is an active ongoing search for novel technologies capable of enhancing the therapeutic efficacy of paclitaxel while reducing the impact of toxic effects on the human body.

Text. Paclitaxel is one of the most widely used drugs for the treatment in modern oncology. Its antitumor mechanism of action is based on its ability to act on tubulin, stimulating the assembly of microtubules, stabilizing them, and thus preventing their dissociation, which leads to mitotic arrest and subsequent cell death. Paclitaxel possesses a complex structure comprising various functional groups, including ester linkages, a taxane ring, and aromatic moieties. These molecular elements determine paclitaxel's anticancer activity as well as its physicochemical properties. Paclitaxel belongs to Class IV according to the Biopharmaceutics Classification System (BCS) and has low solubility and low intestinal permeability. Drug substances of this class are the most challenging for formulation development and exhibit extremely low bioavailability. The low solubility, poor bioavailability, and high toxicity of paclitaxel pose significant challenges in pharmaceutical development. Furthermore, the polymorphism of paclitaxel can also have a significant influence on stability and biopharmaceutical properties of finished dosage forms. For example, the choice of polymorphic form can be of fundamental importance for the efficacy and safety not only of antitumor drugs but also of paclitaxel coated medical devices used in the treatment of cardiovascular diseases, in particular restenosis after angioplasty. Despite the availability of formulations successfully used in clinical practice, the development of new ones continues to this day. Nowadays, researchers face the task of enhancing paclitaxel's bioavailability while reducing its toxic effects on the human body.

Conclusion. The challenges in developing paclitaxel formulations are multifaceted. New strategies for treating malignant tumors using paclitaxel are in place, and novel drug compositions and combinations are under investigation. This work reviews contemporary studies focused on the development of new dosage forms of paclitaxel.

Introduction. Low viability of probiotic microorganisms during gastrointestinal (GI) transit substantially limits the therapeutic efficacy of oral probiotic formulations. Incorporating the prebiotic inulin into the microcapsule matrix and employing different types of hard capsules may further enhance bifidobacterial protection; however, a comprehensive comparison of these factors using a single GI model has not been conducted.

Aim. To evaluate the effect of inulin incorporation into alginate-chitosan microcapsules and the type of hard capsule (gelatin / enteric-coated) on the viability of B. bifidum under simulated GI conditions.

Materials and methods. Six experimental formulations were prepared: microcapsules (alginate + chitosan) without and with inulin, placed in gelatin and enteric-coated capsules, as well as control groups (bifidumbacterin powder in gelatin and enteric-coated capsules). GI simulation was performed according to the State Pharmacopoeia of the Russian Federation (XV edition, General Monograph 1.4.2.0014): acid stage (0.1 M HCl, pH 1.2, 2 h) followed by buffer stage (pH 6.8). Viability was assessed by Colony-Forming Unit (CFU) counting on MRS-5 agar.

Results and discussion. The highest survival rate was observed in the sample "microcapsules with inulin in an enteric-coated capsule". Microcapsules with inulin in a gelatin capsule provided comparable survival. Microcapsules without inulin demonstrated a slight reduction in cell titer (both enteric-coated and gelatin capsules). Bifidumbacterin powder in a gelatin capsule was characterized by a reduction of viable CFU by 2 orders of magnitude, while in an enteric-coated capsule without microencapsulation, survival decreased by 1 lg CFU/capsule.

Conclusion. Alginate-chitosan microencapsulation is the main factor protecting B. bifidum. Incorporation of inulin into the matrix additionally increases survival. The combination of microencapsulation with inulin and an enteric-coated capsule provides maximum protection; however, no statistically significant advantage of the enteric-coated capsule over the gelatin capsule was detected in the context of microencapsulation with inulin (p > 0.05).

Introduction. Aesculus hippocastanum L. seeds are a promising source of triterpene saponins – escin (β-escin) possessing venotonic, angioprotective, and anti-inflammatory activities. The production of β-escin substance in Russia is currently absent, which determines the relevance of developing a reproducible technology for obtaining purified horse chestnut seed extract with a high escin content.

Aim. To determine the optimal extraction parameters of triterpene saponins from Aesculus hippocastanum L. seeds using multivariate optimization, to develop an extract purification procedure, and to establish the Normal Operating Range of process parameters.

Materials and methods. Extraction process optimization was performed according to a Box – Behnken design. The effects of ethanol concentration, temperature, solid-to-solvent ratio, and extraction time on the yield of total triterpene saponins expressed as β-escin were investigated. The chemical composition of the extract was determined by HPLC-MS with ESI+/ESI− detection. The quantitative content of saponins in the extract was determined by spectrophotometry; β-escin content in the purified extract was determined by HPLC.

Results and discussion. A quadratic regression model of the extraction process was developed. The solid-to-solvent ratio and ethanol concentration were identified as the primary factors affecting saponin yield, while temperature within the studied range was found to be statistically non-significant. The optimal extraction conditions were: ethanol concentration – 49 %, temperature – 45 °C, solid-to-solvent ratio – 1 : 12, extraction time – 90 min; the predicted yield was 75.6 %. In accordance with ICH Q8(R2) principles, the Design Space (DS) (Y ≥ 65 %) and the Normal Operating Range (NOR) were established. Seven structural groups of escins were identified in the purified extract by HPLC-MS.

Conclusion. The Design Space and Normal Operating Range for the extraction process of triterpene saponins from Aesculus hippocastanum L. seeds were established within the ICH Q8(R2) / QbD framework, ensuring the production of purified extract with reproducible quality attributes.

Introduction. The development of oral dosage forms with controlled release remains an important area of modern pharmaceutical technology. Such systems can reduce fluctuations in plasma drug concentration, decrease dosing frequency, and improve the convenience of long-term therapy. Among modified-release platforms, osmotic drug delivery systems are of particular interest because the release of an active pharmaceutical ingredient is governed not only by its physicochemical properties but also by predefined engineering parameters of the dosage form. These parameters include the core composition, osmotic gradient, semipermeable membrane properties, the presence and size of the delivery orifice, and the characteristics of the push layer, etc.

Text. This review aims to analyze OROS technology as an engineering platform for controlled drug release, with consideration of the main structural types of osmotic systems, their mechanisms of action, critical formulation factors, and technological limitations.

Conclusion. The analysis of literature and patent data shows that OROS remains a relevant approach to the development of modified-release dosage forms owing to its ability to provide programmable and reproducible drug release. The elementary osmotic pump, push-pull osmotic pump and controlled-porosity osmotic pump are of the greatest practical importance, as these systems address different technological tasks: delivery of soluble drugs, release of poorly soluble active pharmaceutical ingredients, and formation of a microporous release pathway without a separate laser-drilling step. At the same time, OROS systems require careful pharmaceutical design, control of critical quality attributes, and specialized manufacturing equipment. These factors limit their widespread implementation but maintain strong interest in this technology for the development of modern prolonged-release oral drug products.

Introduction. Chronic, burn, postoperative and infected wounds require dressings that not only maintain a moist environment, but also help control bacterial contamination, inflammation, exudation and tissue regeneration. Gel and hydrogel dressings based on animal-derived biopolymers are of interest for pharmaceutical industry because their properties can be adjusted through the choice of raw material, crosslinking method, matrix composition and various modifications.

Text. This review discusses six biopolymer platforms: collagen, gelatin, chitosan, hyaluronic acid, keratin and silk fibroin. The focus is placed not only on their antibacterial and reparative properties, but also on the technological parameters that determine their practical applicability: molecular weight, degree of modification, gelation time, swelling, mechanical strength, adhesion to wet tissue, release profile, biodegradation, sterility, stability and cytocompatibility. Crosslinking methods, approaches to raw material standardization, choice of sterilization method, quality control, the level of preclinical and clinical evidence, and regulatory aspects of medical devices and combination products are also considered.

Conclusion. The potential of a hydrogel dressing is determined not only by the biological activity of the material, but also by the reproducibility of its critical quality attributes. The most relevant platforms for further development are those in which composition, manufacturing technology, sterilization, release of active components, safety and intended clinical use are linked at the design stage.

Introduction. Transdermal Therapeutic Systems (TTS), particularly transdermal patches, represent one of the most promising non-invasive routes for delivering drugs into the systemic circulation. A key prerequisite for realizing the therapeutic potential of transdermal patches is the maintenance of stable and uniform contact between the system and the skin surface throughout the intended period of use, achieved through optimal adhesive characteristics. The adhesion of TTS is a critical quality attribute (CQA) that determines drug bioavailability and patient safety, and it requires objective assessment that takes into account skin biomechanics and real-use conditions.

Aim. A comparative analysis of adhesive properties of transdermal patches with the phytoestrogen trans-resveratrol using the probe tack test.

Materials and methods. The objects of the study were matrix-type patch samples obtained at the St. Petersburg State Chemical and Pharmaceutical University (SPCPU) of the Ministry of Health of the Russian Federation, containing trans-resveratrol. Adhesion was evaluated by probe tack test using the texture analyzer TA.XTplus with Exponent Connect software. The study was conducted at the Institute of Pharmacy, Kazan State Medical University (KSMU) of the Ministry of Health of the Russian Federation. Transdermal patches Nicorette® and Evra® (LTS Lohmann Therapy-Systems AG, Germany) were used as comparator products (positive controls). Two types of substrates were used to model human skin: a biological substrate (isolated pig skin) and a synthetic substrate (silicone skin). The test results were processed using standard statistical methods in accordance with the requirements of the State Pharmacopoeia of the Russian Federation.

Results and discussion. The patch sample based on a copolymer of methacrylic acid and ethyl acrylate as a polymer carrier acting as an adhesive demonstrated the highest tack, comparable to that of the reference products. The incorporation of polyvinylpyrrolidone K-17 to enhance the release of trans-resveratrol from the matrix resulted in a partial loss of patch adhesion and the formation of an unstable system that was highly sensitive to environmental conditions.

Conclusion. Based on the evaluation of peak force required to break the bond and work of adhesion, it was established that the use of a copolymer of methacrylic acid and ethyl acrylate for the development of matrix-type transdermal patches with trans-resveratrol represents a promising approach. This strategy enables an optimal combination of adhesive properties, comparable to those of reference products, while maintaining the active substance release performance from the TTS as reported in a previous study.

Introduction. Methacrylic acid copolymers are synthetic anionic copolymers that are used as the main component of enteric-soluble coatings that provide controlled drug release in the small intestine. The molecular weight of these copolymers is a critical parameter affecting their solubility in the gastrointestinal tract, therefore it is among the normalized parameters that determine the quality of copolymers.

Aim. Development of a method for determining the weight average molecular weight of methacrylic acid copolymers by diffusion-ordered NMR spectroscopy, which does not involve the stage of converting the molecular weight of derivatives into the molecular weights of the initial methacrylic acid copolymers.

Materials and methods. A solution of the sodium salt of polymethacrylic acid, copolymers of methacrylic acid from various manufacturers, as well as copolymers of methacrylic acid isolated from pharmaceutical film coatings were used as objects of research. The DOSY experiments were performed on an Agilent DD2 600 NMR spectrometer.

Results and discussion. Using a set of comparison samples, a calibration graph of the dependence of the logarithm of the weight average molecular weight of the comparison samples on the logarithm of their adjusted average value of the self-diffusion coefficient was constructed and a correlation equation with a coefficient of determination R² = 0.9992 was derived. The values of the molecular weights of the tested samples calculated using this correlation equation agreed within the confidence intervals with the values measured by the coating manufacturers using the GPC method. The relative measurement error of the DOSY method (5.6 %) does not exceed the relative error of the determination of molecular weights by the GPC method (5–10 %).

Conclusion. In the course of the study, a method was developed for determining the weight average molecular weight of methacrylic acid copolymers by diffusion-ordered NMR spectroscopy. When implementing this technique, the possibility of formation of methacrylic acid copolymers conglomerates due to their derivatization into the corresponding sodium salts is excluded; recalculation of the weight average molecular weight of the derivatized methacrylic acid copolymers into the weight average molecular weight of the initial methacrylic acid copolymers is excluded; different mobility of individual macromolecule segments is taken into account; the effect of the viscosity of the analyzed solutions on the measured value of the self-diffusion coefficient has been reduced. The relative error in measuring the weight average molecular weight of methacrylic acid copolymers using the developed technique does not exceed the relative error in measuring the molecular weights of substances by the GPC method.

Introduction. Acetochlor is one of the most common herbicides capable of accumulating in medicinal plant raw materials and posing a health risk. A method for acetochlor determination was developed using fluorescence polarization immunoassay (FPIA), a homogeneous and specific technique employed for analysis of low-molecular-weight toxicants.

Aim. To develop and validate a fluorescence polarization immunoassay for quantitative analysis of the herbicide acetochlor in medicinal plant material, exemplified by corn silk (Zea mays L.).

Materials and methods. For the development of the method, polyclonal rabbit antibodies against acetochlor and a tracer consisting of an acetochlor derivative conjugated with ethylenediamine fluorescein (AMPA-EDF) were used. The analysis was performed by fluorescence polarization using a portable Sentry-200 fluorimeter (Ellie LLC, USA). Data processing was carried out with SigmaPlot 11 software package (Systat Software Inc., USA). The study was performed on samples of corn silk medicinal plant material purchased from a local pharmacy and an online store. Method validation included the following parameters: limit of detection, limit of quantification, analytical range, precision, accuracy, and specificity.

Results and discussion. A method for the quantitative determination of the pesticide acetochlor in the herbal medicinal raw material corn silk (Zea mays L.) has been developed, optimized, and validated using fluorescence polarization immunoassay (FPIA). The determination range was from 0.024 to 1.56 mg/kg. The specificity, precision, and accuracy of the developed method were confirmed.

Conclusion. The fluorescence polarization immunoassay (FPIA) method was applied for the quantitative determination of the pesticide acetochlor in herbal medicinal raw material, using corn silk as an example.

Introduction. Phytotherapy represents an integrative method for disease prevention and treatment using complexes of biologically active substances (BAS) of plant origin. Aesculus hippocastanum L. is recognized as a valuable source of plant material: its seeds are used in evidence-based medicine for venous insufficiency treatment, while flowers are utilized in traditional practice due to their pronounced venotonic, anti-inflammatory, antioxidant, and hypolipidemic properties. This establishes Aesculus hippocastanum L. flowers as promising medicinal plant raw material (MPRM) with substantial therapeutic potential. Liquid dosage forms (LDF) based on MPRM possess multiple advantages ensuring their demand in pharmaceutical practice and among consumers. Accordingly, developing new domestic phytopreparations as infusions, tinctures, and liquid extracts from these flowers with potential venotonic, capillary-protective, and antioxidant effects constitutes a relevant direction in pharmacy.

Aim. To obtain and standardize experimental laboratory samples of aqueous and aqueous-alcoholic extracts from Aesculus hippocastanum L. flowers for the subsequent substantiation of the optimal liquid dosage form.

Materials and methods. A comparative study of aqueous and aqueous-alcoholic extracts from Aesculus hippocastanum L. harvested in the Voronezh Region during the flowering period of 2024 was conducted for infusions (1 : 10), tinctures (1 : 5) prepared by maceration, fractional maceration, and percolation methods, and liquid extract (1 : 1) according to the State Pharmacopoeia of the Russian Federation, XV edition (GPM.1.4.1.0019 «Tinctures», GPM.1.4.1.0021 «Extracts», GPM.1.4.1.0018 «Infusions and decoctions»). For the obtained LDF laboratory samples, identification and quantitative determination methods for flavonoids, saponins, and antioxidants were developed with yield assessment relative to raw material content, alongside standardization by «Description», «Dry residue», «Ethyl alcohol», and «Heavy metals» parameters per relevant GPM requirements.

Results and discussion. Experimental aqueous and hydroalcoholic LDF samples (infusion, tinctures, liquid extract) based on Aesculus hippocastanum L. flowers were obtained and standardized according to regulatory documentation requirements. Methods for quantitative determination of target BAS groups – flavonoids, saponins, antioxidant activity, and total antioxidants expressed as ascorbic acid – were developed. The liquid extract (1 : 1) showed the highest content of antioxidants, flavonoids, and saponins, recommending it as the optimal form among those studied.

Conclusion. Aesculus hippocastanum L. flowers represent promising MPRM for developing effective and safe domestic phytopreparations. Due to high BAS concentration, liquid extract is recommended as an oral dosage form and component of topical semisolid dosage forms, aligning with current phlebotonic drug market trends.

Introduction. Bacopa monnieri (Brahmi) is an Ayurvedic medicinal plant with pronounced nootropic and neuroprotective properties. The therapeutic effect of preparations based on it is mainly associated with the content of triterpene saponins, but other components, including phenolic compounds, contribute to the overall effect. Despite the long history of medicinal use, there are few studies of the diversity of biologically active substances (BASs) of B. monnieri.

Aim. Analysis of the component composition of biologically active substances of Bacopa monnieri herb (aerial parts) extracted with 80 % ethyl alcohol using ultra-performance liquid chromatography with mass spectrometric detection (UPLC-MS).

Materials and methods. The study subject was B. monnieri herb (Hassan, India). A 1 : 10 extract was obtained by threefold extraction with 80 % ethyl alcohol. The qualitative composition of the obtained extract was studied by UPLC-MS on a Waters Acquity chromatograph with a diode-array UV detector and a TQD MS detector (Waters Corporation, USA) under the following conditions: Acquity UPLC BEH C18 column (0.21 × 15.0 cm, 1.7 μm), gradient elution mode (solvent A – water-acetonitrile mixture (95 : 5) with 0.1 % formic acid, solvent B – acetonitrile with 0.1 % formic acid), column temperature 35 °C, flow rate 0.25 mL/min, UV detection at 220–500 nm, mass spectrometry in the positive and negative ion registration modes.

Results and discussion. In 80 % ethanol extract of B. monnieri herb, 14 phenolic compounds were detected, of which 7 were identified (hydroxycinnamic acids and their glycosides: caffeic acid 4-O-glucuronide, monnieraside III, plantainoside B, 1-(3,4-dihydroxyphenethyl)-2-feruloyl-D-glucoside, flavonoids: quercetin-3-glucuronide, luteolin-7-glucuronide, apigenin-7- glucuronide). Targeted search for saponins at 455 selected ion mass detected 12 compounds and identified 6 of them: bacoside A3, bacopasides II and VII, bacomosaponins A and B, bacopasaponin C.

Conclusion. The composition of an 80 % ethanol extract of B. monnieri was studied using UPLC-MS. Luteolin-7-glucuronide, quercetin-7-glucuronide, caffeic acid 4-O-glucuronide and 1-(3,4-dihydroxyphenethyl)-2-feruloyl-D-glucoside were identified for the first time in the raw material of this species.

Introduction. Amino acids (AAs) are key components of living organisms. Medicinal plants are valuable sources of AAs, which enhance the bioavailability of other metabolites and potentiate their pharmacological effects. However, the amino acid composition of many plants, including Oxytropis pilosa (L.) DC., remains unexplored.

Aim. To establish the qualitative and quantitative composition of free and bound amino acids in O. pilosa raw material growing in the Saratov region.

Materials and methods. O. pilosa raw material collected during the period of mass flowering in the Saratov region was used as the object of the study. Qualitative analysis of free AAs was performed by thin-layer chromatography (TLC) using an n-butanol – acetone – glacial acetic acid – water system (35 : 35 : 10 : 20) with 2 % ninhydrin in alcohol as the detection reagent. Quantitative determination of the total free AAs was carried out by spectrophotometry using a Shimadzu UV-1800 spectrophotometer (Japan) at an analytical wavelength of 568 nm. Qualitative and quantitative analysis of free and bound AAs was performed by HPLC on a Shimadzu LC-20 Prominence chromatograph equipped with an SPD-M20A detector (Japan), an SIL-20A autosampler, and LabSolutions data acquisition and processing software. Reference standards of AAs were used for compound identification. AA content was expressed in mg/g of dry raw material. The obtained data were statistically processed using MS Excel 2017.

Results and discussion. The composition of free AAs in aqueous extracts from the herb, flowers, and leaves of O. pilosa was determined by TLC. Six AAs (aspartic acid, proline, serine, threonine, valine, and isoleucine) were identified in all studied samples. The highest total free AA content was found in the herb of O. pilosa (30.70 ± 0.39 mg/g). HPLC analysis of AAs in the extract from the herb after preliminary hydrolysis revealed 15 AAs, among which the predominant ones were alanine (66.056 ± 0.584 mg/g), 4-hydroxyproline (25.912 ± 0.278 mg/g), proline (17.834 ± 0.168 mg/g), and histidine (13.030 ± 0.086 mg/g).

Conclusion. The qualitative and quantitative composition of free and bound AAs in O. pilosa raw material has been established for the first time. The total amino acid content was 165.659 ± 0.806 mg/g, with essential AAs accounting for 16.958 ± 0.139 mg/g.

Introduction. Granik^® (the active substance is an indolinone derivative GRS) is a new domestic nitric oxide-independent stimulator of soluble guanylate cyclase (sGC). Stimulation of sGC represents a promising approach to the therapy of cardiovascular diseases caused by thrombosis, particularly in cases where traditional antiplatelet and anticoagulant agents are contraindicated and pose a high risk of bleeding. Granik^® has been in Phase I clinical trials, and this publication is devoted to the evaluation of its safety and pharmacokinetics in healthy volunteers.

Aim. The aim of the study was to evaluate the tolerability, safety and pharmacokinetics of Granik^® 20 mg capsules in healthy volunteers after administration under fasting conditions at doses of 20, 40, and 60 mg over a 7-day period.

Materials and methods. A double-blind, placebo-controlled study involved 48 healthy male volunteers who received Granik^® at doses of 20, 40, and 60 mg or placebo for 7 days. Tolerability was assessed as good, satisfactory, or unsatisfactory based on subjective data from the volunteers. Safety assessment was based on the analysis of adverse events, laboratory values, ECG data, and vital signs monitoring. GRS plasma concentrations were determined by high-performance liquid chromatography with tandem mass spectrometric detection. Statistical analysis was performed using the R v.3.5 program (Revolution Analytics, USA) and parametric and nonparametric statistics depending on the nature of the data distribution.

Results and discussion. Granik^® at doses of 20, 40 and 60 mg demonstrated good tolerability and safety after 7-day administration to healthy volunteers (16 participants in 3 cohorts). Pharmacokinetic parameters varied significantly depending on the dose. The maximum plasma concentration (C_max) of the GRS compound was reached in 0.5–1.5 h. When Granik^® was administered at a dose of 60 mg, the GRS substance remained in plasma for a long time: the half-elimination period (t_1/2) was 24.61 ± 10.40 h. When Granik^® was administered at doses of 20 and 40 mg, elimination from plasma was accelerated: the half-elimination period decreased to 4.34 ± 3.61 h and 3.69 ± 5.09 h, respectively. The accumulation coefficient of the GRS substance did not statistically significantly differ when Granik^® was administered in the dose range of 20–60 mg per day.

Conclusion. Granik^® demonstrated good tolerability and a favorable safety profile when administered as a course. These results justify the transition to the next phase of clinical trials of Granik^®.

Introduction. This study assessed the nephroprotective potential of malonic acid derivatives (MADs) in acute and subacute ischemic renal injury. A model of prerenal failure was simulated using 60-minute warm renal ischemia.

Aim. To study the nephroprotective effect of MADs in acute and subacute ischemic renal failure.

Materials and methods. The experiment was conducted using a model of warm renal ischemia induced by 60-minute occlusion of the right renal artery. During a month-long monotherapy course, the effects of 4-[(3-ethoxy-3-oxopropanoyl) amino]benzoic acid (etmaben, 60 mg/kg) and sodium 4,4'-(propanediamido)dibenzoate (maloben, 60 mg/kg) were compared with the reference drug, ethylmethylhydroxypyridine succinate (Mexidol®, 50 mg/kg). To assess renal function on days 3 and 30 after ischemia, serum creatinine and urea concentrations, glomerular filtration rate (GFR), creatinine clearance, electrolyte excretion rate, and antioxidant gene expression were analyzed, and histological examination was performed.

Results and discussion. The obtained data indicate the significant efficacy of MADs in the treatment of acute ischemic kidney injury. In the group of animals receiving etmaben, a statistically significant decrease in serum creatinine and urea levels, as well as an increase in glomerular filtration rate, was observed, with no significant effect on diuresis. Maloben, in turn, had a more pronounced effect on diuresis and electrolyte excretion during the acute period. Both studied compounds demonstrated the ability to improve renal excretory function, manifested by increased creatinine elimination and a reduction in the severity of the oligoanuric phase of acute nephropathy.

Conclusion. It was found that the studied drugs activate renal excretory function, contribute to the normalization of creatinine clearance and the attenuation of the manifestations of acute oligoanuric syndrome, stabilizing the condition of animals with acute kidney injury.

Introduction. In preclinical studies using models of nonalcoholic steatohepatitis (NASH), L-ornithine L-aspartate (LOLA) at 1.5 g/kg/day, a dose equivalent to the recommended human dose of 9 g/day, attenuated not only liver injury but also neuropsychiatric and skeletal muscle complications. The possibility of using higher LOLA doses may be of interest in the development of new approaches to the pharmacotherapy of NASH.

Aim. To evaluate the effects of a twofold LOLA dose (3 g/kg/day) on liver injury and associated extrahepatic disorders in mice with experimental NASH.

Materials and methods. Two-month-old male C57BL/6 mice were randomized into three groups: 1) Intact (n = 20); 2) Control: diet- and toxin-induced NASH model (n = 25); 3) LOLA: NASH model + LOLA administered via oral gavage at 3 g/kg once daily (n = 25). After 3 months of NASH induction, blood ammonia and lactate, urinary urobilinogen and bilirubin were measured, and the Quick-Pytel functional test was performed. Animal behavior and cognitive function were assessed using the "Open field", "Light-dark box", "Spontaneous alternation in the T-maze", and "Novel object recognition" tests. Liver tissue was examined for the severity of inflammation, macrovesicular and microvesicular steatosis, necrosis, and fibrosis. Barrier function of the jejunum and colon was evaluated using the Ussing chamber.

Results and discussion. Course administration of LOLA prevented the development of hyperammonemia (p < 0.05) and bilirubinuria (p < 0.01), ameliorated anxiety- and/or depression-like behavior (p < 0.05, p < 0.01), and improved short-term recognition memory (p < 0.05) in mice. In the liver tissue of LOLA-treated mice, a significant reduction in necrosis severity was observed (p < 0.05). LOLA administration also prevented the increase in jejunal epithelial permeability (p < 0.05), but had no significant effect on the barrier function of the colon.

Conclusion. The obtained data suggest potentially comparable efficacy of standard and twofold daily doses of LOLA in mice. The feasibility and safety of high-dose LOLA regimens require further evaluation in direct comparative studies.

Introduction. Virotherapy is one of the most promising and rapidly developing approaches in antitumor therapy. An antitumor drug based on the recombinant vaccinia virus strain VV-GMCSF-Lact, developed by a team of researchers of the Institute of Chemical Biology and Fundamental Medicine SB RAS and the State Research Center of Virology and Biotechnology "Vector", has demonstrated high antitumor efficacy against human and animal tumors. For further pharmaceutical development of VV-GMCSF-Lact evaluation of its systemic toxicity and pharmacological safety in preclinical studies is required.

Aim. Evaluation of the general toxicity and pharmacological safety of an antitumor drug based on the recombinant strain of vaccinia virus VV-GMCSF-Lact.

Materials and methods. A non-toxic dose level study in females of ICR mice, Wistar rats, and California rabbits after intravenous (iv) administration at the maximum possible doses for each animal species was conducted. Acute toxicity of the drug was evaluated in ICR mice and SD rats with a single iv or subcutaneous (sc) administration of the drug at two doses: 1 × 10⁷ PFU (human equipotent therapeutic dose) and the maximal dose for each animal species. Subchronic toxicity was studied in SD rats and New Zealand White rabbits with multiple sc administration (4 injections at 1-week intervals) of the drug at doses of 1 × 10⁷ PFU and 5 × 10⁷ PFU. The pharmacological safety of the drug was assessed in SD rats as part of subchronic toxicity studies.

Results and discussion. When determining the non-toxic dose level of the oncolytic virus VV-GMCSF-Lact with proven antitumor activity, doses exceeding the single human therapeutic dose (1 × 10⁷ PFU) by 7.5 times (mice), 30 times (rats), and 150 times (rabbits) were achieved. The observed clinical signs indicated that the achieved doses were close to the maximum tolerated. Acute toxicity studies showed that subcutaneous administration of the drug was safe at doses equivalent to the human therapeutic dose, as well as at 5-fold (mice) and 20-fold (rats) the human therapeutic dose. Intravenous administration at the indicated doses could induce a response from the organs of the immune system. Subchronic toxicity study showed that the drug was safe at doses of 1 × 10⁷ PFU and 5 × 10⁷ PFU and did not exert toxic effects on motor activity, the cardiovascular system, or the respiratory system.

Conclusion. Preclinical studies of the general toxicity and pharmacological safety of a drug with proven antitumor activity, developed on the basis of the recombinant VV-GMCSF-Lact strain of the vaccinia virus, showed that the drug is safe at the doses used and can be recommended for clinical trials.

Introduction. The pharmaceutical industry, while ensuring public health, simultaneously creates significant environmental risks. The article discusses modern approaches to assessing the impact of drugs on the environment in various regulatory systems. This impact is manifested through emissions of pollutants into the air, wastewater discharge, solid waste generation and consumption of significant volumes of natural resources as raw materials for chemical and biological substances. In addition, drugs and their metabolites enter the environment after use by patients. The problem of safe collection and disposal of drugs in civil circulation is also relevant. The complex of environmental problems associated with drugs is the focus of international organizations such as the Organization for Economic Co-operation and Development (OECD) or the United Nations Environment Programme (UNEP). Expert groups within their framework assess the situation with environmental pollution by drugs as threatening. The problem of drug pollution requires the use of an approach covering the entire life cycle of drugs – from development and registration to consumption and disposal. The focus should be on pollution prevention measures: developing "green" drugs, limiting the use of substances with high environmental risks, creating drug disposal programs, upgrading treatment facilities, and educational campaigns.

Text. The objective of this review is to conduct a comparative analysis of environmental risk assessment methods used in the European Union (EU), the United States, and Russia. The EU uses a two-stage assessment system (primary screening and detailed analysis) regulated by the European Medicines Agency (EMA). In the United States, the Food and Drug Administration (FDA) applies a categorical exclusion mechanism, which makes the system more flexible. The current environmental legislation in Russia regulates waste management and sets standards for the impact on the environment, while there is no comprehensive risk assessment of the life cycle of drugs.

Conclusion. The use of international experience, including the phased implementation of regulatory requirements for the assessment of environmental risks of drugs, can contribute to the development of a more sustainable pharmaceutical industry in Russia.

On February 17, the annual international congress "Drug Development and Registration" was held in Moscow, organized by the scientific-practical journal of the same name, the Center of Pharmaceutical Analytics, and the Saint Petersburg State Chemical and Pharmaceutical University as co-organizer.Since 2025, a specialized section for young specialists in pharmaceutical science and practice – the "Youth League of Drug Development and Registration" – has been held within the congress. This year the Youth League took place for the second time, but already gained international status and brought together young scientists from the EAEU countries: the Russian Federation, the Republic of Belarus, and the Republic of Kazakhstan. The main goal of the Youth League is to provide young scientists with an opportunity to present at one of the largest events in the pharmaceutical industry alongside leading scientific and practical specialists, representatives of major enterprises, universities, and regulatory authorities. The Youth League is one of the main formats of support provided by the Center of Pharmaceutical Analytics to young scientists conducting research in pharmaceutical chemistry, drug development, preclinical and clinical studies, and many other areas that are the main scientific directions of publications in the journal "Drug Development and Registration", which publishes many articles by young scientists.The main prize, which by tradition has become a certificate for publishing an article in our journal, was contested this year by representatives of the Saint Petersburg State Chemical and Pharmaceutical University, I. M. Sechenov First Moscow State Medical University, Voronezh State University, Ryazan State Medical University named after academician I. P. Pavlov, Perm State Pharmaceutical Academy, Scientific Centre for Expert Evaluation of Medicinal Products, Belarusian State Medical University (Minsk), South Kazakhstan Medical Academy (Shymkent), as well as the Center of Pharmaceutical Analytics. A professional team of experts duly evaluated the participants' works, and in tight competition, the prizes were distributed as follows:Winner: Anna Yu. Grishina (SPCPU, St. Petersburg, scientific supervisor Cand. Biol. Sci. D. Yu. Ivkin).Silver prize: Victoria A. Terletskaya (BSMU, Minsk, scientific supervisor Cand. Pharm. Sci. R. I. Lukashov).Bronze prize: Yulia D. Yatskovskaya (CPhA, Moscow, scientific supervisor Dr. Pharm. Sci. T. N. Komarov).Also, the CPhA team awarded a special prize in the form of a certificate for an internship at the company to Ulyana A. Efremova (SPCPU, St. Petersburg) and Ainur N. Karabaeva (SKMA, Shymkent).Anna V. Schastnaya (BSMU) received a special prize from the journal "Herbarium" by decision of the editorial board.The main reward for us, the organizers, was the fascinating reports of the participants, lively discussion, shining eyes and wonderful atmosphere of the event. We look forward to seeing you at the third "Youth League of Drug Development and Registration" in 2027!This year, in addition to oral presentations, the event also included publication of abstracts in a separate section of our journal, which you can find below.

Разработка и регистрация лекарственных средств. 2025;14(4):174–182. https://doi.org/10.33380/2305-2066-2025-14-4-2207 Статья опубликована: 28.11.2025.