The development of a rational dosage form for an API is one of the main links in the chain of comprehensive research to create a drug. The purpose of this study was to use physicochemical and technological methods to create various parenteral forms based on the practically water-insoluble API LCS-1269 and to conduct a comparative study of the developed dosage forms according to the main quality indicators characteristic of injectable dosage forms. Compositions and technologies for producing three model dosage forms based on LCS-1269 were evaluated during the work. Their main advantages and disadvantages were shown.

Background. Lipophilicity is a fundamental physicochemical property that determines the solubility and transport of a drug through biological membranes, as well as its behavior in the body. Lipophilicity also affects the ability of a drug to bind to plasma proteins and reach the corresponding receptors. The standard for drug lipophilicity experimental study is measuring the distribution between two immiscible phases – aqueous (water and buffer solutions) and hydrophobic (most often octanol).Aim. The experimental study of somatostatin analogue cyphetrylin lipophilicity in octanol/water system distribution test.Materials and methods. The lipophilicity of cyphetrylin, synthesized in the Laboratory of Chemical Synthesis of the N.N. Blokhin National Medical Research Center for Oncology of the Russian Ministry of Health, was studied in the octanol/water system; ethanol was used to study cyphetrylin spectral characteristics and its quantitative determination; shake flask method, UV-spectrometry.Results. The experimental assessment of cyphetrylin lipophilicity was carried out by shake flask method in a system of mutually saturated water and octanol 1:1. Since cyphetrylin is practically insoluble in water, the concentration of the drug in the octanol phase was determined by UV-spectrometry and the concentration in water was calculated by mass balance. Lipophilicity was expressed as the decimal logarithm of the concentration of cyphetrylin in the octanol phase to its concentration in the aqueous phase (logPo/w) ratio. The experimentally determined value of logPo/w was 1.14.Conclusion. The lipophilicity of cyphetrylin was studied experimentally by shake flask method as a parameter that determines the molecule probability to reach the biological target. The logPo/w value 1.14 in decimal logarithmic form indicates moderate lipophilicity of cyphetrylin, which exhibits antitumor activity when interacting with somatostatin receptors.

Modern methods of analytical characterization and quality control of liposomal pharmaceutical products are reviewed. Analytical studies are shown to be extremely important not only for the development and quality control of liposomes but also for understanding the possible clinical safety and efficacy of liposomes with encapsulated drugs. The complicated analytical characteristics of liposomes that are determined by their unique structure, nanoscopic dimensions, and excipients, primarily lipids, are highlighted. Critical quality attributes of liposomes, e.g., qualitative and quantitative determination of the lipid components of the liposomal shell, nanoparticle morphology, encapsulation efficiency, and drug release are discussed. Available analytical approaches and determination methods are summarized. Examples of analytical studies of liposomes with encapsulated anticancer drugs are given.

The indolocarbazole derivative LCS-1269 has a multitarget mechanism of action on tumors and blocks vasculogenic mimicry. The hydrophobic character of LCS-1269 hampers the development of an injectable dosage form (IDF), the need for which was determined in series of in vitro and in vivo biological experiments. The purpose of this study was to select an excipient composition for creating a model IDF of LCS-1269 with high antitumor activity and low toxicity and to develop the technology for LCS-1269 IDF production. Acomposition of a model IDF of LCS-1269 including EtOH (95%), Kollidon 17 PF, and polysorbate 80 and the technology for obtaining a true solution were developed during preliminary experimental studies. The selected model was lyophilized to exclude EtOH from the composition and to increase the shelf life of the compound.

Epidermal growth factor receptor (EGFR) inhibitors are widely used in the treatment of lung cancer, one of the most common malignancies. Gefitinib, an EGFR tyrosine kinase inhibitor, has limited clinical efficacy due to slow absorption in the gastrointestinal tract and low and variable bioavailability after peroral administration. The purpose of this study was to develop a quantitative determination method for gefitinib in a liposomal dosage form intended to increase its bioavailability and therapeutic efficacy. Direct UV spectrometry was used for this. The developed methodology was validated in accordance with current recommendations. The liposomal bilayer components were shown not to affect the spectral characteristics of gefitinib and did not interfere with its quantitative determination, i.e., the technique was specific. Validation confirmed the linear dependence of the analytical signal on the gefitinib solution concentration. The method provided acceptable accuracy and precision in the analytical range from 70 to 130% of the nominal gefitinib content in a unit dosage form. The results were not burdened by systematic error. The limits of detection and quantitation calculated using the linearity parameters were 0.26 mg and 0.80 mg of gefitinib, respectively. The extended analytical range of the technique allowed it to be used to quantify and determine the dosing uniformity of gefitinib in the lyophilized liposomal dosage form.

Производное индолокарбазола ЛХС-1269 обладает мультитаргетным механизмом воздействия на опухоль и блокирует васкулогенную мимикрию. По своим свойствам ЛХС-1269 относится к гидрофобным соединениям, что затрудняет получение инъекционной лекарственной формы (ИЛФ), необходимость создания которой определена в ходе ряда биологических опытов in vitro и in vivo. Цель настоящего исследования: выбор композиции вспомогательных компонентов для создания модели ИЛФ ЛХС-1269 с высокой противоопухолевой активностью и низкой токсичностью и разработка технологии ее получения. В ходе предварительных экспериментальных исследований разработаны состав модели ИЛФ ЛХС-1269, включающий спирт 95 %, Kollidon 17PF и полисорбат 80, и технология, позволяющая получить истинный раствор ЛХС-1269. Для исключения спирта из состава и для увеличения срока годности проведена лиофилизация выбранной модели.

Лапатиниб одобрен Министерством здравоохранения РФ в качестве препарата 1 линии терапии наиболее агрессивной формы рака молочной железы (РМЖ), связанного с гиперэкспрессией рецептора эпидермального фактора роста 2-го типа (Human epidermal growth factor receptor 2; ErbB2 (HER2/neu)). Режим дозирования препарата предполагает прием от 4 до 6 таблеток лапатиниба в сутки в течение как минимум 14 дней. В условиях нарушения цепочек поставок препаратов на территорию РФ на первый план выходит создание эффективного и безопасного воспроизведенного препарата (дженерика) для беспрерывного обеспечения им пациентов. Целью данной работы являлось проведение комплексного исследования по изучению возможности использования прямого прессования для получения воспроизведенного лекарственного препарата Лапатиниб (таблетки, покрытые пленочной оболочкой 250 мг). В состав таблетки оригинального препарата входит около 42 % микрокристаллической целлюлозы, которая используется как наполнитель и связующее вещество для улучшения текучести смеси для таблетирования и повышения прочности таблетки. Это указывает на возможность применения прямого прессования, как наиболее простого и доступного метода. В ходе исследования показано, что смесь для таблетирования, воспроизведенная по прописи оригинального препарата, вне зависимости от способа смешения ингредиентов не обладает необходимыми технологическими характеристиками для получения таблеток методом прямого прессования, несмотря на хорошую прессуемость смеси и прочность полученных таблеток (прочность на раздавливание 80 H). Работа по воспроизведению таблеток лапатиниба будет продолжена с использованием метода влажного гранулирования. Lapatinib is approved by the Ministry of Health of the Russian Federation as a first-line drug for the most aggressive form of breast cancer (BC) associated with overexpression of the Human epidermal growth factor receptor 2 (ErbB2 (HER2/neu)). Patients need to take 4 to 6 lapatinib tablets per day for at least 14 days. In the context of a disruption in the supply chains of drugs to the territory of the Russian Federation, it is necessary to create an effective and safe generic drug for the continuous provision of patients with it. The purpose of this work was to conduct a comprehensive study to discover the possibility of using direct compression to obtain a generic drug Lapatinib, film-coated tablets 250 mg. The tablet of the original drug contains about 42% microcrystalline cellulose, which is used as a filler and binder to improve the fluidity of the tablet mass and increase the strength of the tablet. This indicates the possibility of using direct pressing as the simplest and most accessible method. The study showed that the mixture for tableting, reproduced according to the prescription of the original drug, regardless of the method of mixing the ingredients, does not have the necessary technological characteristics for obtaining tablets by direct compression, despite the good compressibility of the mixture and the strength of the obtained tablets (diametrical crushing strength of 80 H). Work on reproducing lapatinib tablets will continue using the wet granulation method.

Liposomes are one of the most well-known and promising nanoscale drug delivery systems. Liposomal medicinal products are successfully used in clinical practice for cardiovascular, oncological, dermatological, and other indications. The development of liposomes and their widespread implementation in clinical practice are relevant tasks. The aim of the study was to summarise and analyse scientific data regarding the structure, composition, and classification of liposomes, as well as specific aspects of liposome production methods. This review covers up-to-date information on marketed liposomal medicinal products. The authors illustrate how production methods affect the composition and structure of liposomes. The size and lamellarity are important characteristics of liposomes that determine the encapsulation efficiency and biodistribution of active pharmaceutical ingredients (APIs). The choice of excipients depends on the intended use of liposomal medicinal products. The article summarises the main liposome production methods, highlighting specific usage aspects, advantages and disadvantages. Conventional liposome production methods are easy to apply and do not require complex equipment, and their principal disadvantages include the low efficiency of API encapsulation within liposomes and the high complexity of scaling up technological processes. The authors pay special attention to microfluidic techniques for liposome preparation, which are characterised by a highly controlled technological process (in terms of size distribution and lamellarity), high reproducibility, and scalability to the level of industrial production and are applicable to encapsulating different APIs.

The N.N. Blokhin Oncology Research Center synthesized indolo[2,3-a]carbazole N-glycoside, which received the laboratory cipher LСS-1269, which showed a pronounced antitumor effect during research.. During the study of LСS-1269 by HPLC, the presence of two impurities was revealed, called LСS-1269-X and LСS-1269-Y (impurities X and Y). The electronic absorption spectrum (EAS) of the impurity peak X was similar to the EAS of the indolo[2,3-a]carbazole N-glycoside obtained and described earlier under the laboratory cipher LCS-1208, and the EAS of the impurity was the spectrum of LCS–1269, which indicates that the impurities belong to indolo[2,3-a]carbazoles. In this regard, it is suggested about the structure of these impurities: impurity X is a xyloside of aglycone similar in structure to aglycone LCS-1208, which can be formed as a by-product at one of the last stages of the synthesis of LCS-1269, and impurity Y has the structure of triacetyl ester LCS-1269, which is a precursor of LCS-1269 in the synthesis process. The aim of the work was to identify impurities in the substance LHS-1C69, to confirm their structures and belonging to derivatives of indolo[2,3-a]carbazole. The identification of impurities in the substance LCS-1269 was carried out using the HPLC-MSMS method. For this purpose, X and Y impurities having the assumed structures were obtained by directed counter synthesis, which were used as reference samples. Comparison of peak retention times on extracted ion chromatograms (XIC) and MSMS spectra of impurities with XIC and MSMS spectra of samples of comparison of the alleged impurities confirmed their structure. As a result of the work carried out, the structures of impurities X and Y were identified and the affiliation of these impurities to derivatives of indolo[2,3-a]carbazole was confirmed.

Background. Expression evaluation of somatostatin receptors (SSTRs) in tumor cells is necessary for the reasonable use of therapy directed at such receptors.Aim. The affinity determination of the original analogue of somatostatin cyphetrylin for SSTRs of transplanted mice mammary adenocarcinoma Ca-755.Materials and methods. Cyphetrylin was synthesized in the Chemical Synthesis Laboratory of the Research Institute of Experimental Diagnostics and Therapy of Tumors, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of Russia. Cyphetrylin in tablet form was administered orally at a therapeutic dose of 10 mg/kg for 7 days to female F1 (C57Bl/6 × DBA/2) tumor-grafted Ca-755 mice. Animals of the control group were not administreted with сyphetrylin. Tumor tissue samples were obtained from animals on the 9th and 14th days after Ca-755 transplantation and sent for immunohistochemical study, which was performed on serial paraffin sections by the immunoperoxidase method using primary antibodies to various types of SSTRs.Results. The high frequency of positive expression of SSTR1, SSTR2, and SSTR5 (in 80, 100 and 100 % of tumor samples, respectively) was shown in tumor samples of the control group animals. As a result of cyphetrylin introduction in tumor samples obtained on the 9th day after Ca-755 inoculation, a change in the tumor receptor status was found towards a decrease in the level of expression of SSTR2 (80 % of samples) and SSTR5 (80 % of samples); SSTR1 expression did not change (80 % of samples). Compared to the controlled, in tumor samples after cyphetrylin administration, obtained on day 14 from Ca-755 transplantation, a decrease in the expression level of SSTR2 (80 % of samples), SSTR1 and SSTR5 (60 % of samples for SSTR each type) was noted, due to cyphetrylin binding to SSTRs of tumor cells. The receptors SSTR3 and SSTR4 did not show a high level of expression in the studied Ca-755 tumor samples.Immunohistochemical staining of Ca-755 cells with antibodies to SSTRs showed a tendency to reduction of antigen-positive cells number from 15–50 % in control to 10–40 % on day 9 after Ca-755 transplantation and 10–30 % on day 14 after Ca-755 transplantation.Conclusion. The data obtained indicates the presence in mice transplanted mammary adenocarcinoma Ca-755 of SSTR1, SSTR2 and SSTR5 high level expression due to the binding to which the direct cyphetrylin antitumor effect is realized.

Lapatinib is a small molecule, a heterocyclic quinazoline derivative. The drug is used for targeted therapy of patients with breast cancer, in which there is overexpression of the human epidermal growth factor receptors (HER/ErbB). This review is devoted to studying modern instrumental methods of qualitative and quantitative analysis of lapatinib, which can be used both for quality control and standardization (of bulk pharmaceuticals and dosage forms) and pharmacokinetics studies of a drug. Reverse-phase high-performance liquid chromatography (RP-HPLC) is mainly used to identify lapatinib in tablets. Depending on the purpose of the study, various detectors are used (ultraviolet or diode-matrix detector), which makes it possible to determine not only the native compound but also the products of its degradation. Definition of lapatinib in the presence of degraded products is necessary for forced degradation studies to determine drug stability. When a drug is being developed, it is important to define and understand its pharmacokinetics. For such studies, high-performance liquid chromatography (HPLC) coupled with the mass selective detector is often used. It allows determining lapatinib in biological fluids. However, these methods are not applicable for identifying the drug directly in dosage forms and require further development and validation.

Background. A glycoside derivative of indolocarbazole LHS-1269, one of the new drugs selectively affecting tumors, which was first synthesized at the N.N. Blokhin National Medical-Research Center of Oncology of the Ministry of Health of Russia, is of particular scientific interest. Experimental studies demonstrated a multi-target mechanism of action of this compound. LHS-1269 interacts with several intracellular targets and induces various pathways of cell death. Several innovative models of the dosage forms were designed to achieve the highest antitumor activity of the compound and to perform further preclinical studies. Aim. To develop the methods for the quantitative determination of LHS-1269 in pharmaceutical compositions proposed as a result of the search for the optimal dosage form. Materials and methods. The study analyzed the spectrophotometric characteristics of LHS-1269 solutions in dimethylformamide, dimethylsulfoxide (DMSO) and in the mixture of solvents DMSO–ethyl alcohol, as well as electronic absorption spectra of the excipients in the mixture of solvents DMSO–ethyl alcohol. Spectrophotometric measurements were performed on a Cary 100 spectrophotometer (Varian, Inc., Australia) in the wavelength range from 200 to 500 nm. The standard sample is the substance LHS-1269 (N.N. Blokhin Oncology Research Center of the Ministry of Health of Russia). Results. The carried out studies showed that LHS-1269 solutions in dimethylformamide, DMSO and mixture of DMSO– ethyl alcohol are suitable for spectrophotometric measurements. Several variants of the methodology for the assay of LHS-1269 in various dosage form models that differ in the content of the active substance and the excipients composition have been developed: LHS-1269 concentrate for solution for injection and infusion; lyophilisate for solution for injection; liposomal lyophilisate for dispersion for injection. Conclusion. Techniques for the assay of LHS-1269 in dosage form models have been developed. It has been shown that the developed techniques are applicable for LHS-1269 quantitative determination in innovative dosage forms containing polymeric low molecular weightsolubilizers, lipids, cholesterol, mono- or oligosaccharides as excipients.

Neuroendocrine tumors (NET) is a rare pathology characterized by dysfunction of the pituitary gland and endocrine glands. The survival prognosis depends on the stage of the disease at which the malignant growth was detected.The study objective was to summarize information on the methods of early diagnosis of NETs and antitumor therapy using peptide hypothalamic hormone somatostatin analogues.The review provides a chronological analysis of studies of the expression of somatostatin receptors (SSTRs) in different tissues and organs, methods of their visualization and quantitative determination over 50 years. The expression of SSTRs was revealed on NET cell membranes. The history of SSTRs study dates back to 1972, when somatostatin was first isolated and its structure established. Later, the peptide therapeutic activity in the treatment of NET, realized through somatostatin receptors, was discovered, and a number of somatostatin analogues were synthesized. Five main types of SSTRs are known, including subtypes which are expressed in characteristic volumes in the corresponding pathologies. Determining the expression of SSTRs provides diagnosis and staging of malignant neoplasms. Now the regulation of tumor growth is carried out by synthetic somatostatin analogues. SSTRs imaging is an important stage in planning NET therapy with drugs of this pharmacological group. Clinical studies of the therapy of tumors of various organs and tissues, in vitro and in vivo imaging are being carried out, methods for detecting SSTRs are being studied and compared.A retrospective analysis of the methods used to visualize SSTRs and diagnose and treat NETs is presented. It has been shown that positron emission tomography combined with computed tomography, using SSTRs-binding radioligands is the safest and most accurate method for visualizing SSTRs and diagnosing NETs.

Liposomal targeted drug delivery makes it possible to achieve effective concentration in the target cell under various pathological conditions. The main advantage of liposomal particles is their biodegradability and immunological neutrality, which improves the safety profile of drugs. The review provides information on the composition of liposomes: the main component of the liposomal membrane is phospholipids, which provide its strength and protect from mechanical impacts. Liposomal particles are distinguished by the size and number of bilayer membranes, also secreted liposomes with a non‑lamellar organization. The composition and size of liposomes are selected depending on the purpose, including excipients in the membrane that affect the properties and functions of liposomes, including the rate of release of the components, the affinity of liposomes for the target tissue, etc. The review considers the main methods for obtaining liposomes and the features of their use, advantages and disadvantages. The creation of liposomes that are sensitive to various external or internal physicochemical factors makes it possible to realize drugs effects, localize the site of its action and reduce the number and severity of side effects. Currently, liposome‑based drugs are successfully used in various fields of medicine – dermatology, cardiology, oncology, neurology, etc. The most active condact preclinical and clinical studies of liposomal drugs for the treatment of malignant neoplasms. Particular attention is paid to the work of Russian researchers in the field of targeted drug delivery. It is shown that today liposomes are an open for study and improvement system for targeted drug delivery.

The primary purpose of the present study was to apply the quality by design approach to the formulation of the somatostatin analog (SSA) cyphetrylin dosage form.Experimental investigations of cyphetrylin activity in vivo showed that its oral administration is optimal.For better patient compliance and product quality, an interdisciplinary team identified the quality target product profile and determined the critical quality attributes (CQAs) related to product safety and efficacy.On the basis of our own in vitro/in vivo nonclinical data and experience of SSA medicinal use, we estimated the criticality of each CQA by a specially created scale.Because of the minimal quantity of the active substance in the tablet, the assay and uniformity of content are defined as the most CQAs.Estimation of the manufacturing process by methods of the risk analysis and mitigation matrix and failure mode effect analysis allowed us to determine the effects of process unit operations on final product in-process CQAs and to characterize the granulation and compression as the most critical steps.It has been shown that the moisture content of granulate is the most significant in-process CQA, which affects the disintegration and resistance to crushing of tablets and their microbial limits.

Introduction. Indolocarbazole derivatives are of increasing scientific interest for practical oncology. A number of N-glycosides, indolo[2,3-a] carbazole under the laboratory code LCS, were synthesized in the laboratory of chemical synthesis of the National Medical Center of Oncology named after N.N. Blokhin. Currently, one of the most promising compounds in this class is LCS-1208, a representative of the arabinoside class of indolo [2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione. According to the mechanism of biological action, LCS-1208 is a protein kinase C inhibitor and is of great interest for the treatment of malignant neoplasms.Aim. chemical and pharmaceutical standardization of the pharmaceutical substance LCS-1208.Materials and methods. Laboratory samples of pharmaceutical substance LCS-1208. Methods of investigation: gravimetry, spectrophotometry, polarimetry, high-performance liquid chromatography (HPLC), high-resolution nuclear magnetic resonance (NMR) spectroscopy and infrared (IR) spectroscopy.Results and discussion. The quality assessment of LCS-1208 was carried out according to the indicators adopted in the XIV edition of the State Pharmacopoeia of the Russian Federation for quality control of pharmaceutical substances. LCS-1208 - orange amorphous powder, odorless; soluble in dimethylsulfoxide (DMSO) and dimethylformamide (DMF); very slightly soluble in 95 % ethyl alcohol and practically insoluble in water. The authenticity of the substance is confirmed by NMR and IR spectra, as well as electronic absorption spectra. The values of the specific optical rotation of LCS-1208 (1 % solution in DMF) are placed in the range from +58° to +61°. All the studied samples of the substance were free of inorganic impurities, sulphate ash, heavy metals and contained no more than 1.0 % water, determined by the K. Fischer titration method. The content of possible related impurities in the substance LCS-1208 and the content of the main active substance were determined by HPLC. The studied laboratory series of the pharmaceutical substance LCS-1208 contained no more than 1.0 % of any single and no more than 3 % of the total unidentified impurities. The content of the main active substance was more than 97 %.Conclusion. As a result of the work carried out, quality criteria and parameters were selected and methods for their determination were developed, which allow to adequately assess the quality and standardness of the pharmaceutical substance LCS-1208.