Vol 18, No 4 (2020)

Progress in clinical research and medicine has contributed to a significant reduction in the worldwide mortality rate. Currently, the strategy of developed countries is directed at increasing expectancy and improving life quality. As a consequence, the modern world is already faced the problems of aging and age-related diseases. Neurodegenerative diseases are one of the main medical problems suffered by an «aging» population of people. Stem cells as a part of biomedical cell products (preparations based on viable somatic human cells), which can have a regenerative function on the brain and spinal cord neurons or can be used for replacement therapy, are considered as one of the directions of the development of new approaches to the treatment for neurodegenerative diseases. The paper describes the features of using and development of stem cell-based drugs promising for the treatment of such neurodegenerative diseases as multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, and lateral amyotrophic sclerosis. There are considered clinical studies conducted in the world with the use of mesenchymal stem cells from umbilical cord blood, bone marrow, neuronal and embryonic stem cells for the treatment of neurodegenerative diseases. Currently, clinical studies of preparations based on viable human cells for the treatment of neurodegenerative diseases are mainly characterized by the proven safety of their use and specific immunomodulatory activity. Given at the early stages of clinical trials involving a limited number of patients mainly in single-center localization, data on the efficiency of using the stem cells regenerative potential for the treatment of neurodegenerative diseases will be available in the future.

The problems of longevity increase and its quality improvement are actual for the modern gerontology. So far it is necessary to complete tasks of early diagnostic and target prophylactic of the age-associated pathology. There are presented the data concerning β-microglobulin (β2М), one of the frequent marker of aging. There are considered the data about the structure, properties, and functions of β2М, in which concentration is increased by aging. The experiments demonstrated the β2М concentration to increase in the hippocampus of old animals as in young heterochronic parabionts. The great correlation is observed between the β2М concentration and the deterioration of cognitive functions as well as the development of dementia. β2М plays a great role in the development of cardiovascular disease. The heart ischemic disease, the pathology of arteria carotis and peripheral vessels were more expressive in the patients with a great β2М concentration in plasma. It was the correlation between diabetes mellitus severity and high β2М level. β2М, as an oncogenic factor, stimulates the development and metastasis of the different cancers. The determination of plasma β2М concentration is of great significance for the predictive medicine and the prophylactic of the age-associated pathology.

Introduction. To assess the pharmacokinetic indices of new drugs, it is necessary to develop analytical methods for determining their concentration in biological objects. Purpose of the study. Development and validation of HPLC-mass spectrometric methods for the determination of a new valproic acid derivative and 1,3,4-thiadiazole in rat brain. Methods. The object of the study was a new anticonvulsant N- (5-ethyl-1,3,4-thiadiazol-2-yl)-2-propylpentanamide (valprazolamide). Determination of valprazolamide was carried out by the HPLC-mass spectrometric method. Selectivity, sample transfer, linearity, accuracy, precision, matrix effect, the degree of extraction of valprazolamide were evaluated. Results. A method of HPLC-mass spectrometric determination of valprazolamide in rat brain homogenates was developed (Zorbax Eclipsi plus C18 analytical column - 4 μm 2,0*150 mm, temperature - 30°C; the mobile phase is acetonitrile and deionized water with the following gradient profile: 0-1 minute 10% acetonitrile; 1-5 minutes a linear increase in the concentration of acetonitrile to 90%; 5-6 minutes isocratic plot with a concentration of acetonitrile 90%; 6-10 minutes column conditioning with 10% acetonitrile; the flow rate of the mobile phase is of 0.6 ml/min; the volume of the injected sample accounts for 10 μl; total elution time - 10 minutes, mass detection with negative polarization). The MRM transition of valprazolamide amounted to m/z 254,0 → m/z 167,1. Conclusion. The analytical limits of the method are in the range from 1 to 1000 ng/ml. The developed method is selective, accurate, precise and linear, it meets the requirements for the validation of bioanalytical methods in all respects.

Introduction. The specific activity of blood coagulation factor VIIa depends, among other things, on the presence of N-acetylneuraminic acid (sialic acid) and phosphate residues in the molecule. Thus the authenticity of glycosylation profile as one efficacy and safety proofs of drug product is of particular interest. However, the whole variability of posttranslational modifications requires an individual approach to corresponding assessment processed in accordance with the requirements and recommendations of the comparison of biosimilar drug products. Methods. Objects under the study referred as: original drug product NovoSeven, biosimilar drug product Coagil-VII and European reference standard of Human Coagulation Factor VIIa (rDNA) CRS. Two methodological approaches of high-pH-anion-exchange chromatography (HPAEC) and hydrophilic interaction liquid chromatography (HILIC) with fluorescent and mass-spectrometric detection were used for the analysis of oligosaccharide distribution in all factor VIIa molecules. Results. А set of rFVIIa modifications represented basically by complex biantennary oligosaccharides with insignificant part of triantennary structures was established in this study. The most characteristic modification for Coagil-VII biosimilar was complex bisialylated core-fucosylated biantennary group. Conclusion. The evaluation of the N-glycosylation profile of recombinant blood coagulation factor VIIa based on the comparative analysis of biosimilar drug product, original drug product, and CRS standard sample revealed difference in quantitation of N-glycans among with general matching of the majority of peak retention times. Mass-spectrometric detection allowed to carry out a structural interpretation of HPLC data. A biosimilar drug product was used as an example to illustrate the necessity for factory standards certified in a prescribed manner for the further glycan profile authentication.

Introduction. The mainfunction of alpha-1 antitrypsin (A1AT) is inhibition of neutrophil elastinase and control over TNFα secretion. Anti-inflammatory properties of A1ATare manifested in leucocyte migration decrease and proliferation of T-lymphocytes through an increase in the amount of cAMP in the cells. Study objective. To examine the level of A1AT in chronic lymphocytic leukemia (CLL) patients at different stages of the progression of the main disease, as well as depending on the type of infectious complications and location of the pathological process. Methods. Data of 177 CLL patients in dynamics were analyzed. Methods for examining etiologic agents of CLL infectious complications were applied according to Order No.535 «On Unification of Microbiological (Bacteriological) Examination Methods, Applied in Treatment and Diagnostic Laboratories of Medical and Preventive Institutions» dated April 22, 1985. ELISA method was applied to identify antibodies to the viruses Varicella zoster, Cytomegalovirus. A1AT concentration also was measured with ELISA method with the use of test systems IDKa1-Antitrypsin Clearance ELISA (Immundiagnostik AG company, Bensheim). Results. The examination of the A1AT concentration mostly showed its moderate increase under bacterial infections. A significant increase in the A1AT content with respect to the normal values was identified under viral and mycotic complications. Significant difference between the serum A1AT level under the treatment of infectious complications and their base values was identified (р<0,001): after the beginning of the treatment of infectious complications, the A1AT level increased by 24,7% under bacterial complications, by 9,8% under viral complications and by 7,0% under mycoses. The A1AT level became 1,5-2 times lower during the stabilization of the process and leveling of clinical implications of the infection. Conclusion. The mechanism of the А1АТ level increase under the effect of the medications may be associated both with the death of infectious agents, the destruction of which causes release of pyrogenic agents and endotoxins, triggering the mechanisms of inflammatory reaction development and causing significant release of А1АТ in patients with CLL, and with a hepatotoxic effect of chemotherapy. Therefore, measurements of the A1AT level can be used to control effectiveness of treatment of infectious complications in patients with CLL.