PREPARATION, CHARACTERIZATION AND STUDIES OF PHYSICOCHEMICAL AND BIOLOGICAL PROPERTIES OF DRUGS COATING LACTOSE IN FLUIDIZED BEDS

Objective: Study physicochemical properties and activity of biotechnological drugs coating lactose particles in fluidized beds for the development of a prospective approach of their identification. Methods: Lactose monohydrate as pharmaceutical excipient; affinity-purified polyclonal rabbit antibodies to recombinant human interferon-gamma as a drug substance; Pilotlab fluid bed apparatus was used for lactose powder saturation with solutions of pharmaceutical substances to the point of granulation (pelletizing); inverse light scattering method (2D-LS) for analysis of micron vibrations frequency spectra of samples surfaces for light intensity distribution in time by values of d1, d2, d3 primary descriptors; low angel and dynamic laser light scattering (LALLS, DLS) methods for distribution of lactose-water (LW) supramolecular complexes into volume fractions (micron "size spectra"), using the Master Sizer 2000 instrument and Zeta Sizer Nano ZS instrument in the nanoscale; Spirotox method for research of biological activity to determine the activation energy (Ea) values of cell death in solutions of tested samples. Results: Changes in 2D-LS scattering time on sample surfaces, described by topological descriptors, made it possible to clearly differentiate the intact lactose from fluidized samples by specific corridors in coordinates di=F(t). The calculated activation energy (Ea) values of cell biosensor death process in solutions of drugs coating lactose allow to characterize the biological activity of it in the initial (by Ea increase) and activated state (by Ea decrease) after the creation of intra-laboratory transmucosal conditions. A unique dimensional spectrum of LW complexes in the nanoscale range was obtained by DLS. The differences between samples in the distribution of LW complexes in the size range from 1 µm to 125 µm was showed by LALLS. Conclusion: The developed approach, including Сhemometrics, laser and biotesting methods can be used for qualitative the analysis tasks as well as for analytical control of the fluidization process in cases where identifiable pharmaceutical substances are not distinguishable by traditional analytical methods

О биологической активности и супрамолекулярной структуре морских вод: проблемы использования природных вод в составе лекарственных средств

Modern approaches to definition of properties and qualities of the natural water intended for manufacturing of pharmaceutical preparations are considered. It is shown, that waters of the internal seas render toxic influence on a cellular biosensor control that is connected with significant anthropogenous loading on them. The analysis of water supramolecular structure can serve the express train-estimation of quality of suitability of water for application in the medical purposes.Рассмотрены современные подходы к определению свойств и качества природных вод, предназначенных для изготовления фармацевтических препаратов. Показано, что воды внутренних морей оказывают токсическое воздействие на клеточный биосенсор, что связано со значительной антропогенной нагрузкой на них. Анализ супрамолекулярной структуры воды может служить экспресс-оценкой качества пригодности воды для применения в медицинских целях

LIGHT SCATTERING IN RESEARCH AND QUALITY CONTROL OF DEUTERIUM DEPLETED WATER FOR PHARMACEUTICAL APPLICATION

Objective: Development of a methodology for measuring the deuterium content in water for pharmaceutical purposes by laser light scattering based on ideas about the cluster structure of water. Methods: Samples of industrially manufactured drinking water from different manufacturers with varying deuterium content from 10 ppm to 115 ppm. For the titration of laboratory samples of deuterium depleted water in increments of 5 ppm the following reagents were used: Water, deuterium-depleted (≤1 ppm (D2O, Aldrich, USA); Deuterium oxide/Heavy water/Water-d2 (99.9 atom % D, Aldrich, USA); water Milli-Q (specific resistance 18.2 µS·sm at 25 оС, ТОС ≤ 5 ppb, Merck Millipore). The determination of deuterium content in samples of industrially manufactured water and water obtained in a laboratory manner was carried out by the method of low-angle laser light scattering (LALLS) at the Mastersizer (Malvern Instruments) analyzer and using a working measuring tool–laser dispersion meter/MDL («Cluster-1», Russia/Ukraine). The statistical methods–packages OriginPro®9. Results: It was found that the content of isotopologies in water leads to physicochemical water’s properties changes and morphology changes of giant heterogeneous clusters (GHC). The results of low-angle laser light scattering (LALLS) in the water samples under investigation showed the dependence of the water GHC "dispersibility" expressed in the differentiation of curves of the volume size distribution ("size spectra"), the volume concentration, w%, the laser obscuration values (I ‒I0) as the function of the water isotopic composition variations. The laser diffraction method results correlate with two-dimensional (2D) multi-descriptor mathematical analysis. Conclusion: When identifying deuterium depleted water, it should be considered not only the indicators that determine its pharmacopoeial quality, but also the D/H ratio, because even small changes in the natural isotopic composition of water lead to significant biological effects. Our proposed approach using laser diffraction in combination with mathematical apparatus of (2D) multi-descriptor laser scattering analysis makes possible the exact calculation of individual signs of deuterium depleted water as the pharmaceutical object of study

POLARIMETRY AND DYNAMIC LIGHT SCATTERING IN QUALITY CONTROL OF CARDIOTONIC AND HYPOTENSIVE TINCTURES

Objective: To substantiate the possibility of using polarimetry to control the quality of tinctures as an additional pharmacopoeial method. Methods: The polarimetric method (POL-1/2, Atago, Japan, the measurement accuracy of±0.002 °) was used to measure the optical activity (α °) of motherwort, valerian and hawthorn tinctures. The dynamic light scattering method (DLS; Zetasizer Nano ZS, Malvern, UK) was used to assess the stability of alcoholic and aqueous dilutions of tinctures according to the intensity of dynamic light scattering dependent on the size (d, nm) of the dispersed phase particles and the values of the electrokinetic potential (ξ, mV). Results: For the first time in this investigation, the polarimetry approach was proposed to evaluate the cardiotonic and hypotensive tinctures' quality and for their identification. Valerian tincture, dilution 1:40,-0.10°<α°<-0.89°; motherwort, tincture-dilution 1:10,-0.10°<α°<-2.21°; hawthorn, tincture without dilution,-0.76°<α°<-1.55°-these are the acceptable ranges of optical activity (α°) of their alcohol dilutions. Beyond these intervals, the use of the polarimetric approach is impossible. Values of optical activity below 0.1 correspond to too low a content of optically active components. Tinctures with optical activity above the upper value of the interval were unstable dispersed systems with low values of the electrokinetic potential (|ξ|≪25mV) and micron particle sizes. Reference tinctures were made from raw materials (Leonurus cardiaca L.) to verify the results. The quality parameters: optical activity (α°), spectra of dynamic light scattering by intensity, volume, and number ("I-d"; "V-d"; "N-d"), electrokinetic potential (ξ) values, and photon pulse count per second (Count Rate, kcps) corresponded to the results obtained for pharmaceutical dosage forms. Conclusion: The permissible intervals of optical activity (α°) of their ethanol dilutions, as well as their relationships with the particle size of the dispersed phase and the values of the electrokinetic potential, were established for the first time to evaluate the quality of tinctures. The obtained results show that polarimetry can be recommended as an additional pharmacopoeial quality control method for tinctures

MECHANOCHEMICAL ACTIVATION OF PHARMACEUTICAL SUBSTANCES AS A FACTOR FOR MODIFICATION OF THEIR PHYSICAL, CHEMICAL AND BIOLOGICAL PROPERTIES

Objective: Study the influence of the mechanical preparation methods (grinding, fluidization) of solid pharmaceutical substances (PS) and herbal raw material on their physicochemical properties and biological activities. Methods: Test substances and solvents-Lactose monohydrate (DFE Pharma, Germany). Sodium chloride, bendazol hydrochloride (all Sigma-Aldrich, USA) and herbal raw material (Callisia fragrans). The dispersity and native structure of pharmaceutical substances were analyzed by several methods: optical microscopy–Altami BIO 2 microscope (Russia); low angle laser light scattering (LALLS) method (Malvern Instruments, UK); Spirotox method–Quasichemical kinetic of cell transition of cellular biosensor Spirostomum ambiguum; Fourier-transform infrared spectroscopy–the analysis in the middle IR region was carried out using an IR Cary 630 Fourier spectrometer (Agilent Technologies, USA). The analysis of dried leaves of C. fragrans before and after mechanical activation was performed using Shimadzu EDX-7000 X-ray fluorescence spectrophotometer without mineralization (Shimadzu, Japan). Results: It was established that the mechanical change, such as dispersion and drying, alters the biological activity of PS and herbal raw materials. The observed increase in the influence of the dispersed substance on the biosensor S. ambiguum is quantitatively estimated from the values of the activation energy (obsEa), which turns to be valued 1,5 (P≤0,05) times more than for the native form substance. In the study of the dependence of the availability of chemical elements K, Ca, Zn on the degree of dispersion of herbal raw materials was established a quantitative 4-fold (P≤0,05) increase in the concentration of elements in mechano-activated raw materials. Conclusion: By the example of the biological model of Spirotox (single-celled biosensor S. ambiguum) and herbal raw materials obtained from C. fragrans, the increase of biological activity of PS at the dispersion of initial preparations was proved

SLOW QUASIKINETIC CHANGES IN WATER-LACTOSE COMPLEXES DURING STORAGE

Objective: To investigate kinetic changes in the spectral characteristics by Fourier Transform Infrared spectroscopy (FTIR) of water-lactose complexes (SMC), derived during the manufacturing process of the drug, containing release-active forms of antibodies. Methods: lactose monohydrate substance, saturated with release-active forms of affinity-purified polyclonal rabbit antibodies to recombinant human interferon-gamma (RA forms of Abs); tablets produced from this substance by direct compression after the addition of excipients (microcrystalline cellulose, magnesium stearate). Powdered and tableted placebo samples saturated with technologically processed water or phosphate-buffered saline, as well as with intact ethanol were used as control. Kinetic changes in SMC were studied using an Agilent Cary 630 FTIR spectrophotometer with a diamond ATR accessory (Agilent Technologies, USA). We used the method of X-ray fluorescence spectroscopy (EDX-7000 Shimadzu energy dispersive X-ray fluorescence spectrometer) to track changes in the fluorescence signal at certain wavelengths. The range of measured elements–11Na-92U. Results: Control of some technological characteristics of the obtained active substance (moisture, flowability) and dosage form (mean mass, disintegration rate) was used as indirect indicators of quality, but they did not allow reliably distinguishing intact lactose from the saturated one. Long-period oscillations on FTIR spectra were characteristic for all types of samples; oscillations occur at approximately two-week intervals; S/N indices were more stable for samples of RA forms of Abs than for placebo samples. On some days, the substance saturated with RA forms of Abs significantly differed from the intact lactose powder. The kinetics of the X-ray fluorescence intensity at certain wavelengths indicates the possibility of a periodic cooperative trigger transition of the system. Reversible conformational transitions are observed for powders on the 30th and 130th days (Kα 3.313 keV). For tablets at Kα 3.313 keV and Kα 1.740 keV small changes were visualized on those days (100–110th day) when hysteresis phenomena were recorded in the IR spectra of these samples. Conclusion: As a result, the evidence for a long-period dramatic conformational mobility of the water-lactose complex was obtained. Based on the data on the semiannual kinetics of IR spectra, a universal criterion for the identity of lactose powder saturated with RA forms of Abs was obtained. Also, it was confirmed that the lactose conformation state was changed by saturation with RA forms of Abs

DEUTERIUM AS A TOOL FOR CHANGING THE PROPERTIES OF PHARMACEUTICAL SUBSTANCES (REVIEW)

The review is devoted to the influence of the hydrogen isotope–deuterium on biological models of organisms and the biological activity of pharmaceutical substances. The positions of the influence of deuterium on the properties of active pharmaceutical ingredients and excipients are examined from different perspectives. The first position reflects an increase in the kinetic isotope effect (KIE) in processes involving known pharmaceutical substances in aqueous solutions with a deuterium/protium ratio (D/H) below natural. For the first time, the dose-response diagram shows the identity of deuterium with essential trace elements, when a deficiency and excess of an element reduces the organism's vitality. Improved kinetic characteristics are demonstrated for the molecular and organism levels of different hierarchical gradations. In particular, they consist in the possibility of increasing the dissolution rate of substances by influencing the carbohydrate mutarotation processes and the optical activity of chiral substances, increased accumulation of essential elements in medicinal plants and other processes associated with a possible change in metabolic pathways in the cell and the organism as a whole. The second considered position of the influence of deuterium is associated with the use of deuterated substances–new compounds or obtained by substitution of protium in known protium analogues. The KIE is presented, which is expressed in a decrease in the biotransformation rate as a result of deuteration, it allows predicting a rapid development of the new direction in the development of drugs. Having an identical therapeutic effect, deuterated analogs provide improved pharmacokinetic characteristics, such as reduced toxicity, blocked epimerization of optically active substances, and a change in the mechanisms of biotransformation. The obtained results make it possible to predict the mechanisms of the effect of deuterium on the biochemical transformations of pharmaceutical substances in the organism

Аэроионы и среда обитания человека

Conformity to hygienic air ion formula of working offices, classrooms and chemistry laboratory in which the work with volatile organic solvents are conducted, as well as to a city apartment and a country house is investigated. Concentration of air ions was measured using the counter «Sapphire 3М». Parameters of the air ionization: concentration of ions of both charges (N and N-, ions/cm3) and unipolarity coefficient (K = N/N-) were evaluated. The quantity of air ions did not correspond to sanitary and hygienic standards in all areas except of a country house. Additional artificial air ionization and ventilation in classrooms and chemistry laboratories offered to use.Исследовано соответствие санитарно-гигиеническим нормам аэроионного состава воздуха рабочих кабинетов, учебных аудиторий и химической лаборатории, в которой проводятся работы с летучими органическими растворителями, а также городской квартиры и загородного дома. Для измерения концентрации аэроионов использовали счетчик «Сапфир 3М». В ходе эксперимента оценивали параметры ионизации воздуха: концентрацию аэроионов обоих зарядов (N+ и N-, ионов/см3) и коэффициент униполярности (К = N+/N-). За исключением загородного дома, во всех помещениях количество аэроионов не соответствовало нормативным требованиям (СанПиН-2003). Предложено использование дополнительной искусственной ионизации воздуха и принудительной вентиляции в аудиториях и химических лабораториях

APPLICATION OF MATHEMATICAL MODELING AND PHYSICO-CHEMICAL ANALYSIS METHODS IN THE PREDICTION OF BIOLOGICAL ACTIVITY AND QUALITY CONTROL OF GOSSYPOL DERIVATIVES

Objective: The purpose of this work was to evaluate in silico biological activity profiles of real and virtual molecular structures of gossypol derivatives and to develop methods of Physico-chemical analysis to control their quality. Methods: Substance of gossypol-acetic acid (GAA) and 14 virtual derivatives; PASS and ChemicDescript QSAR methods; low angle and dynamic laser light scattering (LALLS, DLS) methods; IR Spectroscopy–Cary 630 Fourier Transform IR Spectrometer, UV spectrometry–Cary-60 spectrophotometer, Optical microscopy (Altami BIO 2 microscope); Spirotox method for a sample’s biological activity. Results: A distance-based topological Balaban index (J) was successfully selected by ChemicDescript analysis; the Pa meaning by PASS Online program showed maximum (from 0.8 to 0.9) variations of antitumor and antiandrogenic and minimum of antiviral activities of GAA derivatives (Pa<0.5) despite the existing literature data. Microscopy and DLS methods demonstrated the values of high powder dispersion d=0.8 nm and weak stability of colloidal particles =-0.9 mV. According to UV data =42.4±0.8 (100 ml·g-1·cm-1) at λmax=380 nm. The LALLS method determined the GAA dissolution rate constant in ethanol: k=0.041±0.004 s-1. The calculated activation energy values of cell biosensor death process in 1 mmol solution of GAA in N,N-DMF: °bsEa=174.36±0.45 kJ·mol-1 in comparison with the solvent medium: °bsEa=213±1.55 kJ·mol-1 Conclusion: The developed approach of chemometric, laser and biotesting methods can be used for the identification of biologically active properties, as well as for qualitative analysis within the development of the standard for the pharmaceutical substance of natural polyphenols

Влияние специфических поверхностных структур клеток Escherichia coli на процесс образования полицеллюлярных форм в жидкой питательной среде

Capacity of Escherichia coli strain bacteria containing F-like plasmids to create polycellular forms was studied. The influence of conjugative pili produced under genetic control of such plasmids on the efficiency and character of creating cell associates was shown.Изучена способность бактерий штамма Escherichia coli, содержащих F-подобные плазмиды, образовывать полицеллюлярные формы. Показано влияние конъюгативных пилей, синтезируемых под генетическим контролем таких плазмид, на эффективность и характер образующихся клеточных ассоциатов