- Comparative study of pharmacokinetics
According to the Manual for conducting preclinical studies of drugs (Part one. M.: publishing house “Grif and K”, 2012, P. 854), comparative study of pharmacokinetics means the research of the drug in a single dose using one animal species accompanied by an analysis of one type of biomaterial – blood plasma or serum. This study includes:
- a design of the quantitative determination method of an active substance in bioliquid (plasma or serum), pilot experiment to determine the dose for administration;
- a study of pharmacokinetics of a single dose administration, animals used are rabbits, rats or mice, blood sampling is not less than 10 time points;
- an analysis of data obtained;
- a calculation of pharmacokinetic parameters by the model-independent method of statistical moments, writing the report based on the experimental data and in accordance with GOST 7.32-2001 “Report on research work”
Study on pharmacokinetics of the original drug
The study is carried out according to the Manual for conducting preclinical studies of drugs (Part one. M.: publishing house “Grif and K”, 2012, P. 845—855). The study includes the following scope of work:
- design and validation of the quantitative determination method of an active substance in blood plasma (the main type of biomaterial);
- pilot experiment aimed at clarifying the drug doses for further analysis of pharmacokinetics;
- adaptation and revalidation of the quantitative determination method of an active ingredient in other types of biological material;
- assessment of linearity of drug pharmacokinetics provided a single dose administration using three dose levels. Distribution of active substance in the tissues (brain, liver, kidneys, spleen, skeletal muscles, heart) is studied in parallel;
- study of the drug elimination provided a single dose administration (analysis of one type of excreta – urine);
- repeated administration of the drug at a single dose (in case of an approved linearity of pharmacokinetics) or using two levels of doses (it is necessary, if pharmacokinetics is nonlinear). Analysis of one type of biological material – blood plasma;
- study of pharmacokinetics and relative/absolute bioavailability of test chemical provided a single dose administration;
- calculation of pharmacokinetic parameters by a model-independent method of statistical moments, assessment of pharmacokinetics linearity, writing a report based on experimental data and in accordance with GOST 7.32-2001 “Report on research work”
To perform the work we have qualified and trained experts and the appropriate equipment for quantitative determination of drugs and their metabolites in vivo and in vitro using HPLC, IFA, HPTLC, capillary electrophoresis methods.
- Карлина М.В., Пожарицкая О.Н., Косман В.М., Макарова М.Н., Шиков А.Н., Макаров В.Г., Забозлаев А.А. Экспериментальная фармакокинетика препарата “Валеокор-Q10”. Фармация. – 2012.- N- С. 35-37.
- Pozharitskaya O.N., Kosman V.M., Karlina M.N., Shikov A. N., Makarov V.G., Djachuk G.I.. Method development and validation of an HPLC assay for the detection of hopantenic acid in human plasma and its application to a pharmacokinetic study on volunteers.// Acta Chromatographica. – 2011. – Vol.23, Is.3. – P. 403-414.
- Карлина М.В., Пожарицкая О.Н., Косман В.М., Шиков А.Н., Лазукина М.А., Дьячук Г.И. Исследование фармакокинетики гопантеновой кислоты при введении внутрь // Экспериментальная и клиническая фармакология. – 2010. – Т. 73, №8. – С. 46-48.
- Pozharitskaya O.N., Karlina M.V., Shikov A.N., Kosman V.M., Makarova M.N., Makarov V.G. Determination and pharmacokinetic study of taxifolin in rabbit plasma by high-performance liquid chromatography// Phytomedicine.- – Vol. 16 (2/3).- P. 244-251.
- Karlina M.V., Pozharitskaya O.N., Shikov A.N., Kosman V.M., Makarova M.N., Makarov V.G. LC method for quantification of lutein in rat plasma: validation and its application to pharmacokinetic study// Chromatographia. – 2008. – Vol. 68 (11/12). – P. 949-954.
- Карлина М.В., Пожарицкая О.Н., Дадали Ю.В., Косман В.М., Шиков А.Н., Макаров В.Г. Светопоглощающие и антирадикальные свойства продукта на основе лютеина и зеаксантина в опытах in vitro и оценка кинетики каротиноидов при однократном пероральном применении у крыс// Вопросы питания. – 2008. – Т. 77, N 3. – С. 34-38.
- Pozharitskaya O.N., Karlina M.V., Shikov A.N., Kosman V.M., Makarova M.N., Makarov V.G. Determination of icariin in rat plasma by reverse-phase high-performance liquid chromatography after oral administration of a lipid-based suspension of Epimedium koreanum extract// Biomedical Chromatgraphy. – 2008, Vol. 22(6).– P. 625-629.
- Pozharitskaya O., Karlina M., Ivanova S., Shikov A., Makarov V., Tikhonov V. Plasma analysis of boswellic acids by HPTLC: a useful technique for pharmacokinetic studies./ Abstracts book of the 5th International Symposium on Chromatography of Natural Products (ISCNP), Lublin (Poland), 19-22 June 2006.- P.183.
- Карлина М.В., Пожарицкая О.Н., Косман В.М., Шиков А.Н., Забозлаев А.А., Макарова М.Н., Макаров В.Г. Модель in vitro для оценки скорости растворения гидрофобных веществ из таблеток для рассасывания (lozenges) на примере коэнзима Q10, in vivo/in vitro корреляция // Химико-фармацевтический журнал. -2012. –Т. 46, № 7. –С. 52-55.
- Карлина М.В., Пожарицкая О.Н., Косман В.М., Макарова М.Н., Шиков А.Н., Макаров В.Г., Забозлаев А.А. Экспериментальная фармакокинетика препарата «Валеокор Q10» // Фармация. -2012, № 8. –С. 35-37.
- Карлина М.В., Косман В.М., Пожарицкая О.Н., Шиков А.Н., Макарова М.Н., Макаров В.Г., Балабаньян В.Ю. Экспериментальное исследование фармакокинетики рифабутина в липосомальной форме // Фармакокинетика и фармакодинамика. – 2013, №1(6). – С. 37-41.
- Kosman V.M., Faustova N.M., Karlina M.V., Pozharitskaya O.N. Immunoassay as analytical method in pharmacokinetic and toxicokinetic invastigations // Abstracts. Phitopharm. 2016. Obzory po klinicheskoj farmacologii i lekarstvennoj terapii. – 2016; 14. – P. 28.IMMUNOASSAY AS ANALYTICAL METHOD IN PHARMACOKINETIC AND TOXICOKINETIC INVESTIGATIONS
- Kosman V.M., Faustova N.M., Urakova I.N., Karlina M.N., Makarov V.G. Dypeptydylpeptidase IV activity ingibition after oral administration to rabbits of Strongylocentrotus droebahiensis gonads extract as possible biomarker of pharmakokinetics. Razrabotka i registratsiya lekarstvennykh sredstv = Drug development & registration. – 2020; 9(3). –P. 158-165. https://doi.org/10.33380/2305-2066-2020-9-3-158-165 Abstract. Introduction. The pharmacokinetic profiling of active compounds is necessary for drug development and application. Comlex extract of biologically active compounds was isolated from gonads of green sea urchin Strongylocentrotus droebachiensis from Barents Sea. It contains fatty acids, carotenoids and tocopherols and have inhibition activity to ensyme dypeptydylpeptidase IV (DPP-4). Traditional approaches to pharmacokinetic study based on chromatographic methods are not effective for such complex mixtures because they had not enough selectivity and sensitivity. Methods of immunoassay for special enzyme, bioassay, etc. may be used as an alternative way. Aim. The aim: to find approach to pharmacokinetic study of sea urchin gonads extract in rabbits after a three doses oral administration. Materials and methods. Various spectroscopic and chromatographic (HPLC and TLC) methods were used for chemical characteristic of sea urchin gonads extract ant different target analytes quantification in biosamples. The pharmacokinetic of sea urchin gonads extract was studied after single dose oral administration to male rabbits. The correlation between concentration of sea urchin gonads extract in the blood plasma and its biological activity marked by DPP-4 activity was established. The activity of DPP-4 was determined by the chromogenic optical method. Results and discussion. Main groups of sea urchin gonads extract chemical compounds were characterized. Total peptides content was 15–22 %; α-tocopherol content – 0.05–0.15 %, total tocopherols content – 0.23–0.38 %; total carotenoid content – 0.005–0.07 %; total fatty acids content calculated to linolenic acid – 11,03 to 12,74 %. After sea urchin gonads extract chemical composition results it was established that concentrations of identified individual biologically active compound are low, there is no dominant compound or group of compounds responsible for pharmacological activity of sea urchin gonads extract which may be unambiguously chosen as target analyte for its pharmacokinetic study. Approach based on correlation of special marker activity (DPP-4) and extract concentration appears to be the best for pharmacokinetic study of complex extract from green sea urchin gonads. The method for the quantitative determination of sea urchin gonads extract in the blood plasma of rabbits by its effect on DPP-4 activity was developed. The method was validated by parameters: selectivity, lower limit of quantification, linearity range, accuracy, and precision. The pharmacokinetics was linear in the dose range of 5–25 mg/kg after oral administration. The mean maximal concentration in plasma (Cmax) was dose dependent and it was 37.12–114.71 µg/mL for various doses, area under the curve (AUC0-24) was 192.92–597.14 hµg/mL, mean time to reach maximum plasma concentration (Tmax) was 3–3.5 h, half-life (T1/2) was 7.88–9.89 h, and the mean retention time (MRT) was 1.73–14.31. Conclusion. After nontraditional approach based on correlation of special marker activity (DPP-4) and active substance concentration the plasma pharmacokinetics of sea urchin gonads extract after oral administration to the rabbits in three doses was characterized. Similar approaches may be effective for compounds and complex mixtures when it is difficult or impossible to analyze them traditionally by chromatographic (HPLC-UV/FL/MS, GC-MS, etc.) methods [Full text is available in Russian].
- Kosman V.M., Tyutina K.V., Dadali Yu.V., Karlina M.V. Pronitsaemost’ aglikonov nekotorykh flavonoidov v Saco2-modeli «Novye dostizheniya v khimii i khimicheskoi tekhnologii rastitel’nogo syr’ya»: materialy VIII Vserossiiskoi konferentsii. Barnaul. 5-9 October 2020.
- Kosman V.M., Karlina M.V. Residual protein content in the blood plasma biosamples of laboratory animals (rabbits) after their preparation for HPLC-UV analysis. Problems of biological, medical and pharmaceutical chemistry. – 2020; 23(10). –P. 53-58. https://doi.org/10.29296/25877313-2020-10-08 Abstract. Relevance. The main limitation of the sensitivity of HPLC-UV techniques widely used for the analysis of drugs in biological samples (blood plasma, tissues and organs) is due to the significant background influence of biologic matrices. Precipitation of proteins is the simplest, most versatile and reasonably efficient way to prepare biosamples for HPLC analysis. Information on the effectiveness of the protein deposition methods used and the level of their residual content in the analyzed biosamples is limited in the available literature. The aim: to evaluate the residual protein content in the blood plasma biosamples of laboratory animals (rabbits) after samples preparation for HPLC-UV analysis. Material and methods. The residual protein content in the rabbits blood plasma biosamples was evaluated by spectroscopic methods after samples treatment for HPLC-UV analysis, including precipitation with acidic solutions, acetonitrile and methanol. Results. The Bredford method has been shown to produce lower level results compared to the direct spectrophotometry method. Using different precipitation agents, their ratios and two centrifugation modes, the residual protein content determined by the Bradford method was about 0.02–0.4 mg/ml, indicating almost complete precipitation of the proteins (more than 99.5%) and confirming the correctness of using the precipitation sample preparation for further HPLC-UV analysis of the samples. Most preferred in terms of minimum residual protein content is the use of acetonitrile as a precipitation agent. The highest level of protein was found in the samples after treatment with 15% chloric acid, i.e. the use of this precipitation agent is least desirable for further HPLC analysis. Conclusions.Precipitation is effective way for sample protein removing; established features of various precipitation agent application may be used for development of bioanalytical methods, based on HPLC-analysis [Full text is available in Russian].