In this work, we investigate the anti-inflammatory effect of the HLDF-6-H peptide (Thr-Gly-Glu-HseHis-Arg) in a model of the presymptomatic stage of Parkinson’s disease (PD). This peptide is a bioactive fragment of the HLDF leukocyte differentiation factor. The presymptomatic stage of PD was modelled by introducing moderate doses of MPTP toxin to C57Bl/6 mice. HLDF-6-H was administered intranasally at a dose of 300 μg/kg daily for three weeks. The severity of depression was determined by the time of immobilization in a Porsolt test. The serum levels of 10 steroids were determined using MS analysis. The activity of inflammatory markers leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) was determined using kinetic methods. The development of an inflammatory process under the influence of MPTP is evidenced by an increase in the activity of α1-PI; the anti-inflammatory effect of the peptide is manifested in a decreased activity of LE and α1-PI. At the same time, the immobilization time of mice in a Porsolt test positively correlated with α1-PI and LE, which may indicate the role of inflammation in the development of depression. A positive correlation between estradiol and LE was found in the general sample of mice. In the group of mice treated with the peptide, α1-PI correlated positively with corticosterone and negatively with estradiol and androstenedione. It can be assumed that the anti-inflammatory effect of the peptide is associated with the regulation of corticosteroids and sex hormones. Hence, the applied experimental model confirmed the development of a depressive-like syndrome and neuroendocrine disorders in the presymptomatic stage of PD. The long-term use of HLDF-6-H has antidepressant and anti-inflammatory effects.

The development of therapeutic drug monitoring of methotrexate (MT) remains an important and unresolved problem.Objective: to study the dynamics of the concentration of MT metabolites in groups of patients with different responses to MT therapy, to identify the clinical features of these groups.Patients and methods. The study included 79 patients with rheumatoid arthritis (RA), including 65 (82%) women and 14 (18%) men (mean age 53±11 years). MT monoglutamate was measured in erythrocytes (ER) and mononuclear cells (MO), as well as the main MT metabolites: polyglutamates with 2, 3 and 4 glutamate residues (MTPG2-4), as well as 7-hydroxymethotrexate (7-OH-MT) after 4, 12, 24 and 36 weeks after the start of MT treatment.Results and discussion. Among the patients who completed the 24-week follow-up, 34 responded to MT therapy (Group 1) and 36 did not respond to it (Group 2). Patients of the two groups did not differ in the concentration of various metabolites of MT after 4 weeks, age, body mass index, duration of RA, DAS28 value, radiological stage, functional class, presence of extra-articular manifestations, single and cumulative doses of MT. In the 1st group after 12 weeks of therapy, a higher concentration of 7-OH-MT (ER) was detected, after 24 weeks – a higher concentration of 7-OH-MT (MO) and a lower level of MTPH3 (ER).Conclusion. The concentration of 7-OH-MT after 12 and 24 weeks of therapy was higher in the group of patients who responded to therapy. 7-OH-MT appears to be a more persistent metabolite of MT and therefore more applicable for therapeutic drug monitoring of MT. The level of MT and its metabolites (MT monoglutamate, MTPG2 and 7-OH-MT) gradually decreases over time in responders to therapy. A 7-OH-MT concentration of 14.5 nmol/l may be a predictor of a good response to MT therapy.

Objective: to assess the time course of changes in the concentration of methotrexate (MTX) and its main metabolites in the red blood cells (RBC) and mononuclear cells (MNC) of patients with rheumatoid arthritis (RA), by taking into account individual characteristics (age, statin therapy, and smoking).Patients and methods. The investigation enrolled 33 MTX-treated patients (mean age 53.2±11.7 years) with RA, who underwent therapeutic drug monitoring to measure the RBC and MNC concentrations of free MTX and MTX polyglutamates (MTXPGs) with 2, 3, and 4 glutamate residues (MTXPG 2–4) in using tandem chromatomass spectrometry after 4, 12, and 24 weeks of therapy.Results and discussion. Following 12 weeks, the concentration of MTXPG4 in the MNC was higher in patients taking statins, while that of MTX and MTXPG2 in the RBC were significantly lower than in smokers. At 24 weeks, older patients were observed to have a higher MTX level and a lower MTXPG4 concentration in the RBC.Conclusion. After 24 weeks of therapy, the RBC concentration of MTPG4 was lower and that of MTX was higher in older patients than in others, which confirms data on a slower MTX metabolism in the elderly. The use of statins is likely to have a positive impact on the accumulation of MTXPG. There is a statistically significantly lower RBC concentration of MTXPG in at 12 weeks of therapy.

Therapeutic control of the methotrexate (MT) polyglutamates (MTPG) level in erythrocytes can be an objective marker of the effective dose of MT prescribed for rheumatoid arthritis (RA).Objective: to assess the relationship between the level of MTPG in red blood cells and efficacy of the MT dose used by RA patients.Subjects and methods. The study included 60 patients with RA (44 women and 16 men over 18 years) who met the criteria of the American College of Rheumatology and the European League Against Rheumatism (ACR/EULAR) 2010 and received MT ≥20 mg/week subcutaneously for ≥12 weeks. The patients were divided into two groups of comparable age, sex, alcohol intake, number of smokers, body mass index (BMI), depending on the presence (group 1; n=30) or absence (group 2; n=30) of the effect of MT, according to the EULAR efficacy criteria (DAS28). The concentration of MTPG (total MTPG and metabolites of MTPG 1, 2, 3, 4, 5) was determined in erythrocytes by high-performance liquid chromatography with mass spectrometric detection.Results and discussion. It was found that the levels of total MTPG and MTPG1, 2, 3, 5 in erythrocytes did not differ in groups of responders and nonresponders, and the dose of MT was comparable in both groups. At the same time, the level of MTPG4 in the first group was significantly higher (26.4±6.1 nmol/l; p=0.023) than in the second one (22.1±6.8 nmol/l). Analysis of the ROC curve showed that the values of MTPG4 <22.5 nmol/l corresponded to the absence of effect of MT. The area under the curve was 0.672 (95% confidence interval 0.536–0.808 (p=0.022), sensitivity 77%, specificity 53.3%.Conclusion. For effective treatment of patients with RA MT dose should provide MTPG4 level in red blood cells ≥22.5 nmol/l.

В статье обсуждаются данные терапевтического лекарственного мониторинга клозапина, полученные у 37 пациентов (всего 98 образцов) с различными формами шизофрении, проходящих лечение в условиях стационара. Установлен тип зависимости между концентрацией клоза пина, его метаболита норклозапина и дозой препарата. Материалы и методы. Для количественного определения клозапина и норклозапина в крови применяли метод высокоэффективной жидкост ной хроматографии с масс спектрометрическим детектированием. Мониторинг с последующим анализом данных сопровождался необходимой клинической информацией в виде специальных форм запроса – карт мониторинга. Результаты и обсуждение. Полученные данные уровня концентрации распределились следующим образом: 16 % находились в терапевтиче ском, 51 % – в субтерапевтическом (< 350 нг/мл) и 29 % – в условно токсическом диапазоне (> 600 нг/мл), частота случаев предполагаемой некомплаентности не превышала 3 % при назначении клозапина в дозах от 25 до 350 мг/сут. Средние значения концентрации клозапина и норклозапина были выше у некурящих пациентов по сравнению с курильщиками. Выводы. Распределение по концентрациям для клозапина и норклозапина имеет в основном нормальный характер с небольшой асимметрией в сторону высоких значений. Терапевтический лекарственный мониторинг является необходимым инструментом для индивидуализации и пер сонализации фармакотерапии при лечении шизофрении.

The article discusses therapeutic drug monitoring (TDM) data for clozapine obtained in patients with various forms of schizophrenia undergoing treatment in hospital. Clozapine and its metabolite norclozapine concentrations and their response to the administered drug doses have been studied. Clozapine and norclozapine assay in human blood has been performed by high performance liquid chromatography with mass spectrometric detection. Monitoring and subsequent data analysis has been supported with necessary clinical information in the form of special requests - TDM cards. The obtained data on concentration levels have been arranged as follows: 38.64 % within therapeutic range, 38.64 % within subtherapeutic range (<350 ng/ml) and 22.73 % within arbitrary toxic range (600 ng/ml) when administering 25 to 30 mg of clozapine daily. Average concentrations of clozapine and norclozapine were higher in non-smokers compared to smokers.

General analytical trends and approaches to quantitative determination of atypical neuroleptics (antipsychotics) in biological matrices were presented. Various analytical methods and sample preparation procedures for quantitative chemical analysis of these compounds were described.