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Introduction. "Eltrombopag" is a thrombopoietin receptor agonist (TPO-RA) that is approved for the treatment of immune thrombocytopenic purpura (ITP). According to the literature, very few analytical methods for determining eltrombopag in biological samples have been reported. To study the pharmacokinetics of new formulations of eltrombopag, a sensitive and specific method is required that allows one to accurately determine the concentration of eltrombopag in human blood plasma. Normally, HPLC methods should provide time, accuracy, and sensitivity as a result, it is necessary to develop fast or ultra-fast methods such as LC-MS/MS without any loss in sensitivity or separation efficiency.Aim. We aimed to develop and validate a method for the quantitative determination of eltrombopag levels in human plasma by using HPLC with mass spectrometric detection for performing the analytical part of pharmacokinetic studies.Materials and methods. Eltrombopag levels were determined in human plasma by HPLC with mass spectrometric detection. The samples were prepared using protein deposition.Results and discussion. The method was validated for selectivity, matrix effect, calibration curve, accuracy, precision, the limit of quantification, carry-over effect, and sample stability.Conclusion. The method for the determination of eltrombopag levels in human plasma has been developed and validated by HPLC-MS. The analytical range of eltrombopag levels in human plasma was 10–6750 ng/ml. This method could be used to determine eltrombopag levels in plasma for PK and BE studies.

Introduction. Non-neurogenic lower urinary tract symptoms (LUTS) are a serious polyetiologic problem in the male population. The side effects of the medication agents used to treat LUTS significantly reduce treatment compliance. According to the literature data, the frequency of refusal for the proposed treatment during the year varies from 20 to 80%. Several studies have shown the benefits of herbal medicine for LUTS concerning the fewer side effects and increased adherence to treatment. However, to obtain a high-level recommendation base, clinical trials are required.Purpose of the study. To evaluate the effectiveness of Gardaprost® in LUTS combination therapy.Materials and methods. The study included 57 men aged 60 – 70 years with diagnosed medium- or large-volume benign prostatic hyperplasia (BPH) and moderate-to-severe LUTS according to I-PSS, morphologically excluded prostate cancer, without urinary infection signs. The patients were randomized into two follow-up groups. Tamsulosin 0.4 mg q.d. was prescribed to patients in the control group. Patients of the main group received Gardaprost® 0.4 mg q.d. in addition to Tamsulosin. The follow-up period in both groups was one-year. The statistical analysis includes data from 56 men. To evaluate therapy at the screening visit and on days 180 and 360, I-PSS, urination diary, IIEF-5, urinalysis, prostate-specific antigen, uroflowmetry, ultrasound were analyzed. Paired t-test and one-way ANOVA test were used to determine intergroup differences in normally distributed variables. For variables with a distribution other than normal, Friedman's two-way ANOVA for related samples was used. Events with a probability greater than 95% were considered statistically significant.Results. At the time of inclusion in the study, the groups were comparable concerning the control parameters. In the main group, there was a more pronounced positive dynamics in the I-PSS score, maximum urine flow rate, and post-void residual urine volume, which corresponded to 7.9 ± 2.1 points, 18.0 ± 7.3 ml/sec, 23.6 ± 13.6 ml vs 19.7 ± 7.2 points, 10 ± 3.5 ml/sec, 65.9 ± 33.2 ml in the main and control groups, respectively (p

The correction of free radical oxidation processes is one of the most promising strategies of neuroprotection in acute cerebrovascular disorders.The aim of the study is an experimental study of the neuroprotective effects of 3-hydroxypyridine and erythropoietin derivatives, as well as their combined use.Materials and methods. The study was performed on 109 male Wistar rats. The neuroprotective effect of the substances was studied on a hemorrhagic stroke model. The study drugs were administered to the animals intraperitoneally. Carbamylated darbepoetin was administered three times in advance at the dose of 100 µg/kg within intervals of 3 days, the last injection took place 1 hour before the operation (the total dose was 300 mg/kg). Etoxidol was administered once 1 hour before the surgery at the dose of 50 mg/kg. The survival rate, behavioral features and the state of the animals on the 1st, 3rd, 7th and 14th days were recorded, and the morphological assessment of the brain was carried out.Results. The investigated substances had a positive effect on both the survival rate of the animals during the first day and on the 14th day. The best survival rates on the 14th day were recorded in the group of a combined use of ethoxydol and carbamylated darbepoetin (75%). Thus, in this group of rats, a faster recovery of neurological disorders was already distinguished from the first day on. By the 7th day, more than 50% of the rats receiving the combination of the studied drugs, had had a slight neurological deficit (up to 3 points on the McGrow scale); by the 14th day there had been only minor changes in the neurological status in the rats of this group. A pronounced neuroprotective effect of the combination of 3-hydroxypyridine and erythropoietin derivatives has been confirmed by a histological examination of brain slices – a more rapid decrease in the size of perifocal edema and microcirculation disorders, less damage to neurons and glial elements, and faster processes of resorption and organization of hemorrhage. A macroscopic examination of the brain sections stained with triphenyltetrazolium chloride of the dying rats, showed that perifocal necrosis had been the main cause of high mortality in the control group after the 3rd day.Conclusion. As a result of the experiment, the nephroprotective effect of the studied derivatives of 3-hydroxypyridine and erythropoietin has been proved. Moreover, the combination of these drugs has shown a greater neuroprotective activity than their isolated use. The additive effect of these drugs was due to their action mechanism resulting from the synergism of various structures and components of the cells.

The aim of the study was to test whether P-αB can be positioned as a preventing and treating agent for cardiovascular diseases.Materials and methods. The study was performed on sexually mature male Wistar rats. Endothelial dysfunction was modulated by a 7-days intraperitoneal administration of L-NAME at the dose of 2.5 mg/100 g. P-αB, or erythropoietin (EPO), was used for therapy at the dose of 2.5 µg/100 g × 3 times for 7 days, the total dose was 7.5 µg/100 g. The function of endothelium was estimated by an endothelium-dependent and endothelium-independent vasodilation. In addition, a histological assessment of the abdominal aortic wall state and the analysis of eNos, Tnf and Il-1β genes expression were performed. To estimate prothrombotic properties, P-αB and EPO were administered, at the doses of 2.5 and 5 µg/100 g (3 times a day for 7 days, the total doses were 7.5 µg/100 g and 15 µg/100 g, respectively) and on the 8th day, the time of ferric (III) chloride-induced carotid artery thrombosis was estimated.Results. Theresults of the functional tests for endothelium-dependent and endothelium-independent vasodilatation, as well as the histological picture of the aorta have evidenced that P-αB and EPO do not affect L-NAME-induced hypertension but improve the endothelium function. At the same time, P-αB shows a significantly higher endothelial-protective activity, reducing the coefficient of endothelial dysfunction from 5.1±0.15 to 2.72±0.12. In addition, P-αB has significantly increased the expression of eNos and reduced the expression level of Tnf and Il-1β mRNA genes. Carrying out Ferric (III) chloride-induced carotid artery thrombosis has revealed that P-αB (5 µg/100 g × 3 times a day for 7 days, total dose was 15 µg/100 g) has a lower but statistically significant prothrombotic activity than EPO.Conclusion. P-αB can be positioned as an atheroprotector because of its ability to prevent the death of endothelial cells, as well as to reduce remodeling and proinflammatory activation of the vascular wall. However, the prothrombotic properties of P-αB limit its use as a preventing and treating agent for atherosclerosis-associated diseases.

The aim of the article is to evaluate Bischofit gel reparative activity in a linear wound model in rats.Materials and Methods. The study was conducted on 36 male Wistar rats weighing from 193 to 218 grams. On the 8th day after modeling a linear wound defect 50±1 mm long, the reparative effect of bischofite, Actovegin and Contractubex in the gel compositions was evaluated. The evaluation was carried out using: the following methods: 1) studying the physicomechanical characteristics of the wound defect (a wound-tearing machine Metrotest REM-0.2-1); 2) morphological examination of the skin graft taken from the wound area (stained with hematoxylin-eosin and Van Gieson‘s solution); 3) determining the ratio of collagen types I and III in a polarizing microscope (the picrosirius was red); 4) cоlorimetric analysis of the hydroxyproline concentration in the wound surface tissues.Results. On the 8th day, the wound defects sampled from the bischofite treated animals, were characterized by the most pronounced strength (the average force at the rupture moment was 13.70 N), which was significantly higher (p Actovegin (0.67±0.017) > Control (0.56±0.012) > Contractubex (0.38 ±0.020).Conclusion. The carried out study showed that Bischofit has a pronounced ability to stimulate the regeneration of the skin wound defect. Hereby, the reference drug Actovegin showed less activity, and Contractubex worsened wound healing.

This article contains the results of research of the endothelial dysfunction arising during the modeling of thermal local asphyxia of kidney and possibilities of their correction by distant ischemic and pharmacological preconditioning. The modeling of thermal local asphyxia of kidney is characterized by the disturbance of microcirculation and expression of eNOS in the kidney tissue. The usage of distant ischemic preconditioning and phosphodiesterase inhibitors type 5 sildenafil and tadalafil leads to pronounced correction of microcirculation dysfunction and activity of eNOS. During the modeling of thermal local asphyxia of kidney against endothelium dysfunction caused by ADMA-like L-NAME induced deficiency of nitric oxide more pronounced dysfunction of microcirculation and activity of eNOS are observed. The usage of distant ischemic and pharmacological preconditioning with the help of phosphodiesterase inhibitors type 5 in this type of pathology led to pronounced correction of microcirculation dysfunction and activity of eNOS. The injection of glibenclamide blocker of ATP – dependent K+ channels during the correction of the disturbance caused by the modeling of thermal local asphyxia of kidney with the help of distant ischemic and pharmacological preconditioning leads to the decrease of its efficiency.Disclosure: The study did not have sponsorship. The authors have declared no conflicts of interest.

Many recent studies have confirmed the effect of NO and that its relative signaling pathway is important for preconditioning of the cardioprotective effect. Mitochondria as a target for the cardioprotective effects of nitric oxide in ischemia-reperfusion injury is considered to be the ultimate goal of cardioprotection. During preconditioning, signaling is initiated from the sarcolemmal membrane and then spread into the cytoplasm via many series of enzymes, including nitric oxide synthase (NOS), soluble guanylyl cyclase (sGC), and protein kinase G (PKG). As a result, the signal is transmitted into the mitochondria, where the cardioprotective effect occurs. It is now well established that mitochondria act to protect the heart against ischemia-reperfusion injury via the opening of the mitochondrial ATP-sensitive K+ channel and the inhibition of mitochondrial permeability transition. This knowledge may be useful in developing novel strategies for clinical cardioprotection from ischemia-reperfusion injury.

The possibility of vardenafil using for pharmacological preconditioning in surgery is studied. Experiment was carried out on Wistar rats. The influence of vardenafil on the survival of pedicled isolated skin flap and the state of the microvasculature in the ischemic skeletal muscle was defined in comparison with the effect of distant ischemic preconditioning. It was revealed that vardenafil improves survival of pedicled isolated skin flap and stimulates angiogenesis in the ischemic leg muscles of rat.

The study demonstrated that eight weeks after ovariectomy in female white Wistar rats endothelial dysfunction of bone microvasculature and deterioration of regional blood flow in bone develops, leading to the emergence of generalized osteoporosis. Nanoparticulated forms of losartan and resveratrol, possessing endothelioprotective action, effectively prevents the reduction of regional microcirculation in bone tissue, keeping it at the level of intact rats. It is allowed to maintain an adequate level of bone remodeling processes, which manifested in slowing of the thinning of bone trabeculae and preventing the occurrence of their microfractures.