The results of studying magnetic resonance imaging (MRI) of rat liver performed with the original hepatotropic paramagnetic contrast agent (PMCA) Mn-DTPA-GDOF, the Mn(II) complex of 2-[2-carboxymethyl-(4-hexadecyloxyphenylcarbamoylmethyl)-aminoethyl]-aminoethyl-(4-hexadecyloxyphenylcarbamoylmethyl)aminoacetic acid, are presented. Mn-DTPA-GDOF was used as a colloidal solution (0.025 M, pH 8 – 9) with a free Mn(II) content <0.03

РЕЗЮМЕАктуальность.Нарушение метаболических процессов печени в результате воздействия алкогольной интоксикации снижает когнитивное функционирование и индуцирует когнитивные расстройства.С выраженностью метаболических расстройств связана тяжесть осложнений алкоголизма.У больных алкоголизмом с коморбидным поражением печени признаки когнитивных нарушений наблюдаются уже на ранних стадиях заболевания и сопровождаются расстройствами внимания, памяти, восприятия, что в дальнейшем зачастую приводит к социальной дезадаптации и инвалидизации.Цель.Выявление взаимосвязи показателей нарушения детоксицирующей функции печени и когнитивных расстройств у больных с алкогольной зависимостью для разработки алгоритма персонализированной терапии.Материалы и методы.На базе отделения аддиктивных состояний НИИ психического здоровья сформирована исследовательская выборка (n=40) из лиц мужского пола, средний возраст которых составил 42,4±11,9 года, с диагностированными по МКБ-10 психическими расстройствами и расстройствами поведения, связанными с употреблением алкоголя (F10), нарушениями детоксицирующей функции печени и когнитивными расстройствами.Для оценки динамики психического состояния у пациентов с алкогольной зависимостью дважды (при поступлении и после комплексной терапии) проведено психологическое тестирование с использованием шкалы тревоги Гамильтона.Скрининг когнитивных нарушений осуществляли по Монреальской когнитивной шкале (MoCA), оценку кратковременной и долговременной памяти -по психодиагностическим тестам (Пиктограммы, Заучивание 10 слов).Обсессивно-компульсивная шкала (ОКШ) влечения к алкоголю использовалась для самооценки идеаторных проявлений отношения к алкоголю.Исследовались в динамике биохимические показатели крови (АЛТ, АСТ, общий билирубин, прямой и непрямой билирубин, холестерин, щелочная фосфатаза

The search for new drugs to reduce the toxic effects of alcohol on the body is based on the creation of various pharmaceutical agents that affect the toxicokinetics and/or toxicodynamics of ethanol. A differentiated approach is needed in the pharmacotherapy of alcoholism due to the different clinical typologies of the condition. Compared with synthetic drugs, herbal remedies have multiple types of activity, which complement their efficacy. Research and development of technology for the manufacture of new drugs based on the hairy agrimony ( Agrimonia pilosa ) are promising. Experimental studies in which white outbred mice were tested in an open field showed that a hairy agrimony extract reduced orientational-exploratory behavioral activity and contributed to adaptation of the animals to stress. Mice treated with agrimony extract showed a reduction in sensitivity to administration of ethanol: the latent period of falling asleep was reduced by a factor of 1.6 and the duration of sleep decreased by a factor of 3 compared with the group of mice given starch paste. Rats treated with agrimony extract at doses of 100 and 150 mg/kg showed reductions in the duration of ethanol-induced sleep by factors of 1.8 – 2.4 as compared with animals treated with St. John’s wort extract, indicating that the extract had a detoxifying effect. Hairy agrimony extract significantly reduced grooming in all groups of animals, this being associated with its anti-anxiety effect and of great importance in terms of reducing the development of delirium in acute alcohol intoxication.

Поиск новых средств для снижения токсических эффектов алкоголя на организм основан на создании различных фармацевтических средств, влияющих на токсикокинетику и/или токсикодинамику этанола. Необходим дифференцированный подход при фармакотерапии алкоголизма из-за различной клинической типологии заболевания. В сравнении с синтетическими препаратами, растительные средства обладают несколькими видами активности, дополняющими эффективность их действия. Исследование и разработка технологии изготовления новых лекарственных средств на основе репешка волосистого (Agrimonia pilosa) являются перспективными. В ходе проведения эксперимента при тестировании белых беспородных мышей в «открытом поле» установлено, что экстракт репешка волосистого уменьшал ориентировочно-исследовательскую активность поведения и способствовал адаптации животных к стрессу. У мышей, получавших экстракт репешка волосистого, снижалась чувствительность к введению этанола: латентный период засыпания сократился в 1,6 раза, а длительность сна — в 3 раза по сравнению с группой мышей, получавшей крахмальную слизь. У крыс, получавших экстракт репешка волосистого в дозах 100 и 150 мг/кг, длительность этанолового сна уменьшалась в 1,8 – 2,4 раза по сравнению с животными, получавшими экстракт зверобоя, что свидетельствует о детоксикационном эффекте данного экстракта. Экстракт репешка волосистого значительно снижал груминг во всех группах животных, что связано с его противотревожным действием и является крайне важным для редукции развития делирия при острой алкогольной интоксикации.

Molecular docking studies (in Schrödinger and Glide software) showed that the molecule m-Cl-BHU ( meta -chlorobenzhydrylurea) is complementary to the benzodiazepine binding site of GABA A receptors. Binding energy was low (-11.14 kcal/mol); m -Cl-BHU interacts with the key amino acids at the α1γ2 interface: Tyr159, Tyr209, and H101 Phe77 with high fit to the dG insertion model: 0.741. Binding of [ 3 H]flunitrazepam with the benzodiazepine site of brain GABA A receptors increased in rats with experimental alcoholism treated with m -Cl-BHU at a dose of 100 mg/kg for 14 days. Changes in pharmacokinetic parameters ( T 1/2 , Cl t , MRT , MET , and AUC ) of the model substrate antipyrine in saliva were seen in healthy volunteers and male patients with alcoholism using Galodif ( m -Cl-BHU) at a dose of 300 mg/day for 21 days. Elimination of antipyrine in patients with alcoholism was increased due to activation of microsomal cytochrome P450 in the liver oxidase system.

С помощью молекулярного докинга (программа Schrödinger, Glide) показано, что молекула м-Cl-БГМ (мета-хлорбензгидрилмочевина) комплементарна сайту связывания бензодиазепина ГАМКА рецепторов. Энергия связывания низкая (– 11,14 ккал/моль); м-Cl-БГМ взаимодействует с ключевыми аминокислотами на α1γ2 интерфейсе: Tyr159, Tyr209, H101 Phe77 с высокой степенью соответствия модели — dG вставки: 0,741. Связывание [3H]флунитразепама с сайтом бензодиазепина ГАМКА рецепторов головного мозга крыс при экспериментальном алкоголизме, получавших 14 дней м-Cl-БГМ в дозе 100 мг/кг в сутки, увеличивалось. Выявлены изменения фармакокинетических параметров (T1/2, Clt, MRT, МЕТ, AUC) модельного субстрата — антипирина в слюне здоровых добровольцев и больных алкоголизмом мужчин при применении галодифа (м-Cl-БГМ) в дозе 300 мг/сут в течение 21 дня. Элиминация антипирина у больных алкоголизмом была повышена вследствие активации микросомальной цитохром-P450 оксидазной системы печени.

Metabolites of the anticonvulsant m-CL-BHU in the process of detoxification on cytochrome P-450 are conjugated with endogenous macromolecules and form a complete stimulus for the immune system. Specific activation occurs at the time of the decrease in the activity of liver microsomal monooxygenases. This protection function is valuable in maintaining “immuno-chemical homeostasis” in the body.

Sensitization of cytochrome P-450 system to action of alcohol can become a significant problem of psychopharmacotherapy. M-chlor-benzhydrylurea – Galodif® is an efficient anticonvulsant. We investigated effect of Galodif on activity of the liver cytochrome P450 system of alcoholics from two different ethnic groups. As a test-drug antipirine was used. 68 patients (from Russian and Tatar ethnic groups) were examined. The concentration of test-drug antipirine in saliva was determined by spectrophotometry assay. Pharmacokinetic parameters were counted by model-independent method of statistical moments by K. Yamaoka: period of half-elimination (T1/2, h), total clearance (Cl t, ml/min), middle time of residual drug in organism (MRT, h), middle time of elimination (MET, h), area under the pharmacokinetic curve (AUC, mkgh/ml). Clinical monitoring provides a possibility to considerably optimize the process of treatment of alcoholic patients. We observed, that T1/2 of drug kinetic was 8,81±5,23 before treatment and 4,37±2,31* after treatment with Galodif; Clt: 113,42±38,67 and 137,37±54,00; MRT: 11,44±5,43 and 3,69±0,60* (p<0.05); MET: 6,03±2,10 and 4,64±1,83* (p<0.05); AUC: 7,05±5,74 and 6,39±2,18, respectively. Galodif causes reduction of period of half-elimination, significant decrease of middle time of residual drug in organism and middle elimination time. Drug pharmacokinetics parameters in alcoholic patients from Tatar ethnic group were as follows: T1/2: 11,19±2,95 and 2,57±0,69*; Clt: 71,108±11,58 and 116,23±9,40*; MRT: 8,66±1,13 and 2,60±0,46*; MET (h) 5,71±0,57 and 3,68±0,49*; AUC: 11,58±1,71 and 7,30±1,04*, respectively. These data suggest that the individual sensitivity of organism to the drug is caused not only by biochemical, but also by anthropo-morpho-physiological polymorphism.

Studying the effects of drugs on neuronal GABAA/benzodiazepine receptors can be the basis to develop new approaches to the treatment of this disease. Wistar rats (n = 250) used in the experimental model of alcoholism. Properties of BDR 'synaptosomal" and 'mitochondrial" types were examined in respective membrane fractions obtained from brain cortex of rats with experimental alcoholism and treating of anticonvulsant meta-chloro-benzhydrylurea (m-chBHU) by radioreceptor assay with [3H]flunitrazepam and [3H]Ro5-4864. As a result of screening in terms of consumption of 15% alcohol and water in the rat were divided into 3 groups of animals. 1st – rats preferred to ethanol in testing – 'heavy drink" rats (15% alcohol as the sole source of drinking for 10 months); 2nd – rats preferred to ethanol – 'non-heavy drink" male contained no access to the entire period of ethanol; 3rd – 'non-prefer" alcohol rats – contained in the water. Introduction of m-chBHU rats 100 mg/kg for 14 days caused a significant decrease in alcohol consumption in animals preferring alcohol (1st and 2nd groups). Comparative study of kinetic parameters of selective ligands with bran membranes showed that properties of BDR in membranes from brain cortex of male rats with different preference to alcohol and showed that affinity of BDRs was decreased, but capacity of receptors was increased in brain cortex of 'heavy drink" and 'non-heavy drink" male rats compared with 'non-prefer" alcohol rats. Administration of m-chBHU induced mediation of GABA in brain of these rats and reduced alcohol consumptions.

Chronic alcohol intake induces neuroadaptive changes in benzodiazepine receptors modulating GABA A receptors that promote alcohol addiction. Analysis of benzodiazepine receptors in the brain of Wistar rats differing by alcohol preference has demonstrated that affinity of [ 3 H]flunitrazepam and [ 3 H]Ro5-4864 binding with membrane fraction was reduced, while the density of specific binding sites in the brain cortex of heavy drinking and low drinking rats was increased in comparison with rats nonpreferring alcohol. Administration of anticonvulsant meta-chlorobenzhydryl urea increased affinity of benzodiazepine receptors in the brain cortex of heavy drinking rats, which improved GABA neurotransmission in the brain of these animals and reduced alcohol consumption.

The objectives of this study were to determine residual and absorbed levels of medroxyprogesterone acetate (MAP) after treatment with intravaginal sponges impregnated with different doses of MAP in an ewe herd and to determine the effects of such treatments upon estrus incidence, interval to estrus onset and pregnancy rate. Polyurethane sponges impregnated with 40, 50 and 60 mg of MAP were prepared. Real amounts of progestagen on sponges were checked prior to sponge treatment insertion. During autumn 1999, 608 cyclic Merino ewes were treated with intravaginal sponges impregnated with different doses of MAP (groups I, 40; II, 50 and III, 60 mg). After 14 days, sponges were removed and residual levels of MAP (RMAP) on removed sponges were measured by spectrophotometry. Real amounts for unused sponges were 39.60±1.10, 44.10±1.06 and 59.10±1.26 mg for the intended doses of 40, 50 and 60 mg MAP respectively. RMAPs were different (p<0.05) among groups (I: 17.98±1.97 mg; II: 24.32±2.03 mg; III: 34.25±3.23 mg). Absorbed levels of MAP were not different among groups (I: 21.62±1.97 mg; II: 19.78±2.03 mg; III: 24.85±3.23 mg). Artificial insemination was performed in 16 ewes from Group I, 17 ewes from Group II and 11 ewes from Group III, 12 h after estrus onset. There were no differences among groups, neither for estrus incidence (I: 79.27%; II: 77.42%; III: 80.87%) nor interval to estrus onset (I: 55.94±1.87 h; II: 56.74±1.13 h; III: 57.70±1.02 h). There were also no differences among groups for pregnancy rate (I: 43.75%; II: 52.94%; III: 45.45%). It was concluded that under similar conditions, a dose as low as 40 mg MAP could be effectively used for estrus synchronization in cyclic Merino ewes.

Cumazid, a new prophylactic immunotherapy preparation, has been created based on the total glycoside fraction isolated from Cucumaria japonica sea cucumber species. The acute and chronic toxicity of the preparation has been characterized according to the pharmacopoeieal requirements. With respect to the acute toxicity upon intragastric administration, cumazid belongs to class IV (weakly dangerous) drugs. In 3-month chronic tests, cumazid administered in doses within 1 - 100 microg/kg did not produce any significant toxic action on laboratory animals.

Pathomorphological investigation of influence on rat’s internal conditions, that were subjected to injection of medication called kumasid for prophylactic immunization extracted from Cucumaria japonica was made. It was determined that the kumasid medication dosed 1—100 mkg/kg administrated to rats intragastically during three monthes doesn’t influence on internal conditions of rats.

Fractions of the extract from meadowsweet aerial parts in 70% ethanol exhibited hepatoprotective properties during CCl 4 -induced toxic hepatitis. This extract produced a normalizing effect on activity of enzymes, markers of cytolysis, lipid peroxidation, and antioxidant defense system in liver cells. Fractionation of the extract was accompanied by dissociation of the effect. These changes reflect specific action of a complex of bioactive substances. The ethyl acetate and chloroform fractions from this extract were most potent. The effectiveness of these fractions by several parameters surpassed that of Carsil.

The extract of meadowsweet aerial parts exhibits hepatoprotective and antioxidant activity during experimental toxic CCl(4) hepatitis. This extract improved liver function. Meadowsweet extract in 70% ethanol (100 mg/kg) was most potent and exhibited low toxicity. By several parameters the effectiveness of this extract surpassed that of Carsil.

The preclinical toxicological evaluation of eplir, a new hepatoprotective drug, has been performed. It was established that, in terms of the acute toxicity parameters, eplir is a low-toxicity substance. A six-month peroral administration of the drug in a dose of 30, 150, and 300 mg/kg in rats and 100 mg/kg in dogs did not cause any substantial functional and structural disorders in the organs and systems of the experimental animals. Eplir does not possess mutagenic, carcinogenic, allergenic, and immunotoxic properties and does not affect the reproductive function.

Design of Cytochrome P-450 CYPIIB1 ligands was performed on the base of QSAR models derived by the Frontal Polygon (FP) method. The following compounds were designed: 2-phenyl-6-benzyl-2,4,6,8-tetraazabicyclo[3.3.0]octane-3,7-dione, N-acetyl-N'-(1-phenylethyl)urea, (1-phenyl-3-methylbutyl)urea. Their interaction with phenobarbital-induced hepatic microsomes from rat was investigated spectrophotometrically. The dissociation constants Ks of enzyme-substrate complexes measured are in good agreement with the values predicted by QSAR models. The results demonstrate usefulness of FP method for design of biologically active compounds.