RU2642298C1 - Method for treatment of alcoholic cardiomyopathy - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method includes introduction of metabolitotropic cardioprotector represented by (S)-2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate in a dose of 100 mg/kg a day for 30 days. Invention application allows to increase therapeutic efficiency due to cardioprotective activity and low toxicity of (S)-2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate.

EFFECT: increased efficiency of treatment.

2 tbl,1 ex

Description

The invention relates to medicine and can be used in cardiological, therapeutic and narcological practice in the complex treatment of alcoholic cardiomyopathy (AKMP), accompanied by oxidative stress, discoordination of the thiol disulfide system, impaired myocardial histostructure, death of cardiomyocytes and the formation of heart failure.

The relevance of developing new, more advanced methods of treating alcoholic cardiomyopathy is determined by the fact that the circle of alcohol abusers remains quite wide and does not tend to decrease. So, in the countries of the former Soviet Union, including Ukraine, ethanol consumption exceeds the similar indicators of the EU countries by 50%. Chronic alcohol intoxication causes damage to almost all organs and functional systems of the body, which has led to the recent appearance of the term “alcoholic disease”. It is shown that the mortality rate of male alcoholics is 4.1, and that of women is 7.7 times higher than mortality among the rest of the population. The average age of deceased alcohol abusers is 46 years. Among the visceral manifestations of alcoholism, lesions of the cardiovascular system occupy one of the leading places, and in 30% of cases they are the direct cause of death. Currently, the works of numerous authors have established a clear relationship between alcohol abuse and pathological changes in the heart muscle, specific morphological changes in the myocardium that characterize alcoholic cardiomyopathy as an independent nosological unit are described in detail. Now a set of measures aimed at treating patients with alcoholic cardiomyopathy boils down to stopping alcohol consumption, conducting symptomatic and restorative therapy. However, this reveals the imperfection of the existing concepts of drug therapy of structurally functional myocardial disorders against the background of chronic alcohol intoxication, which makes it urgent to search and develop new, more effective methods of treating this pathology using modern highly effective pharmacotherapy.

A known method of treating alcoholic cardiomyopathy by administering to rats 2-phenyl-3-carbetoxy-4-dimethylaminomethyl-5-hydroxybenzofuran hydrochloride (vinborone) at a dose of 5 mg / kg of body weight intramuscularly 2 times a day for 14 days (Ukrainian patent No. 63731, IPC A61K 31/135 (2006.01), publication date 1/15/2004, bull. No. 1).

However, with this method of treatment, the choice of vinborone is unsuccessful, because it belongs to benzofurans, is a local anesthetic, antiarrhythmic and antispasmodic and cannot possess metabolitotropic properties ('https://vapteke.com.ua/drugs/vinboron_borschagovskyy-cpp.php). In addition, the developers offer a 14-day treatment course, which is not enough for such a pathology of the cardiovascular system. The intramuscular route of administration with a 14-day course of treatment is also not quite acceptable. 5-7 days at the injection site there are infiltrates, inflammation, etc.

впливу

кверцетину з

на

показники тварин за умов нейрогенного ушкодження

[Текст] / Ель Аараж Ахмад, I.А. Зупанець, О.О. Тарасенко // Укр. журн.

. та лаб. медицини:

науково-медичний журнал. - 2012. - Т. 7, N 3. - С. 210-214). Мы считаем неудачным выбор такой лекарственной комбинации для проведения метаболитотропной кардиопротекции, т.к кверцетин - это антиоксидант прямого типа действия, тормозящий конечные процессы оксидативного стресса, и воздействующий только на свободно-радикальные повреждения миокарда (Антиоксиданты: клинико-фармакологические аспекты / Чекман И.С., Беленичев И.Ф., Бухтиярова Н.В. и др. // Укр. мед. часопис. - 2014. - №3. - С. 1-7). Глюкозамин проявляет противовоспалительные свойства, что не позволяет заподозрить у него кардиопротективное действие в условиях АКМП.The use of quercetin in the treatment of AKMP is known (Ahmad, El Aarazh.

overwhelmingly

quercetin s

on

indicators of creatures for the brain drain of neurogenic ear

[Text] / Spruce Aarazh Ahmad, I.A. Zupanets, O.O. Tarasenko // Ukr. journal

. that lab. medicine:

medical science journal. - 2012.- T. 7, N 3. - S. 210-214). We consider it unsuccessful to choose such a medicinal combination for metabolitotropic cardioprotection, because quercetin is a direct type of antioxidant that inhibits the final processes of oxidative stress and affects only free radical damage to the myocardium (Antioxidants: clinical and pharmacological aspects / Chekman I.S. ., Belenichev I.F., Bukhtiyarova N.V. et al. // Ukrainian Medical Chronicle. - 2014. - No. 3. - S. 1-7). Glucosamine exhibits anti-inflammatory properties, which does not allow him to suspect a cardioprotective effect in conditions of AKMP.

Currently, the use in the complex therapy of alcoholic cardiomyopathy has been found by the metabolitotropic cardioprotector mildronate, which, being a structural analogue of the natural metabolite involved in oxidation-reduction processes, has a stimulating effect on energy and synthetic processes in the cell during hypoxia and toxic damage (Popov V.V. A new approach to modeling alcoholic cardiomyopathy and the effect of midronate on the restoration of structural damage to the heart muscle: Abstract of thesis. Candidate of Medical Science - Rostov-on-Don, 1990 .-- 17 p.). According to this study, mildronate was used at 50 mg / kg for 30 days after 90 days of rat alcoholization, which helped to reduce the degree of dystrophic processes in the myocardium.

A study is known “The study of the effect of alcohol and mildronate on cardiovascular activity in high altitude conditions” (Zakharov G.A., Petrov V.M., Gorokhova G.I., Ulumbekova S.A. // Bulletin of the Russian-Kyrgyz Slavic University. - 2013. - T. 13, No. 6. - S. 135-138). These studies showed the clinical inefficiency of mildronate in AKMP.

Mildronate as a metabolitotropic cardioprotector is recommended for use in case of alcoholic cardiomyopathy 500 mg intravenously 2 times a day, then orally 250 mg 4 times a day for a month (Vertkin A. Comorbid patient. A guide for practitioners. - Litres, 2016. - 140 p. .).

We consider this method a prototype.

A common essential feature of the prototype and the proposed method is the appointment for the treatment of alcoholic cardiomyopathy metabolitotropic cardioprotector.

However, in real clinical practice, when prescribing Mildronate, it is not always possible to achieve an adequate clinical effect due to the fact that Mildronate is not a strictly targeted drug for alcoholic cardiomyopathy, it does not fully affect the intimate mechanisms of the damaging effect of ethyl alcohol on the myocardium.

Therapy according to the prototype method has a certain metabolitotropic cardioprotective effect in AKMP, however, it is weaker than in the treatment according to the claimed method.

The basis of the invention is the task of improving the method of treatment of alcoholic cardiomyopathy by introducing into the treatment regimen a medicinal product containing, as the active substance, (S) -2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate, which, influencing directly the mechanisms of the damaging effect of alcohol on the myocardium, it has a pronounced metabolitotropic cardioprotective effect and has low toxicity, which will increase the effectiveness of treatment.

The problem is solved in that in the method of treating alcoholic cardiomyopathy by prescribing a metabolitotropic cardioprotector, a new one is that as a metabolitotropic cardioprotector, a drug is prescribed containing the active substance - (S) -2,6-diaminohexanoic acid 3-methyl-1, 2,4-triazolyl-5-thioacetate in a dose of 100 mg / kg per day for 30 days.

A causal relationship between the totality of the claimed features and the technical result is as follows.

Despite the fact that alcoholic cardiomyopathy is clearly described and since 1996 has been considered by WHO as part of dilated cardiomyopathy, and according to the ICD, it has been singled out in a separate nosological form (I42.6), there are certain difficulties in the practical formulation of this diagnosis and the creation of specific treatment regimens for this pathology. Effective treatment regimens for AKMP taking into account the damaging mechanisms of the effects of alcohol on the heart have not been developed to date.

AKMP refers to secondary toxic dilated cardiomyopathy (DCMP). According to estimates of various authors, in 2-36% of cases, DCMP has an alcoholic etiology, in which case it should be considered as part of AKMP. A feature of the formation of AKMP, unlike other cardiomyopathies, is alcohol-dependent damage to the mitochondria of the myocardium, which makes mitochondria a source of reactive oxygen species and proapoptotic proteins, and against the background of worsening energy production (decreased ATP), oxidative stress and apoptosis are activated. In addition, several more links of the cardiodestructive effect of ethanol are distinguished - the direct toxic effect of acetaldehyde and ethanol on protein synthesis, the breakdown of the conjugation between excitation and contraction, lipid metabolism and the formation of fatty degeneration, an imbalance of catecholamines and other hormones, ionic imbalance, the effect on the cytoskeleton, activation proviruses, changes in the processes of excitation and conduction in the heart muscle (Aberle N. et al. Experimental Assessment of the Role of Acetaldehyde in Alcoholic Cardiomyopathy // Biol. Proced. Online. 2003; 5: 1-12). The mechanisms highlighted above, realizing their harmful effects on the heart, ultimately lead to the development of heart failure. A fundamentally important process is heart remodeling. This concept includes a violation of the structure of the contractile apparatus of cardiomyocytes, their functional asymmetry, changes in intercellular interactions, interstitial fibrosis, despiralization of the muscle bundles and changes in the shape of the heart cavities (Drapkina, O. Problem of alcoholic cardiomyopathy / O. Drapkina, Y. Ashikhmin, V. Ivashkin // Doctor. - 2005. - No. 8. - S. 48-50).

Based on this, against the background of basic therapy, which includes beta-blockers, ACE inhibitors, calcium channel blockers, antiplatelet agents, metabolitotropic therapy should deserve special attention. Metabolitotropic therapy in AKMP, in contrast to that in chronic heart failure, should be aimed at maintaining the structural and functional features of myocardial mitochondria, i.e. carry elements of mitoprotection. Comprehensive metabolitotropic cardioprotection in AKMP should restore metabolic alcohol-dependent shifts in the biochemical continuum of cardiomyocytes so that each previous drug ingredient creates substrates for the action of the subsequent medication.

In addition, it is considered promising to use in the treatment of AKMP as first-line cardioprotectors drugs that switch myocardial metabolism using fatty acids to aerobic breakdown of glucose (as a more effective way), inhibiting mitochondrial ketoacyl-CoA thiolase, as well as normalizing the Krebs cycle, regulating mitochondria -cytosolic compensatory energy shunts (in particular, malate-aspartate).

And in this regard, our interest was caused by the derivative of 1,2,4-triazolyl-5-thione - (S) -2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate (conventional name Angiolin) , which exhibits a cardioprotective effect in experimental myocardial infarction and a neuroprotective, metabolitotropic, endothelioprotective effect in cerebral ischemia and chronic alcohol intoxication (Ukrainian patent No. 106867 "Method for the correction of neurological disorders in chronic alcohol intoxication", publ. 10.10.2014, bull. No. 19 ; patent of Ukraine 86668 "LYSINE 3-methyl-1,2,4-triazolyl-5-thioacetate", publ. 12.05.2009, Bul. №9), but its action as metabolitotropnogo cardioprotector at AKMP until now it was not known and was not investigated.

In the course of experimental studies, we obtained data on the metabolitotropic cardioprotective effect of Angiolin in alcoholic cardiomyopathy, which is aimed at improving myocardial histostructure, inhibition of oxidative stress and normalization of the myocardial thiol disulfide system, preserving the structural and functional characteristics of myocardial mitochondria (mitoprotection, i.e. action directly positively affecting the pathogenetic mechanisms of the development of alcoholic cardiomyopathy.

Below we provide data that confirm this.

Example.

In these studies, the optimal dose of Angiolin for oral administration, which is 100 mg / kg, was determined experimentally. Based on this, when conducting the experiments described below, we used the indicated dose.

The experiment used 40 white mongrel white male rats weighing 170-180 g, obtained from the nursery of the Institute of Pharmacology and Toxicology of the Academy of Medical Sciences of Ukraine. Alcoholic cardiomyopathy was caused by intragastric administration of a 25% ethanol solution at a dose of 6 g / kg with a metal probe for 90 days. After cessation of animal administration of alcohol, Angiolin was administered intragastrically for 30 days at a dose of 100 mg / kg and the reference drug Mildronate at a dose of 250 mg / kg. At the end of the experimental therapy, the animals were removed from the experiment under thiopental sodium anesthesia (40 mg / kg), and their heart was removed. The apical part of the heart was placed in Carnoy's fixative for 24 hours. After the standard procedure of tissue dehydration and its impregnation with chloroform and paraffin, the myocardium was poured into a paraplast (MkCormick, USA). Using a Microm-325 rotational microscope (Microm Corp., Germany), serial histological sections 5 μm thick were prepared. To study the morphological and functional state of the myocardium and nuclei of endotheliocytes, the histological sections were dewaxed according to the standard method and stained with halocyanine-chromium alum according to Einarson for specific detection of RNA. Morphometric analysis of cardiomyocytes was carried out on an Axioskop microscope (Ziess, Germany) using a COHU-4922 video camera (USA) and introduced into the VIDAS-386 digital image analysis system (Kontron Elektronic, Germany). The following indicators were determined: the area of the nuclei of cardiomyocytes (μm ² ); the concentration of RNA in the nuclei of cardiomyocytes in units of optical density (E _OD ); the concentration of RNA in the cytoplasm of cardiomyocytes in units of optical density (E _OD ); density of cardiomyocyte nuclei as an indicator of the number of cell nuclei per 1 mm ^{2 of} myocardial tissue area; nuclear cytoplasmic coefficient as an indicator of the total area of cardiomyocyte nuclei per 1 mm ^{2 of} myocardial tissue area.

For biochemical studies, the heart was washed with a cooled solution of 0.15 M KCl (4 ° C) 1:10, and then ground in liquid nitrogen to a powder state and stirred in a 10-fold volume of medium at (2 ° C) containing (in mmoles) : sucrose 250, Tris-HCl buffer 20, EDTA 1 (pH 7.4). At a temperature of + 4 ° С, a cytosolic fraction was isolated by differential centrifugation on a Sigma 3-30k refrigerator centrifuge (Germany). In the cytosol, the oxidative stress marker nitrotyrosine was determined, as well as indicators of the glutathione unit of the thiol disulfide system of the myocardium — reduced glutathione and glutathione peroxidase activity (GSR).

As can be seen from the data presented in table 1, in rats with AKMP there was a significant decrease in the nuclear cytoplasmic coefficient by 26.2% and nuclear density by 19.1% compared with intact animals, which reflects the presence of pathological myocardial hypertrophy, apoptosis and death of cardiomyocytes. An increase in the area of the nuclei of cardiomyocytes by 10.5% was also observed, with a parallel decrease in the concentration of RNA in the nuclei by 24% and in the cytoplasm of cardiomyocytes by 26.6%. Taking into account the important role of RNA in the synthesis of structural and enzymatic proteins of cells, the possibility of suppressing the protein-synthetic function of cardiomyocytes in conditions of alcoholic cardiomyopathy becomes apparent. Thus, the morphological changes in cardiomyocytes in rats with AKMP are characterized by a decrease in the density of cardiomyocyte nuclei, an increase in their area, a decrease in the concentration of nuclear and cytoplasmic RNA, and a low nuclear cytoplasmic coefficient compared to intact animals, i.e. characterized by signs of pathological myocardial hypertrophy. The administration of Angiolin in a dose of 100 mg / kg for 30 days after a 3-month administration of ethanol also had a cardioprotective effect - it led to a reliable, compared with the control group, increase in the density of cardiomyocyte nuclei by 18.1% (this indicates a cardioprotective effect of the drug) , an increase of 26.3% in the concentration of RNA in the nuclei of cardiomyocytes and an increase of 45.4% in the concentration of RNA in the cytoplasm of cardiomyocytes (indicates an increase in translational processes in cardiomyocytes and stimulation of protein synthesis). The introduction of Angiolin led to a normalization of the area of cardiomyocytes (a decrease of this indicator compared to the control by 12.2%), indicating a decrease in myocardial hypertrophy. Angiolin in a dose of 100 mg / kg significantly increased by 20% the nuclear cytoplasmic index of rat myocardium with AKMP, which indicates the presence of antihypertrophic effect of this drug. The course administration of Mildronate at a dose of 250 mg / kg led to a greater, compared with the control group, decrease in the density of cardiomyocyte nuclei (by 6.9%) and a decrease in the nuclear cytoplasmic coefficient by 6.6% against the background of an increase in the area of the nuclei of cardiomyocytes by 9, 5%. The revealed data indicate that Mildronate not only did not affect the signs of pathological myocardial hypertrophy with alcoholic cardiomyopathy, but also exacerbated alcoholic heart damage. The introduction of Mildronate led to a significant increase in RNA in the cytoplasm of cardiomyocytes by 18%, without affecting this indicator in the nuclei. By the strength of the cardioprotective effect, Angiolin significantly exceeds the reference drug Mildronate (250 mg / kg) in all indicators of the morphological and functional characteristics of the myocardium under conditions of AKMP.

As can be seen from the data presented in table 2, the simulation of alcoholic cardiomyopathy (AKMP) leads to a sharp surge in free-radical reactions of oxidative stress against the background of inhibition of the antioxidant (thiol-disulfide) system. Thus, in the myocardium of animals with AKMP, a 2-fold decrease in the level of reduced glutathione was observed against a background of a 3-fold decrease in the activity of the glutathione-dependent enzyme, HSP, and a significant increase in the myocardium of the oxidative stress marker nitrotyrosine was recorded by a factor of 12. The course administration of Angiolin in animals with AKMP led to inhibition of oxidative stress in the myocardium, as evidenced by a decrease in nitrotyrosine by 78.1%. In parallel, in the myocardium of animals with AKMP treated with Angiolin, an increase in the intermediates of the glutathione link of the thiol-disulfide system was recorded - an increase in the concentration of reduced glutathione by 76% and an increase in the activity of HPR by 143.2%. The course administration to animals with AKMP Mildronate did not have a significant effect on the level of reduced glutathione, did not have a pronounced effect on the increase in the activity of HGP (+ 28.3%) and on the decrease in the content of nitrotyrosine (-14.3%) in the myocardium. Angiolin significantly exceeds Mildronate in terms of its effect on indicators such as an increase in the content of reduced glutathione, the activity of HGP, and a decrease in the level of nitrotyrosine.

The molecular biochemical mechanisms of the development of alcoholic cardiomyopathy have their own peculiarities, therefore, drugs that are quite effective in various other types of cardiomyopathies can be ineffective in AKMP. The metabolitotropic cardioprotective activity of Angiolin is aimed at eliminating structural and molecular biochemical disturbances in alcoholic myocardial damage, this was not obvious to a specialist, and was revealed as a result of our experimental studies. This effect of Angiolin was higher in comparison with known drugs, in particular, in comparison with the reference drug Mildronate. Based on the foregoing, we can assume that our method is new, has an inventive step and can be used with high efficiency in the treatment of alcoholic cardiomyopathy.

Claims (1)

A method for the treatment of alcoholic cardiomyopathy by administering a metabolitotropic cardioprotector, characterized in that a drug containing 3-methyl-1,2,4-triazolyl-5-thioacetate as an active substance (S) -2,6-diaminohexanoic acid is prescribed as a metabolitotropic cardioprotector at a dose of 100 mg / kg per day for 30 days.