RU2773018C2 - Cardiotropic agent with anxiolytic and analgesic properties - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to the use of trihydrochloride of N¹-(2,3,4-trimethoxybenzyl)-N²-{2-[(2,3,4-trimethoxybenzyl)amino]ethyl}-1,2-ethanediamine, which has a cardiotropic effect, as an analgesic and anxiolytic agent.

EFFECT: obtaining a cardiotropic agent with anxiolytic and analgesic properties.

1 cl, 4 ex, 3 tbl, 3 dwg

Description

Field of invention

The invention relates to medicine, in particular to pharmacology, and concerns a new application of the known substance trihydrochloride N ¹ -(2,3,4-trimethoxybenzyl)-N ² -{2-[(2,3,4-trimethoxybenzyl)amino]ethyl}- 1,2-ethanediamine (1) with cardiotropic action as an analgesic and anxiolytic agent.

State of the art

According to epidemiological data, the prevalence of anxiety and depressive disorders among patients with heart failure is significantly higher than in the general population. Approximately 13% of patients with heart failure fulfill the diagnostic criteria for an anxiety disorder, including generalized anxiety disorders and panic attacks, and almost 30% of patients have a confirmed clinically significant level of anxiety (Easton K, et al., J Cardiovasc Nurs. 2016; 31:367- 79), which is consistent with studies showing an increased risk of adverse cardiovascular outcomes in coronary artery disease comorbid with anxiety disorders. In addition, in patients with heart failure and persistent depression, a history of increased anxiety increases the risk of adverse cardiac outcomes, namely readmission and mortality (Celano CM. et al., Am Heart J. 2015; 170:1105-15).

Pain and anxiety associated with CVD are associated with long-term health risks. Preoperative anxiety and acute postoperative non-anginal pain in cardiac patients is a predictor of persistent postoperative pain up to 24 months (Choiniere M. et al., CMAJ. 2014; 186(7): Ε213-Ε223). Considering that pain and anxiety symptoms aggravate the physical and psychological state of patients with cardiovascular diseases in the postoperative period, it becomes extremely important to search for effective means and methods for relieving these symptoms.

Most psychotropic drugs used to treat mental disorders can cause side effects, the development of which is associated with their action on the somatic sphere, the effect on neurotransmitter systems and the transmission of nerve impulses, therefore, one of the contraindications for most neuropsychotropic drugs are cardiovascular diseases in the stage of decompensation .

Today, antidepressants from the group of selective serotonin reuptake inhibitors (SSRIs) remain the first-line drugs in the pharmacotherapy of depressive-anxiety disorders. The lack of proven efficacy of these antidepressants in patients with heart failure is compounded by the negative side effects of SSRIs. By inhibiting platelet aggregation/activation and increasing gastric secretion of hydrochloric acid, SSRIs may increase the risk of bleeding, especially in patients receiving antiplatelet or anticoagulant drugs (Serebruany VL. Am J Med. 2006; 119:113-6.]. SSRIs may cause lengthening (Beach SR. et al., J Clin Psychiatry. 2014; 75:e441-9) In a meta-analysis by Beach SR et al. (2014), SSRIs increased the QTc interval by an average of 6.10 milliseconds, although these changes were significantly less compared with tricyclic antidepressants (Beach SR, et al. J Clin Psychiatry. 2014; 75:e441-9) In a comparative clinical analysis of the effect of benzodiazepine and nonbenzodiazepine anxiolytics on heart rate it was shown that diazepam (7.5 mg) increased compared with placebo, and buspirone (7.5 mg), on the contrary, caused a decrease in heart rate (Unrug A. et al., Int J Psychophysio l. Feb 1997; 25(2): 177-84), which once again confirms the need for drug monitoring when prescribing anxiolytics to patients with cardiac pathology.

Oxidative damage at the cell level is currently considered as one of the mechanisms of the pathogenesis of psychological stress. According to experimental and clinical studies, there is a strong correlation between oxidative stress and the development of anxiety. For example, individuals under stress have an increase in malondialdehyde (a marker of lipid peroxidation) and abnormal levels of antioxidant enzymes in erythrocytes (Nakhaee A. et al., Physiol Behav. 2013 Jun; 118:118-21 ). Studies performed in knockout animals also demonstrate an association between increased emotional stress response and oxidative stress (Desrumaux C. et al, FASEB J. 2005 Feb; 19(2):296-7; Chen Y. et al, Neurotoxicol Teratol. 2012 Jul;34(4):450-7).

Numerous studies have noted an increase in oxidative stress in people suffering from alcoholism. So, for example, when analyzing the correlation between the severity of symptoms of acute alcohol withdrawal and markers of oxidative stress, it was found that 24 hours after the last alcohol intake in persons meeting the DSM-IV-TR criteria for the diagnosis of "alcohol dependence", there is a significant increase in the content of malonic dialdehyde and reduced serum superoxide dismutase activity compared with healthy volunteers (Huang MS et al., Prog Neuropsychopharmacol Biol Psychiatry. 2009 Feb 1; 33(1):66-9), which is consistent with previously published data by Yukel N. et al (Pharmacopsychiatry. 2005 Mar; 38(2):95-6). Moreover, the ability of the phenolic antioxidants 17β-estradiol (2), p-octyl-phenol (3), and 2,6-di-tert-butyl-4-methylphenol (4) to resist the cascade of oxidative processes induced by the abrupt withdrawal of ethanol was established. HT-22 culture of hippocampal cells exposed to ethanol, significantly reducing malondialdehyde production and protein carbonylation compared to cells without antioxidant protection (Prokai-Tatrai K et al., Int J Mol Sci. 2009; 10(4) : 1773-87)

With the introduction of ethanol (2 g/kg, orally) for 6 days, its withdrawal promoted anxiogenesis in mice. However, prior daily administration of the antioxidant flavonol quercetin (5) for 6 days prevented the development of an increased anxiety response in alcohol-deprived mice (Joshi D. et al., J Med Food. 2005 Fall; 8(3):392- 6).

Despite the lack of complete understanding of the neurobiological mechanisms underlying the effect of oxidative stress on the symptoms of anxiety disorders, it is assumed that pharmacotherapy focused on reducing the production of free radicals may have a beneficial effect on the suppression of increased anxiety of various origins.

Trimetazidine (1-[(2,3,4-Trimethoxyphenyl)methyl]piperazine dihydrochloride) was developed and marketed by the pharmaceutical company Les Laboratories SERVIE (France) as the first cytoprotective antianginal drug (6) that improves the metabolism of the myocardium and neurosensory organs under ischemia (Table 1).

Currently, trimetazidine, which has the properties of inhibitors of free fatty acid oxidation (p-FOX inhibitors), is successfully used in the treatment of coronary heart disease, chronic heart failure, metabolic syndrome and

cardiomyopathies of various origins (Heggermont WA. et al., Eur J Heart Fail. 2016; 18(12): 1420-1429), which served as an idea for the design of a new original cardiotropic compound ALM-802 (trihydrochloride N ¹ - (2,3, 4-trimethoxybenzyl)-N ² -{2-[(2,3,4-trimethoxybenzyl)amino]ethyl}-1,2-ethanediamine (1), which combines the pharmacophore elements of trimetazidine.

According to experimental studies, the compound ALM-802 has a pronounced cardiotropic effect, demonstrating antiarrhythmic activity in models of aconitine arrhythmia, calcium chloride arrhythmia and electrical fibrillation, anti-ischemic activity in a model of subendocardial ischemia (Mokroe G.V. et al., Pharmacokinetics and pharmacodynamics. 2019 No. 1. S. 6-11). Recent experimental data indicating anxiolytic (Kolik L.G. et al. Bulletin of Experimental Biology and Medicine, Vol. 162, No. 11, March, 2017 P. 642-646) and analgesic (Abdel-Salam OM, El- Batran S. Pharmacology 2005; 75:122-132) activities of trimetazidine suggest the presence of anxiolytic and analgesic properties in the spectrum of pharmacological activity of the compound ALM-802. Currently, there are no data on the analgesic and anti-anxiety effects of ALM-802 in the available literature.

The essence of the invention consists in the use of trihydrochloride N ¹ -(2,3,4-trimethoxybenzyl)-N ² -{2-[(2,3,4-trimethoxybenzyl)amino]ethyl}-1,2-ethanediamine (1) as anxiolytic and analgesic.

The purpose of the invention is the development of new agents with anxiolytic and analgesic properties. This goal was achieved by studying the pharmacological effects of ALM-802 in in vivo experiments in the "elevated plus maze" tests using genetic and pharmacological models of anxiety in different modes of administration of ALM-802, "hot plate" and "vinegar cramps".

The technical result of the invention was the expansion of the arsenal of drugs with multipotent pharmacological action, including anxiolytic and analgesic effects.

The present study was performed on inbred male mice of the BALB/c line weighing 18-20 g (n=24), male mice of the C57B 1/6 line weighing 20-24 g (n=100) and outbred white male rats (n=30) at the age of 8 months with an average weight of 450-480 g in the active phase of the experiment, obtained from the Federal State Budgetary Scientific Institution "Scientific Center for Biomedical Technologies of the Federal Medical and Biological Agency", Stolbovaya branch. The animals were kept in groups of 10 (mice) and individually (rats) in cages (370 × 200 × 150 mm) under standard vivarium conditions of the V.V. Zakusov" with adjustable light mode 12 h / 12 h (light / dark) and constant temperature (21-230C) with free access to water and granulated feed (GOST Ρ 50258-92) within 10 days before testing in accordance with the order Ministry of Health of the Russian Federation No. 199n dated August 23, 2010 "On the establishment of the Rules for Laboratory Practice." All experiments with animals were carried out in accordance with international rules (European Communities Council Directive of November 24,1986 (86/609/EEC)), as well as in accordance with the "Rules for working with animals" approved by the Bioethical Commission of the V.V. .AT. Zakusov.

ALM-802 (N1-(2,3,4-trimethoxybenzyl)-N2-{2-[(2,3,4-tri-methoxybenzyl)amino]ethyl}-1,2-ethanediamine trihydrochloride) as a substance (FGBNU Institute of Pharmacology named after V.V. Zakusov) and trimetazidine in the form of a substance (NIZHFARM JSC) as a reference drug were dissolved in water for injection and administered intraperitoneally (ip) at a rate of 0.1 ml/10 g (mice) or 0.1 ml/100 g (rat) animal weight 30 min before testing. Animals from the control group received water for injection.

To model alcohol dependence, we used the method of forced alcoholization of animals with the provision of a 10% ethanol solution as the only source of liquid for 20 weeks. The animals were kept in individual cages under standard vivarium conditions at the V.V. Zakusova" with the provision of ad libitum briquetted feed with an adjustable 12/12 light regime. Once a week, the change in body weight of the animals and the amount of ethanol consumed (in g/kg) were recorded. During forced alcoholization, rats consumed physiologically significant amounts of ethanol (4.5±0.2 g/kg in terms of ethanol). The experiments were carried out on rats with a developed stable alcohol motivation, which was assessed in terms of the average daily consumption of ethanol (per 1 kg of body weight per day) under conditions of free choice between 10% ethanol and water.

Emotional stress was modeled in the elevated plus maze test (EPM). PKL devices for mice and rats (NPK Open Science, Moscow, Russia) are made of gray plastic and have opaque side walls painted in the color of the floor. Labyrinths are raised to a height of 55 cm (for rats) and 38 cm (for mice). Each animal was placed in the center of the maze for 300 sec. Registered the following indicators: the time spent in the open sleeves; number of entries into open arms; number of entries into closed arms; time spent in the center; number of exits to the central platform; number of vertical racks; number of nods in open sleeves. The total motor activity was calculated as the sum of visits to the open and closed arms of the maze. The anxiolytic effect was assessed by the increase in the duration of stay in the open arms and the number of animal entries into the open arms of the labyrinth. The indicators “time in open arms” and “number of entries into open arms” were compared in addition to absolute values in relative units (in %), which were calculated by the formula:

Relative time in open arms, %=(time in open arms in sec / 300 sec) × 100%; relative number of entries into open arms, %=(number of entries into open arms/total number of entries into open and closed arms) x100%.

The hot plate test was used to assess the nociceptive response. Using an analgesimeter "Ugo Basile" (Italy), the latent reaction time (licking the hind paw or jumping) was recorded in C57B 1/6 mice. C57B 1/6 mice were selected 1-2 hours before the start of the experiment on the basis of basic reactivity under the conditions of this experimental model, excluding mice that remained on the plate heated to 55±0.5°C for more than 10 s. A latent period of 20 s (maximum exposure time) was regarded as 100% analgesia. The response time in mice was recorded 30, 90, and 120 min after a single injection of AL-801 at doses of 2, 4, and 8 mg/kg, ip, and trimetazidine at a dose of 8 mg/kg, ip. The choice of dose of trimetazidine is based on literature data (Abdel-Salam Μ.Ε., El-Batran S. Pharmacology 2005; 75:122-132). The results obtained were expressed as the maximum possible effect (MPE), %. MRE=(reaction latency after drug administration minus background reaction latency)/(maximum exposure time minus background reaction latency) × 100%.

The "vinegar writhing" test was performed on C57B 1/6 mice. Animals were injected with AL-802 at doses of 2, 4 and 8 mg/kg, i.p., trimetazidine at a dose of 8 mg/kg, i.p., and water for injection as control 30 min before i.p. injection 0.75 % acetic acid solution. The number of lordosis (“writhing”) in each animal was counted within 15 min after the administration of acetic acid.

The reliability of the obtained results was assessed using one-way analysis of variance (ANOVA), Duncan and Dunnett's post-hoc test, and T-test for independent groups. Data in tables and graphs are presented as mean values and standard error of means Mean ± SEM. Critical significance level α=0.05.

The invention is illustrated by the following examples

Example 1. The effect of ALM-802 after a single injection on the behavior of "highly anxious" BALB/c mice

Experiments were performed on inbred male mice of the BALB/c line with a genetically determined passive response to emotional stress (Seredenin SB, Vedernikov AA. Bull Eksp Biol Med. 1979 Jul; 88(7):38-40; Seredenin SB et al. Ann Ist Super Sanita 1990; 26(1):81-7). In the elevated plus maze (EPM) test, ALM-802, when administered acutely, prevented the formation of an anxiety reaction due to the novelty of the situation in BALB/c mice in a dose-dependent manner, statistically significantly increasing the residence time and the number of exits into the open arms (OR) labyrinth, reducing the number of entries into closed arms (CL) with no effect on general motor activity (GMA) compared with the control group (table 2). The most pronounced effect of ALM-802 was demonstrated at a dose of 2.0 mg/kg.

Example 2. The effect of ALM-802 upon chronic administration on the behavior of outbred rats under conditions of prolonged alcohol withdrawal

The experiment was carried out on male rats with alcohol dependence formed within 20 weeks and confirmed in the experiment in terms of alcohol-deprivation effect and average daily ethanol consumption (in g/kg), as well as on intact male rats who did not have access to the solution ethanol, the same age as the alcoholized animals. Alcohol deprivation for 4 weeks contributed to the formation of a pronounced anxiety reaction, expressed in a reduction in the time spent in the open arms (p<0.05) and the number of exits into the open arms (p<0.01) of the maze (table 3). ALM-802, when administered chronically systemically for 28 days at an effective cardiotropic dose of 2.0 mg/kg, prevented the decrease in the number of exits into the open arms of the LCL, induced by ethanol withdrawal, both in absolute terms (p<0.05) and in % (p<0.01), restoring these behavioral parameters to the values of intact animals (table 3).

Example 3. Influence of compound ALM-802 after a single injection on the latent period of the reaction during thermal stimulation of nociceptors

In the "hot plate" test, it was found that ALM-802, 30 minutes after administration, statistically significantly increases the reaction threshold at doses of 2.0 and 8.0 mg/kg (p<0.05, p<0.05, respectively), after 90 minutes - at doses of 4.0 and 8.0 mg/kg (p<0.01, p<0.05, respectively) and after 120 minutes at doses of 4.0 and 8.0 mg/kg (p<0.01, p<0.05, respectively), the effect persists. The dependences "dose-effect" and "time-effect" are shown, according to which the highest activity of ALM-802 after a single systemic administration is at a dose of 4.0 mg/kg 90 minutes after administration (Fig. 1). When comparing the action of ALM-802 with the reference drug trimetazidine, it was found that trimetazidine, when administered once at an effective dose of 8.0 mg/kg, exhibits moderate short-term analgesic activity (p<0.05) only 30 minutes after drug administration (Fig. 2)

Example 4. Antinociceptive effect of the compound ALM-802 in modeling tonic visceral pain in the "vinegar writhing" test

Analysis of the antinociceptive action of ALM-802 and trimetazidine in modeling tonic visceral pain in the "acetic writhing" test showed that ALM-802 at doses of 2.0 and 8.0 mg/kg with a single dose-dependently reduces the number of abdominal contractions induced by 0.75% acetic acid. acid. ALM-802 shows the highest activity at a dose of 8.0 mg/kg, surpassing the reference drug trimetazidine in terms of the severity of the effect (p<0.05) (Fig. 3)

Evidence from the scientific literature suggests many common pathophysiological mechanisms between comorbid psychiatric disorders and cardiovascular disease (eg, systemic inflammation and autonomic dysfunction). Bilateral interaction of the central nervous and cardiovascular systems can explain the growing prevalence of comorbid anxiety-depressive disorders with increasing severity of cardiovascular diseases and support the concept of alternative pathophysiological mechanisms in the presence of severe physical illness, making symptoms less sensitive or resistant to pharmacotherapy with psychotropic drugs.

For the first time, the central (anxiolytic and analgesic) effects of the original cardiotropic agent from a number of derivatives of linear methoxyphenyltriazaalkanes were revealed in the range of doses that have a positive effect on the functions of the cardiovascular system. Based on the data obtained, it can be concluded that ALM-802 can be used independently and/or as an additional remedy for the relief of anxiety caused by increased innate emotionality (an anxiety genetic model) and psycho-emotional disorders that occur during chronic alcohol dependence in conditions of alcohol abuse. deprivation (pharmacological model of anxiety), since when modeling anxiety states of various etiologies, ALM-802, with a single and chronic administration, exhibits anxiolytic activity in a standard test for assessing anxious behavior. Taking into account the fact that persons prone to alcohol abuse often develop alcoholic cardiomyopathy, which is one of the main causes of death in this category of patients, the use of ALM-802 to stop ethanol withdrawal-induced anxiogenesis in conditions of alcohol withdrawal, taking into account its main cause. cardioprotective action may be useful and effective.

Description of drawings

Fig. Fig. 1. Effect of compound ALM-802 at doses of 2.0, 4.0 and 8.0 mg/kg, i.p., on the latent period of the reaction of C57B 1/6 mice in the “hot plate” test. M±SEM

Note

On the abscissa axis - the time of development of the effect, minutes.

On the y-axis - the maximum possible effect (MPE),%.

Experimental groups:

"Control" - white rectangle;

"ALM-802 2.0 mg/kg" - light gray rectangle

"ALM-802 4.0 mg/kg" - dark gray rectangle

"ALM-802 8.0 mg/kg" - black rectangle

One-way ANOVA values (p<0.05);

*-p<0.05, **-p<0.01 compared with the corresponding control group according to Dunnett's test.

Figure 2. The effect of trimetazidine at a dose of 8.0 mg/kg, i.p., on the latent period of the reaction of C57B 1/6 mice in the "hot plate" test. M±SEM

Note:

On the abscissa axis - the time of development of the effect, minutes.

On the y-axis - the maximum possible effect (MPE),%.

Experimental groups:

"Control" - white rectangle;

"Trimetazidine 8.0 mg/kg" - gray rectangle

*-p<0.05 compared with the corresponding control group according to Student's T-test.

Fig. Fig. 3. Effect of ALM-802 (2.0 and 8.0 mg/kg, ip) and trimetazidine at a dose of 8.0 mg/kg on the number of lordosis in C57B 1/6 mice in the "acetic writhing" test. M±SEM

Notes.

On the abscissa - experimental groups:

1 - Control,

2 - ALM-802 2.0 mg/kg,

3 - ALM-802 8.0 mg/kg,

4 - Trimetazidine 8.0 mg/kg;

On the y-axis - the number of lordosis.

F(3, 46)=3.8639, p=0.01512 one-way analysis of variance (ANOVA);

*-p,0.05, **-p<0.01 compared with the control group according to Duncan's test.

Claims (1)

The use of trihydrochloride N ¹ - (2,3,4-trimethoxybenzyl) -N ² - {2- [(2,3,4-trimethoxybenzyl) amino] ethyl} -1,2-ethanediamine, which has a cardiotropic effect, as an analgesic and anxiolytic agent.

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