RU2526902C1 - Agent having cardioprotective action and 1,3-disubstituted 2-aminobenzimidazolium halides - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to pharmaceutics and medicine and use of tricyclic imidazo[1,2-a]benzimidazole derivatives of general formula I

or 1,3-disubstituted 2-aminobenzimidazolium halides of general formula II

as an agent having cardioprotective action with high efficiency.

EFFECT: high efficiency.

3 cl, 1 tbl, 2 ex

Description

The invention relates to agents having a cardioprotective effect.

The genesis of cardiomyocyte damage in a number of diseases of the cardiovascular system is based on the acidification of the intracellular environment and the development of the “calcium paradox”. These changes, due to various etiological factors, acquire important pathogenetic significance and are often crucial for the prognosis of the disease. In this regard, the desire to exert a corrective effect on the disturbed metabolism of cardiomyocytes [Mikhin V.P., Savelyeva V.V. is natural The role of cardiocytoprotectors in the treatment of chronic heart failure of ischemic origin. / Russian Journal of Cardiology. - 2009. - No. 1 (75). P.49-56].

In recent years, as a possible physiological and pathophysiological factor in the development mechanisms of such consequences of ischemic and reperfusion stress, such as arrhythmias, impaired contractile function and cell death, sarcolem Na ⁺ / H ⁺ exchanger 1 isoforms (NHE-1) is considered [O. Pisarenko O. AND. Na ⁺ / H ⁺ exchange inhibitors are a new class of cardioprotectors. / Russian physiological journal. THEM. Sechenov. - 2004. - No. 9 (90). - S.1103-1110].

Its biological role is associated with processes such as the exchange of Na ⁺ and H ⁺ ions; regulation of pH and cell volume; growth, hypertrophy, differentiation; cell damage with ischemia; in the organization of the cytoskeleton and cell migration [Slepkov ER, Rainey JK, Sykes BD, Fliegel L. Structural and functional analysis of the Na ⁺ / H ⁺ exchanger. / Biochem. J. - 2007. - No. 401 (3). - R.623-633].

NHE1 activation occurs during ischemia and reperfusion. When ischemia Na ⁺ / H ⁺ exchanger starts to output cells of sarcoplasmic H ⁺ ions accumulated there due to the activation of glycolysis process in exchange for the input of ions Na ^+. An increase in the intracellular content of Na ⁺ ions in turn activates the 3Na ⁺ / Ca ²⁺ -version exchange (NCE exchanger) through the sarcolemma, initiating the entry of Ca ^{2+ ions} into cardiomyocytes. There is a "Ca ²⁺ overload." Reperfusion exacerbates the damage that occurred during ischemia. During reperfusion, the concentration of extracellular protons decreases rapidly, a large gradient of extra / intracellular protons is formed, which leads to an increase in calcium levels. As a result, contractile function deteriorates, severe rhythm disturbances develop, which ultimately leads to the death of cardiomyocytes. The mechanism of action of NHE inhibitors is ultimately associated with the prevention of this calcium overload [Fliegel L. Regulation of the Na (+) / H (+) exchanger in the healthy and diseased myocardium. Expert. Opin. Ther. Targets. - 2009. - No. 13 (1). - R. 55-68].

The main positive effects of NHE inhibitors on ischemic myocardium in experimental and clinical studies are associated with improved recovery of contractile function of the left ventricle after reperfusion; reduction of the area of ischemia and necrosis; a decrease in the severity of the violation of ionic homeostasis accompanying ischemia / reperfusion; reduction of apoptosis, a decrease in the incidence and severity of arrhythmias caused by ischemia / reperfusion.

A cardioprotective agent known among NHE inhibitors is Zoniporide (CP-597396) - [1- (quinolin-5-yl) -5-cyclopropyl-1H-pyrazole-4-carbonyl] guanidine hydrochloride monohydrate [A Guzman-Perez et al . Discovery of zoniporide: A potent and selective sodium-hydrogen exchanger type 1 (NHE-1) inhibitor with high aqueous solubility ./ Bioorganic & Medicinal Chemistry Letters. - 2001. - No. 11 (6). - R.803-807]. In vitro studies on rabbit platelets have shown that this drug inhibits NHE-1-dependent platelet swelling (maximum inhibition of 93%) [Tracey WR et al. Zoniporide: a potent and selective inhibitor of the human sodium-hydrogen exchanger isoform 1 (NHE-1) // Cardiovasc. Drug Rev. - 2003. - No. 21. - R.17-32]. In studies on an isolated heart according to Langendorff, zoniporide causes a dose-dependent decrease in infarct size (EC ₅₀ = 0.25 nM). At a dose of 50 nM, the drug reduces the size of myocardial infarction by 83%, which is 2.5 times higher than eniporide or cariporide (EC ₅₀ 0.69 and 5.11 nM, respectively). An in vivo model of ischemic myocardial damage showed that zoniporide has a cardioprotective effect [Knight DR et al. A novel sodium-hydrogen exchanger isoform-1 inhibitor, zoniporide, reduces ischemic myocardial injury in vitro and in vivo // J. PharmacoI.Exp.Ther. - 2001. - No. 297 (1). - P.254-259]. Zoniporide markedly reduced the size of the heart attack without impaired hemodynamics and contractile function of the heart, weakened post-reperfusion contractile dysfunction, and also reduced the frequency and duration of ventricular fibrillation [Tracey WR et al. Zoniporide: a potent and selective inhibitor of the human sodium-hydrogen exchanger isoform 1 (NHE-1) // Cardiovasc Drug Rev. - No. 21. - 2003. - P.17-32]. However, it did not significantly affect coronary blood flow, and its main effect was associated with metabolic changes [Ayoub IM et al. Zoniporide preserves left ventricular compliance during ventricular fibrillation and minimizes postresuscitation myocardial dysfunction through benefits on energy metabolism // Crit Care Med. - 2007. - No. 35 (10). - P. 2329-2336].

Clinical trials of the drug were conducted by Pfazer in patients with myocardial infarction, coronary artery disease, atherosclerosis. The most common side effects in patients receiving zoniporide were drowsiness and orthostatic hypotension, and nausea, vomiting, and headache were also observed [Dalvie D. et al. Cross-species comparison of the metabolism and excretion of zoniporide: contribution of aldehyde oxidase to interspecies differences / Drug Metab. Dispos. 2010 - No. 38 (4). - P.641-654]. Therefore, the neurotoxic effect of zoniporide was studied in rats, both clinical and electrophysiological microscopic and microscopic properties of peripheral sensory axonopathies [Pettersen JC et al. Neurotoxic effects of zoniporide: a selective inhibitor of the Na ⁺ / H ⁺ exchanger isoform 1 // Toxicol Pathol. - 2008. - No. 36 (4). - P.608 -619]. The risk of damage to the peripheral nervous system may limit the dose and duration of therapy for cardiovascular disease [Liu CN, Somps CJ 1Na ⁺ / H ⁺ Exchanger-1 Inhibitors Reduce Neuronal Excitability and Alter Na ⁺ Channel Inactivation Properties in Rat Primary Sensory Neurons. / TOXICOLOGICAL SCIENCES. - 2008. - No. 103 (2). - P.346-353].

The technical result is to increase the effectiveness of the cardioprotective action.

The technical result is achieved by using, as a cardioprotective agent, tricyclic derivatives of imidazo [1,2-a] benzimidazole of the general formula I:

where Ar = C ₆ H ₄ OCH ₃ -p, C ₆ H ₄ OH-p

NR ₂ = N (CH ₂ CH ₂ ) ₂ O,

including

Ia Ar = C ₆ H ₄ OCH ₃ -p; NR ₂ = N (CH ₂ CH ₂ ) ₂ O

Ic Ar = C ₆ H ₄ OH-p; NR ₂ = N (CH ₂ CH ₂ ) ₂ O

or halides of 1,3-disubstituted 2-iminobenzimidazolium of the general formula I:

where Ar = C ₆ H ₄ F-4, C ₆ H ₃ (O ₂ CH ₂ ) -3.4

NR ₂ = N (C ₂ H ₅ ) ₂ , N (CH ₂ CH ₂ ) O; X = Cl, Br,

including

IIa Ar = C ₆ H ₄ F-4; NR ₂ = N (CH ₂ CH ₂ ) ₂ O, X = Cl

IIc Ar = C ₆ H ₃ (O ₂ CH ₂ ) -3.4; NR ₂ = N (C ₂ H ₅ ) ₂ , X = Br.

The compounds can be used, inter alia, for the manufacture of agents having a cardioprotective effect, as well as for the restoration and improvement of cardioprotective functions in living organisms.

Compound Ia is known to be able to inhibit cAMP PDE and reduce atrial excitability (V.A. Anisimova, A.A. Spasov, V.A. Kosolapov, A.F. Kucheryavenko, O.V. Ostrovsky, N.P. Larionov, RE Libinson, Pharmacological Activity of 2-Methoxyphenyl-Substituted 9-Dialkylaminoethylimidazo [1,2-a] Benzimidazoles, Chemical Pharmaceutical Journal, 2005, vol. 39, No. 9, pp. 26-32) properties of an antagonist of purine P ₂ Y ₁ receptors and antithrombotic activity [RF patent No. 2377990, IPC A61K 31/5377, 2006).

Compound Ib exhibits antioxidant and antiplatelet activity, as well as a weak antiserotonin and antipurinergic effect (V.A. Anisimova, A.A. Spasov, V.A. Kosolapov, M.V. Chernikov, A.Yu. Stukovina, L.V. Eltsova, N.P. Larionov, R.E. Libinson, and O.E. Vatolkina, Synthesis and Biological Activity of 9-Dialkylaminoethyl-2-hydroxy (dioxi) phenylimidazo [1,2-a] benzimidazoles, Chemical Pharmaceutical Journal , 2006, vol. 40, No. 10, pp. 23-26).

Compounds IIa, IIc are not described.

The technical result is also new compounds in the series of 1,3-disubstituted 2-aminobenzimidazolium halogenides having a cardioprotective effect.

The technical result is achieved by the halides of 1-dialkylaminoethyl-3- [substituted (disubstituted) phenacyl] -2-aminobenz-imidazolium of the general formula II:

where IIa Ar = C ₆ H ₄ F-4; NR ₂ = N (CH ₂ CH ₂ ) ₂ O, X = Cl

IIc Ar = C ₆ H ₃ (O ₂ CH ₂ ) -3.4; NR ₂ = N (C ₂ H ₅ ) ₂ , X = Br.

The following are examples of the preparation of compounds IIa and IIc.

Example 1. Synthesis of chloride of 1-morpholinoethyl-3- (4-fluorophenacyl) -2-aminobenzimidazolium (IIa).

A mixture of 1.23 g (5 mmol) of 1-morpholinoethyl-2-aminobenzimidazole, 0.87 g (5 mmol) of ω-chloro-4-fluoroacetophenone and 15 ml of dry acetonitrile is boiled until the quaternization reaction is complete (~ 8-10 hours, control - TLC). Then acetonitrile is evaporated to a small volume, the precipitate, which is the desired chloride, is filtered off and washed with acetone. Yield 92.5%. Mp 198-199 ° C.

Found,%: C 60.15; H 5.68; Cl 8.54; F 4.60; N, 13.45. C ₂₁ H ₂₄ ClFN ₄ O ₂ .

Calculated,%: C 60.21; H 5.77; Cl 8.46; F 4.54; N, 13.37.

IR spectrum, v, cm ^-1 : 3206, 3168, 3022 (N ⁺ H ₂ ), 1693 (C = O), 1663 (C = N), 1599 (C = C); 1226 (CN); 1118 (CO).

¹ H NMR Spectrum (DMSO-d ₆ ), δ, ppm (J, Hz): 2.40-2.60 m [4H, N (CH ₂ ) ₂ ], 2.67 t (2H, J = 5.7 Hz, NCH ₂ ), 3.48 t [4H, J = 3.9 Hz, O (CH ₂ ) ₂ ], 4.39 t (2H, J = 5.7 Hz, NCH ₂ ), 6.05 s (2H, CH ₂ ), 7.24-7.62 m (6H, H _Ar ), 8.14-8.21 m (2H, H _Ar ), 9.22 br s (2H, N ⁺ H ₂ ).

Example 2. 1-Diethylaminoethyl-2-amino-3- (3,4-methylenedioxyphenacyl) benzimidazolium bromide (IIc).

1.22 g (5 mmol) of 3,4-methylenedioxyphenacyl bromide are added to a hot solution of 1.16 g (5 mmol) of 1-diethylaminoethyl-2-aminobenzimidazole in acetone, the mixture is stirred and left for 3-4 hours at 20-25 ° C . The precipitated bromide precipitate is separated and washed with acetone. Yield 97.3%. Mp 235-236 ° C (decomposition).

IR spectrum (solid state), v, cm ^-1 : 3310, 3236.3090, 1681, 1652, 1449, 1255, 1033.751.

¹ H NMR spectrum (DMSO-d ₆ ), δ, ppm: 0.81 t (6H, J 6.9 Hz, 2CH ₃ ); 2.73 t (2H, J 6.2 Hz, CH ₂ ); 4.29 t (2H, J 6.2 Hz, CH ₂ ); 5.88 s (2H, CH ₂ CO); 6.20 s (2H, OCH ₂ O); 7.15-7.40 m (3H, H _Ar ), 7.48-7.65 m (3H, H _Ar ), 7.75 d (1H, J 8.1 Hz, H _Ar ), N ⁺ H ₂ in exchange.

Below are the test results of the cardioprotective properties of the compounds.

1. Materials and methods.

1.1. Materials

Compounds I and II, zoniporide (SIGMA, USA).

Reagents: sodium chloride “ChCH” NaCl (ZAO NPO ECROS, Russia), a set of reagents for enzyme-linked immunosorbent assay Rat cardiac troponin I (cTn-I) (cat. No. CSB-E08420r) from CUSABIO BIOTECH CO., LTD. (China), Evans blue, triphenyltetrazolium, Evans blue (SIGMA, USA), 2,3,5-triphenyltetrazolium chloride (SIGMA, USA).

1.2. Animals.

In the experiments, 57 female rats weighing 260-300 g were used, which were kept under vivarium conditions with natural light on a standard diet of laboratory animals in accordance with GOST P 50258-92 [1993]. The studies were carried out in accordance with the applicable requirements of GOST ISO / IEC 17025-2009, GOST P ISO 5725-2002 and the "Laboratory Practice Rules", approved by order of the Ministry of Health and Social Development of the Russian Federation of August 23, 2010 No. 708n, in compliance with the "European Convention for the Protection of Vertebrate Animals Used for experiments or other scientific purposes ”[Directive 2010/63 / EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes Official Journal L 276, 10.20.2010, p. 33-79 (revising Directive 86/609 / EEC)].

Animals were divided into 11 groups:

1-3 control:

1 - “control-falsely operated” - the animals underwent a full range of operations, except ligation of the left coronary artery;

2 - “control-ischemia / reperfusion” - the animals underwent ligation of the left coronary artery with subsequent reperfusion, physiological saline was injected;

3 - "control zoniporide" - animals with ligation of the left coronary artery were introduced zoniporide 10 minutes before reperfusion.

4-7 experimental (animals with ligation of the left coronary artery were injected with the studied compounds 10 minutes before reperfusion):

4 - "IIa"; 5 - "Ia"; 6 - "IIv"; 7 - "Iv."

1.3. Methods

The experiments were performed on anesthetized female rats (chloral hydrate, 400 mg / kg) under conditions of artificial ventilation, thoracotomy, pericardotomy. Ischemia / reperfusion was modeled by 60 minutes of ligation of the left coronary artery, followed by 60 minutes of reperfusion. Saline, zoniporide, the studied compounds were administered intravenously in equivalent volumes of 1 mg / kg 10 minutes before reperfusion. The development of myocardial ischemia was monitored by an electrocardiogram in standard II lead (Poly-Spectrum-8 / V computer electrocardiograph, Russia); the size of the anatomical risk zone and myocardial necrosis, myocardial damage marker - troponin I rats.

The dimensions of the anatomical risk zone and infarction zone were estimated using the double staining technique with blue Evans and triphenyltetrazolium followed by calculation in the Image J.

The plasma plasma troponin I content as a marker of myocardial damage was determined by immunochemical reagent assay kit Rat cardiac troponin I (cTn-I) from CUSABIO BIOTECH CO., LTD. (China) on the universal microplate reader ELX 800 manufactured by Bio-Tek Instruments, Inc (USA). The troponin I content was expressed in PCG / ml plasma.

1.4. Statistical Processing Methods

All statistical calculations were performed using the Statistica for Windows 6.0 software package, StatSoft, Inc. (USA). A pairwise comparison of the samples was performed using the Mann-Whitney U test. The hypothesis of the existence of differences between the samples was accepted at a level p <0.05.

2. The results of the study

When studying the effect of compounds on the size of the risk zone and myocardial infarction under conditions of ischemia / reperfusion, the following results were shown. In animals of group 2–7, during ligation of the left coronary artery, ischemic damage to the left ventricular myocardium developed, which was confirmed by the elevation of the ST segment on the ECG in standard lead II and when stained with blue Evans. In the control and experimental sections, the risk zone in all groups averaged 42-55% (table 1). This confirms the uniformity of the surgical interventions.

Subsequent staining with triphenyltetrazolium in the control-ischemia / reperfusion group, the necrosis zone was 44.36%. With the introduction of zoniporide, IIa, IIc, Ia, Ic, there is a significant decrease in the zone of myocardial infarction compared with the control-ischemia / reperfusion group. Moreover, the studied compounds exceeded zoniporide in their activity (table 1). Compound IIa is statistically significantly 1.4 times superior to the reference drug.

When analyzing changes in the level of troponin I in the blood plasma of animals of the control-ischemia / reperfusion group, a significant increase was shown in comparison with the falsely operated ones (table 1.). In the experimental groups, a statistically significant decrease in the damage marker in relation to the control-ischemia / reperfusion group was observed with the introduction of zoniporide, compounds IIa, Ia, IIc, Ic in 2.1, 2.5; 1.7; 1.9 and 1.5 times, respectively.

Table 1 shows the main indicators of the cardioprotective effect of compounds Ia, Ib and IIa, IIc in the ischemia / reperfusion model when administered intravenously at a dose of 1 mg / kg.

Table 1 The main indicators of the cardioprotective effect on the model of ischemia / reperfusion with intravenous administration at a dose of 1 mg / kg No. Group n Dimensions of the risk zone (% in relation to the left ventricle) The size of the necrosis zone (% in relation to the risk zone) The level of troponin I (PCG / ml) one. Control-falsely operated 6 0.00 ± 0.00 0.00 ± 0.00 136.65 ± 38.03 2. Ischemia Control / Reperfusion 7 53.20 ± 4.99 44.36 ± 3.79 528.32 ± 81.32 ^# 3. Control zoneiporide 7 47.25 ± 3.82 34.87 ± 2.57 ^* 252.76 ± 72.05 ^* four. IIa 5 46.33 ± 3.2 24.10 ± 1.47 ^{* $} 211.88 ± 60.14 ^* 5. Ia 8 45.01 ± 1.77 27.78 ± 4.23 ^* 316.55 ± 62.34 ^* 6. IIc four 51.09 ± 6.08 29.92 ± 4.84 ^* 287.46 ± 60.14 ^* 7. Ic 5 46.04 ± 5.28 30.04 ± 2.58 ^* 346.78 ± 99.75 ^{* #} - the value is statistically significant in relation to the “control-falsely operated” group; * - the value is statistically significant in relation to the control-ischemia / reperfusion group; ^$ - the value is statistically significant in relation to the control-zoniporide group.

3. Conclusions

Thus, in vivo studies on the ischemia / reperfusion model showed the cardioprotective effect of the studied benzimidazole derivatives of compounds Ia, Ib and IIa, IIc.

It was shown that compounds Ia, Ic, IIc, when administered intravenously at a dose of 1 mg / kg, statistically significantly in relation to the control-ischemia / reperfusion group reduce the size of the myocardial necrosis zone and the level of troponin I in the blood in the rat ischemia / reperfusion model, having an effect similar to zoniporid.

Compound IIa, when administered at a dose of 1 mg / kg, was statistically significantly superior to zoniporide in terms of its effect on the size of the myocardial necrosis zone and reduced the level of troponin I in the blood to a greater extent on the rat ischemia / reperfusion model.

Claims (3)

1. The use as a cardioprotective agent of tricyclic derivatives of imidazo [1,2-a] benzimidazole of the general formula I:

where Ar = C ₆ H ₄ OCH ₃ -p, C ₆ H ₄ OH-p
NR ₂ = N (CH ₂ CH ₂ ) ₂ O,
including
Ia Ar = C ₆ H ₄ OCH ₃ -p; NR ₂ = N (CH ₂ CH ₂ ) ₂ O
Ic Ar = C ₆ H ₄ OH-p; NR ₂ = N (CH ₂ CH ₂ ) ₂ O
or halides of 1,3-disubstituted 2-aminobenzimidazolium of the general formula II:

where Ar = C ₆ H ₄ Fp, C ₆ H ₃ (O ₂ CH ₂ ) -3.4
NR ₂ = N (C ₂ H ₅ ) ₂ , N (CH ₂ CH ₂ ) OX = Cl, Br,
including
IIa Ar = C ₆ H ₄ Fp, NR ₂ = N (CH ₂ CH ₂ ) ₂ O, X = Cl
IIc Ar = C ₆ H ₃ (O ₂ CH ₂ ) -3.4, NR ₂ = N (C ₂ H ₅ ) ₂ , X = Br. 2. The use according to claim 1 for the manufacture of agents having a cardioprotective effect, as well as for the restoration and improvement of cardioprotective functions in living organisms. 3. Halides of 1,3-disubstituted 2-aminobenzimidazolium of the general formula II:

where Ar = C ₆ H ₄ F-p, C ₆ H ₃ (O ₂ CH ₂ ) -3.4
NR ₂ = N (C ₂ H ₅ ) ₂ , N (CH ₂ CH ₂ ) OX = Cl, Br,
including
IIa Ar = C ₆ H ₄ Fp, NR ₂ = N (CH ₂ CH ₂ ) ₂ O, X = Cl
IIc Ar = C ₆ H ₃ (O ₂ CH ₂ ) -3.4, NR ₂ = N (C ₂ H ₅ ) ₂ , X = Br.