RU2645356C1 - 3-(1h-benzimidazol-2-yl) -1,2,2-trimethyl cyclopentancarbonic acid with cardioprotective activity - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: invention relates to application of 3-(1H-benzimidazol-2-yl)-1,2,2-trimethylcyclopentanecarboxylic acid as an agent having cardioprotective activity.

EFFECT: new properties of this acid are revealed that allow to increase the effectiveness of cardiovascular diseases treatment.

1 tbl, 1 ex

Description

The invention relates to the field of pharmacology, namely to biologically active compounds, in particular 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid, C ₁₆ H ₂₀ N ₂ O ₂ exhibiting cardioprotective activity.

According to well-known literature, the cardiotoxicity of chemotherapy is one of the main causes of mortality in cancer patients, primarily for children, patients with hemablastoses, patients with breast cancer and Hodgkin’s lymphoma [Felker GM, Thompson R.E., Hare J.M., et al. Underlying causes and long-term survival in patients with initially unexplained cardiomyopathy. N Engl J Med, 342 (2000), pp. 1077-1084. http://dx.doi.org/10.1056/NEJM200004133421502; Truong J, Yan A.T., Cramarossa G. et. al. Chemotherapy-induced cardiotoxicity: detection, prevention, and management. Can J Cardiol. 2014 Aug; 30 (8): 869-78. doi: 10.1016 / j. cjca. 2014.04.0429; O'Hare M, Sharma A., Murphy K., et. al. Cardio-oncology Part I: chemotherapy and cardiovascular toxicity. Expert Rev Cardiovasc Ther. 2015 Apr 5: 1-8. http://dx.doi.org/10.1586/14779072.2015.1027686]. In patients with lymphoproliferative diseases treated with the anthracycline antibiotic doxorubicin, there is a deterioration in the diastolic function of the left ventricle, a decrease in heart rate, a decrease in cardiac output and cardiac index [Pravdivtseva EV, Poteshkina NG, Svanadze A.M. et al. Prevention of the cardiotoxic effect of the anthracycline antibiotic doxorubicin: the role of an inhibitor of the angiotensin converting enzyme perindopril. Clinician. 2011; 5 (3): 55-60. DOI: 10.17650 / 1818-8338-2011-3-55-60].

The appointment of enalaprilat, verapamil, carvedilol for the purpose of cardioprotection during the course of polychemotherapy with anthracycline antibiotics can prevent the deterioration of diastolic function and cardiac hemodynamics. A number of studies confirm the presence of cardiprotective properties in these drugs [Intracoronary enalaprilat during angioplasty for acute myocardial infarction: alleviation of postischaemic neurohumoral and inflammatory stress? Schaefer U, Kurz T, Bonnemeier H, Dendorfer A, Hartmann F, Schunkert H, Richardt G. J Intern Med. 2007 Feb; 261 (2): 188-200; A critical review of the use of carvedilol in ischemic heart disease. Chen-Scarabelli C, Saravolatz L Jr, Murad Y, Shieh WS, Qureshi W, Di Rezze J, Abrencillo R, Gardin T, Gidwani UK, Saravolatz L, Faggian G, Scarabelli TM. Am J Cardiovasc Drugs. 2012 Dec 1; 12 (6): 391-401. doi: 10.2165 / 11636090-000000000-00000; Egr-1, the potential target of calcium channel blockers in cardioprotection with ischemia / reperfusion injury in rats. Huang Z, Li H, Guo F, Jia Q, Zhang Y, Liu X, Shi G. Cell Physiol Biochem. 2009; 24 (1-2): 17-24. doi: 10.1159 / 000227809. Epub 2009 Jul 1].

Imidazoline receptors are located on the membranes of mitochondria and are actively associated with antioxidant and monoamine oxidase enzymatic systems. In this regard, the correction of mitochondria can significantly offset the manifestations of toxic cardiomyopathy.

In connection with the foregoing, the relevance of the study of the derivative of heterocyclic acid amides (3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid) as a medicine for the correction of chemotherapeutic-induced cardiomyopathy should be noted.

Cardioprotective agents include the drug Mexicor, which is declared for the prevention and treatment of various forms of coronary heart disease (RU No. 2144822, publ. 01/27/2000), which has antianginal activity (RU No. 2168993, publ. 06/20/2001), used to treat acute myocardial infarction (RU No. 2299730, publ. 05.27.2007).

However, the cardioprotective activity of Mexico is insufficiently expressed, which leads to an increase in the duration of treatment and the use of high doses in therapy. Also, the cardioprotective activity of Mexico has serious drawbacks: they act only at the final stage of free radical oxidation, and upon decomposition of free radicals of polyunsaturated fatty acids form toxic products that enhance damage to cell membranes.

The compound (±) -cis-3- (2'-benzimidazolyl) -1,2,2-trimethylcyclopentanecarboxylic acid is known, which exhibits a hypoglycemic and antidiabetogenic effect (RU No. 2205826, publ. 10.06.2003). This compound exhibits high hypoglycemic and antidiabetogenic activity and can be used as a hypoglycemic agent in the manifest forms of type I and type II diabetes mellitus, as well as an antidiabetogenic agent that prevents and / or weakens the development of insulin deficiency in the early stages of the disease.

The objective of the invention is to provide a highly effective compound with cardioprotective activity, which allows to increase the effectiveness of the treatment of cardiovascular diseases.

The problem is achieved by the use of a biologically active compound - 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid, C ₁₆ H ₂₀ N ₂ O ₂ ,

as a cardioprotective agent.

The biologically active compound 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid has a pronounced cardioprotective effect before the reference drugs - enalaprilat, verapamil, carvedilol.

The technical result consists in the use of 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid, C ₁₆ H ₂₀ N ₂ O ₂ , as an agent with pronounced cardioprotective activity, which allows to increase the effectiveness of the treatment of cardiac vascular diseases.

The specified compound, 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid, obtained from 2-aminophenylamide DL-camphoric acid.

METHOD FOR PRODUCING

Example

147 g of DL-camphor anhydride, 0.9 L of toluene and 87 g of phenylenediamine are charged into the flask. Stirring is turned on, the reaction mass is heated to a temperature of 75 ° C and maintained under these conditions for 3 hours. After that, the reaction mass is cooled to room temperature. The precipitate formed is filtered on a Schott filter. The precipitate is thoroughly squeezed and then washed twice on the filter with toluene in 150 ml portions and re-squeezed. The precipitate of 2-aminophenylamide DL-camphoric acid is dried in an oven at a temperature of 70 ° C for 5 hours. Obtain 183 g of 2-aminophenylamide DL-camphoric acid, yield 78%.

Synthesis of 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclo-pentanecarboxylic acid:

183 g of DL-camphoric 2-aminophenylamide, 0.82 L of toluene, 86 ml of dimethylformamide and 87 g of phenylenediamine are charged into the flask. Mixing is turned on and the reaction mass is heated to boiling (110 ° С). The mixture is kept under these conditions until the end of the allocation of water and its collection in the successor nozzle Dean-Stark. About 11 ml of water should be released. After that, the reaction mass is cooled to room temperature.

The precipitate that formed upon standing was filtered off under vacuum on a Schott filter, squeezed thoroughly, and then washed twice with 140 ml portions of toluene on the filter and squeezed again. The precipitate was kept on the filter under vacuum for 1 hour, transferred to a flask and loaded with 370 ml of 70% aqueous ethyl alcohol. Mixing is turned on and the solution is heated to a temperature of 75 ± 5 ° C. The mass is maintained under these conditions for 2 hours, the precipitate of 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclo-pentane-carboxylic acid is filtered off, carefully squeezed out and transferred to a cuvette for drying. The precipitate 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentane-carboxylic acid (techn.) Is dried in an oven at a temperature of 70 ° C for 4 hours. 141 g of product are obtained, yield 82.5%.

For purification, the product is recrystallized twice from ethyl alcohol.

Appearance:

The powder is white or almost white.

Solubility:

It is practically insoluble in water, soluble in 96% alcohol, slightly soluble in chloroform.

Authenticity:

The RG spectrum of the substance, taken in a disk with potassium bromide (1 mg of the substance in 200 mg of potassium bromide) in the region from 4000 to 400 cm ^-1 .

The retention time of the main peak in the chromatogram of the test solution should correspond to the retention time of the main peak in the chromatogram of a standard sample of 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentane-carboxylic acid.

The UV spectrum of a substance should have three maxima at wavelengths of 247 nm, 276 nm and 282 nm.

Melting point from 252 ° C to 258 ° C with decomposition.

Foreign impurities:

Any single unidentified impurity - not more than 0.1%; the amount of impurities is not more than 0.5%.

Mass loss on drying: not more than 1.0%.

Residual Organic Solvents:

Toluene - not more than 0.089%;

Ethanol - not more than 0.5%.

Quantitation:

Not less than 98.0% and not more than 102.0% C ₁₆ H ₂₀ N ₂ O ₂ in terms of anhydrous and free of residual organic solvents.

PHARMACOLOGICAL PROPERTIES OF THE STATED JOINT

The study was performed on isolated hearts of Wistar rats weighing 300 ± 20 g. All rats were divided into 6 experimental groups of 8 animals in each group. The first group is the intraperitoneal injection of saline. The pathology of cardiomyopathies was simulated by administering doxorubicin intraperitoneally at a dose of 20 mg / kg (control group), after 48 hours the parameters of left ventricular contractility were assessed under conditions of a high rhythm of contractions of 480 beats per minute for 15 seconds amid an increase in Ca ²⁺ concentration to 5 mmol in perfusion solution on the heart of rats isolated according to Langendorff. As a criterion for assessing the cardioprotective effect of pharmacological agents, the areas under the curve of the rise in end-diastolic pressure were used. The third group was given 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid at a dose of 50 mg / kg / day orally against the background of the administration of doxorubicin. The comparison drugs used were: enalaprilat (KRKA, Slovenia) 5 mg / kg intraperitoneally every 12 hours, carvedilol (Teva, Israel) orally 1 time per day, verapamil (Alkaloid AO, Macedonia) 5 mg / kg intraperitoneally 1 time per day .

Animals were removed from the experiment after 48 hours. Hearts were removed under zoetil anesthesia (30 mg / kg) and placed in an “ice-cold” (2-4 ° C) Krebs-Henselyait solution of the following composition, mmol: NaCl - 118.5; KCl -4.7; MgSO ₄ / 7H ₂ O - 1.2; KH ₂ PO ₄ - 1.2; CaCl ₂ - 1.5; glucose - 11.1; NaHCO ₃ - 25.0. The pH of the solution throughout the experiment was 7.4. After the cessation of spontaneous contractions, the aorta was isolated and connective tissue was separated. Then the aorta was cannulated and retrograde perfusion of the heart was performed by the Langendorff method in a flow perfusion mode for 20 minutes with a Krebs-Henselyte solution saturated with carbogen (95% O ₂ + 5% CO ₂ ) at 37 ° C and at a pressure of 100 mm Hg and speed perfusion solution 10 ml / min. The contractile function of the heart was recorded using a latex balloon inserted into the cavity of the left ventricle connected to a pressure sensor integrated into the physiological research apparatus MP 150 of Biopac Systems Inc. (California, USA). The can was filled with distilled water, the volume of which was sufficient to create end-diastolic pressure in the left ventricle at the level of 3-5 mm Hg. Using the original AcqKnowledge application of Biopac Systems Inc. (California, USA), all rats were recorded for contractility: left ventricular pressure (LV, mmHg), heart rate (heart rate, beats / min), maximum contraction rate (+ dP / dtmax, mmHg / sec), maximum myocardial relaxation rate (-dP / dtmax, mmHg / sec). To create a high frequency (480 beats per minute), a connector-earth of an electrical stimulator was connected to the cannula, and a connector-plus was connected to the ear of the left atrium. After 20 minutes of perfusion with a high Ca ²⁺ solution (5 mmol / L), the heart was stimulated with electric pulses using an STM 200-1 instrument from Biopac Systems Inc. (California, USA) for 15 seconds.

EXAMPLE OF SPECIFIC PERFORMANCE

In the control group with modeling of doxorubicin cardiomyopathy with the imposition of a submaximal stimulation frequency (480 bpm) and hypercalcium perfusion (5 mmol / l), a “diastole defect” was found, which in digital terms was 8.3 ± 0.3 conv. units, which indicates significant damage and insolvency of calcium pumps of cardiomyocytes. In the intact group, the area was 1.4 ± 0.1 srvc. units, which is 8 times less than in control. The results of functional changes clearly demonstrate the development of pathology against the background of doxorubicin (20 mg / kg) over 48 hours, manifested in the development of a diastole defect with high-frequency cardiostimulation of an isolated rat heart and an increase in the area under the curve for raising the final diastolic pressure. The area under the curve reflects the dynamics under the intraventricular pressure curve. This allows us to assess the degree of myocardial damage and the cardioprotective effects of various studied substances and drugs.

The area of the increase in the final diastolic pressure in the group, which was administered with 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid against the background of the administration of doxorubicin, was the smallest and amounted to 1.5 ± 0.2 conv. units, which indicates the most pronounced cardioprotective effect. Comparison drugs for cardioprotective activity are located in the following sequence: enalaprilat (5 mg / kg), carvedilol (30 mg / kg), verapamil (5 mg / kg) (Table 1).

Note: * - p <0.05 in comparison with the control group, ** - p <0.05 in comparison with the group of intact animals.

Thus, taking into account the importance of changes in the area under the end-diastolic pressure rise curve, the results clearly demonstrate that 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentanecarboxylic acid at a dose of 50 mg / kg / day shows the most pronounced cardioprotective effect compared with enalaprilat, 5 mg / kg; verapamil, 5 mg / kg; carvedilol, 30 mg / kg, which translates into an improvement in contractility in the doxorubicin model of cardiomyopathy.

Claims (3)

The use of 3- (1H-benzimidazol-2-yl) -1,2,2-trimethylcyclopentane-carboxylic acid, C ₁₆ H ₂₀ N ₂ O ₂ ,

as a cardioprotective agent.