RU2456284C1 - 2-aminopropylmorpholino-5-aryl-6h-1,3,4-thiadiazines, dihydrobromides and 2-aminopropylmorpholino-4-arylthiazoles, hydrobromides showing antiaggregant action - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to 2-aminopropylmorpholino-5-aryl-6H-1,3,4-thiadiazines, dihydrobromides (of general formula I) and 2-aminopropylmorpholino-4-arylthiazoles, hydrobromides, (of general formula II) which show antiaggregant activity.

wherein R = H; F; CI; Br; OH.

EFFECT: given compounds may be used for preparing cardiologic drugs and enable better treatment of various cardiovascular diseases, including myocardial infarction and thrombotic apoplexy.

1 cl, 2 tbl

Description

The invention relates to the field of biologically active compounds, for the development of new derivatives of the class 5-aryl-1,3,4-thiadiazine and 4-arylthiazole with an aminopropylmorpholino substituent at position 2 of both the 1,3,4-thiadiazine and thiazole rings and possessing antiaggregant action.

Thrombosis plays a leading role in the occurrence of cardiovascular disease. The most important role in the formation of blood clots is platelet aggregation. Due to the interaction of agonists (ADP, arachidonic acid) and platelet receptors, platelet activation occurs. Activated platelets cross-interact with each other, which leads to the formation of a platelet plug. Pharmacological modulation of platelet aggregation activity is one of the leading directions in the correction of hemostatic disorders, the solution of which will significantly improve the treatment of myocardial infarction, thrombotic stroke (these diseases are the leading cause of death in developed countries).

Currently, the world is actively developing methods for targeted chemical synthesis of new generation antiplatelet agents with high efficiency and selectivity. In recent years, an effective foreign platelet aggregation inhibitor from the group of thienopiperidine derivatives - “clopidogrel” has been developed and introduced into medical practice [1]. The therapeutic effect of the drug develops only on the second day and reaches a maximum only on the 4-7th day of treatment. It has a number of contraindications, as may cause internal bleeding. In addition, complications such as diarrhea, rash, and leukopenia are common in its use. To obtain the necessary faster antiplatelet effect, it is often used with aspirin, but even in this case it is not always possible to exclude internal bleeding.

In the ranks of 1,3,4-thiadiazines, we have previously found substances that possess a unique combined anticoagulant and antiplatelet effect and are effective for intravenous administration [2]. This forces us to expand the search for new compounds in this class in order to identify substances of leaders for the behavior of preclinical trials.

The technical result of this invention is the creation of new chemical compounds of class 5-aryl-1,3,4-thiadiazine and class 4-arylthiazole having the same aminopropylmorpholino substituent at position-2 of the hetero ring and having good solubility in water.

The specified technical result is achieved by the fact that according to the invention, new 2-aminopropylmorpholino-5-aryl-6H-1,3,4-thiadiazines, dihydrobromides (general formula 1) and 2-aminopropylmorpholino-4-arylthiazoles, hydrobromides (general formula 2) are synthesized with high antiplatelet activity in vitro tests on human platelets

where R = H; F; Cl; Br; OH.

These compounds can be used to create domestic drugs for cardiology and will significantly improve the treatment of various cardiovascular diseases, including myocardial infarction and thrombotic stroke.

Close in structure (prototypes) with respect to the claimed compounds are, respectively, 2-morpholino-5-phenyl-6H-1,3,4-thiadiazine, total hydrobromide (formula III) and 2-morpholino-4-phenylthiazole (formula IV)

The claimed compounds of formulas I and II differ from the prototype, respectively (formulas III and IV) in that at position-2 of 5-aryl-1,3,4-thiadiazine and 4-arylthiazole there is a residue of 2-aminopropylmorpholine, and in the case of 2-aminopropylmorpholino -4-arylthiazole (II) - by the presence of substituents on phenyl in position-4 of the thiazole ring.

For 2-morpholino-5-aryl-6H-1,3,4-thiadiazines (formula III), the synthesis and antiplatelet activity detected for them are protected by a patent [2].

2-Morpholino-4-phenylthiazole (IV) was obtained during the photochemical reaction from 2-morpholino-5-phenyl-6H-1,3,4-thiadiazine (III) [3], which indicates a high lability of 1,3,4 -thiadiazine ring and the likely possibility of its metabolism in the thiazole cycle and in the body. However, it was not possible to test the biological activity of 2-morpholino-4-phenylthiazole (IV) due to its poor solubility in water. All salts of this compound were unstable and, when dissolved in water, were hydrolyzed to the base, 2-morpholino-4-phenylthiazole.

This patent application made it possible to compare the antiplatelet activity of 1,3,4-thiadiazines and thiazoles due to the synthesis of new compounds with the same and more basic substituent, 2-aminopropylmorpholine. An advantage of the claimed compounds of general formulas I and II, in comparison with their corresponding prototypes III and IV, is the stability of their salt structures (dihydrobromides for I and hydrobromides for II) and, as a result, their good solubility in water. This is very important for the penetration of drugs into the cell and achieve the necessary concentration of the drug.

The biological activity of compounds I and II with the ADP aggregate was compared in an experiment with the effective foreign drug “clopidogrel” [1] (formula V)

1. Examples of synthesis and physico-chemical characteristics of the claimed compounds

The synthesis of 2-aminopropylmorpholino-5-aryl-6H-1,3,4-thiadiazines, dihydrobromides (I) is based on cyclocondensation of various α-bromoacetophenones and 4- [3- (4-morpholino) propyl] -3-thiosemicarbazide.

The synthesis of 2-aminopropylmorpholino-4-phenylthiazoles, hydrobromides (II) is based on cyclocondensation of α-bromoacetophenones and 4- (3-aminopropyl) morpholinyl thiourea (Scheme 1).

Scheme 1

2-aminopropylmorpholino-5-phenyl-6H-1,3,4-thiadiazine, dihydrobromide (1).

To 1.0 g (5 mmol) of α-bromoacetophenone in 40 ml of absolute ethanol was added 1.09 g (5 mmol) of 4- [3- (4-morpholino) propyl] -3-thiosemicarbazide and 0.5 ml of concentrated HBr, boiled for 40 minutes After hot filtration and cooling, 70 ml of dry diethyl ether was added, kept for 2 hours in ice, the precipitated colorless crystalline precipitate was filtered off, dried and crystallized from isopropanol. Yield 1.3 g (54%). T. pl. 223-224 ° C. R _f = 0.30 (eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 40.06; H 5.35; N, 11.70. C ₁₆ H ₂₄ Br ₂ N ₄ OS. Calculated,%: C 40.0; H 5.0; N, 11.67. ¹ H NMR (DMSO-d ₆ ), δ, ppm: 2.20 (t, 2H, CH ₂ ), 3.31 (s, 2H, CH ₂ ), 3.34 (m, 4H, morpholino ), 3.67 (s, 2H, CH ₂ ), 3.96 (m, 4H, morpholino), 4.32 (s, 2H, CH ₂ S), 7.52-7.54 and 7.93- 7.95 (m, 5H, C ₆ H ₅ ), 10.54 (br s, 2H, NH), 12.34 (br s, 1H, NH).

2-aminopropylmorpholino-5- (4'-chlorophenyl) -6H-1,3,4-thiadiazine, dihydrobromide (2)

To 2.34 g (10 mmol) of α-bromo-4'-chloroacetophenone in 90 ml of absolute ethanol was added 2.18 g (10 mmol) of 4- [3- (4-morpholino) propyl] -3-thiosemicarbazide and 1, 2 ml of concentrated HBr. Boiled for 45 minutes, cooled and 170 ml of dry diethyl ether was added. After 40 minutes in ice, the slightly yellow precipitate formed was filtered off and crystallized from a mixture of isopropanol: absolute ethanol 5: 1. Yield 2.9 g (56%). T. pl. 172-173 ° C. R _f = 0.28 ((eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 37.42; H 4.47; N 10.74. C ₁₆ H ₂₃ Br ₂ ClN ₄ OS. Calculated,%: C 37.32; H 4.50; N 10.88. NMR ¹ H (DMSO-d ₆ ), δ, ppm: 2.20 (t, 2H, CH ₂ ), 3.17 (t, 2H, CH ₂ ), 3.27 (m, 4H, morpholino), 3.45 (s, 2H, CH ₂ ), 3.96 (m, 4H, morpholino), 4, 31 (s, 2H, CH ₂ S), 7.53 (d, 2H, C ₆ H ₄ ), 7.96 (d, 2H, C ₆ H ₄ ), 10.62 (br s, 2H, NH), 13.67 (br s, 1H, NH).

2-aminopropylmorpholino-5- (4'-hydroxyphenyl) -6H-1,3,4-thiadiazine, dihydrobromide (3)

To 2.15 g (10 mmol) of α-bromo-4'-hydroxyacetophenone in 80 ml of absolute ethanol was added 2.18 g (10 mmol) of 4- [3- (4-morpholino) propyl] -3-thiosemicarbazide and 1, 0 ml of concentrated HBr was boiled in a water bath for 40 minutes. After hot filtration and cooling, 150 ml of dry diethyl ether were added and kept in ice for 1 h. The precipitated colorless crystalline precipitate was filtered off and crystallized from a mixture of absolute ethanol: dry diethyl ether 2: 1. Yield 2.54 g (51%). Mp 208-209 ° C. R _f = 0.29 ((eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 38.65; H 4.73; N 11.20. C ₁₆ H ₂₄ Br ₂ N ₄ O ₂ S. Calculated,%: C 38.71; H 4.84; N 11.29. NMR ¹ H (DMSO-d ₆ ), δ, ppm: 2.21 (t, 2H, CH ₂ ), 3.18 (m, 4H, morpholino), 3.42 (t, 2H, CH ₂ ), 3.51 (t, 2H, CH ₂ ), 3.95 (m, 4H, morpholino), 4, 23 (s, 2H, CH ₂ S), 6.87 (d, 2H, C ₆ H ₄ ) and 7.77 (d, 2H, C ₆ H ₄ ), 10.04 (br.s.c., 1H, NH), 10.05 (br s, 2H, NH), 13.51 (br s, 1H, OH).

2-aminopropylmorpholino-5- (4'-fluorophenyl) -6H-1,3,4-thiadiazine, dihydrobromide (4)

To 2.18 g (10 mmol) of α-bromo-4-fluoroacetophenone in 70 ml of absolute ethanol was added 2.18 g (10 mmol) of 4- [3- (4-morpholino) propyl] -3-thiosemicarbazide and 1 ml of concentrated HBr, boiled in a water bath for 45 minutes, was filtered off, cooled and 150 ml of dry diethyl ether was added. The reaction mixture was kept on ice for 1.5 hours, the precipitated colorless precipitate was filtered off, and crystallized from isopropanol. Yield 2.58 g (54%). T. pl. 210-211 ° C. R _f = 0.29 (eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 38.45; H 4.67; N 11, 36. C ₁₆ H ₂₃ Br ₂ FN ₄ OS. Calculated,%: C 38, 55; H 4.62; N, 11.24. ¹ H NMR (DMSO-d ₆ ), δ, ppm: 2.20 (t, 2H, CH ₂ ), 3.18 (t, 2H, CH ₂ ), 3.27 (m, 4H, morpholino ), 3.67 (s, 2H, CH ₂ ), 3.96 (m, 4H, morpholino), 4.32 (s, 2H, CH ₂ S), 7.29 (t, 2H, C ₆ H ₄ ), 8.02 (dd, 2H, C ₆ H ₄ ), 10, 62 (br.s, 2H, NH), 13.70 (br.s, 1H, NH).

2-aminopropylmorpholino-4-phenylthiazole, hydrobromide (5)

To 1 g (5 mmol) of α-bromoacetophenone in 50 ml of absolute ethanol was added 1 g (5 mmol) of 4- (3-aminopropyl) morpholinyl thiourea in 50 ml of absolute ethanol and heated for 3.5 hours in a water bath, then cooled for 2 hours. The precipitated colorless crystalline precipitate was filtered off, dried and crystallized from isopropanol with the addition of activated carbon. Yield 1.2 g (63%). R _f = 0.46 (eluent, n-butanol: acetic acid: water - 4: 1: 5). T. pl. 167-168 ° C. Found,%: C 50.05; H 6.09; N, 11.0. C ₁₆ H ₂₂ BrN ₃ OS. Calculated,%: C 50.0; H 5.72; N, 10.93. ¹ H NMR (DMSO-d ₆ ), δ, ppm: 2.09 (t, 2H, CH ₂ ), 3.23 (m, 4H, morpholino), 3.25 (t, 2H, CH ₂ ), 3.43 (t, 2H, CH ₂ ), 3.90 (m, 4H, morpholino), 6.86 (s, 1H thiazole), 7.22-7.34 and 7.78-7.81 (m, 5H, C ₆ H ₅ ), 7.69 (br s, 1H, NH), 10.45 (br s, 1H, NH).

2-aminopropylmorpholino-4- (4'-chlorophenyl) thiazole, hydrobromide (6)

To 1.17 g (5 mmol) of α-bromo-4'-chloroacetophenone in 40 ml of absolute ethanol was added 1.02 g (5 mmol) of 4- (3-aminopropyl) morpholinyl thiourea in 30 ml of absolute ethanol, boiled for 4.5 hours in a water bath, then the solvent was distilled off in vacuo. To the oily residue, 30 ml of dry diethyl ether was added and kept under cooling for 1.5 hours. The colorless precipitate formed was filtered off, dried and crystallized from isopropanol. Yield 1.1 g (53%). T. pl. 169-170 ° C. R _f = 0.54 (eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 45.74; H, 5.12; N, 10.11. C ₁₆ H ₂₁ BrClN ₃ OS. Calculated,%: C 45.90; H 5.06; N, 10.03. ¹ H NMR (DMSO-d ₆ ), δ, ppm: 2.10 (t, 2H, CH ₂ ), 3.11 (s, 2H, CH ₂ ), 3.24 (t, 2H, CH ₂ ) 3.42 (m, 4H, morpholino), 3.90 (m, 4H, morpholino), 6.95 (s, 1H thiazole), 7.31 (dd 2H, C ₆ H ₄ ), 7 72 (br s, 1H, NH), 7.80 (dd, 2H, C ₆ H ₄ ), 10.45 (br s, 1H, NH).

2-aminopropylmorpholino-4- (4'-bromophenyl) thiazole, hydrobromide (7)

To 1.4 g (5 mmol) of α-bromo-4'-bromoacetophenone in 50 ml of absolute ethanol was added 1.02 g (5 mmol) of 4- (3-aminopropyl) morpholinyl thiourea, boiled for 5 hours in a water bath. To the cooled solution was added 100 ml of dry diethyl ether and kept under ice cooling for 1 h. The resulting colorless precipitate was filtered off, dried and crystallized from isopropanol. Yield 1.5 g (65%). T. pl. 166-167 ° C. R _f = 0.56 (eluent, n-butanol: acetic acid: water 4: 1: 5). Found,%: C 41.38; H 4.58; N, 9.12. C ₁₆ H ₂₁ Br ₂ N ₃ OS. Calculated,%: C 41.47; H 4.54; N, 9.07. ¹ H NMR (DMSO-d ₆ ), δ, ppm: 2.13 (t, 2H, CH ₂ ), 3.28 (t, 2H, CH ₂ ), 3.47 (s, 2H, CH ₂ ), 3.53 (m, 4H, morpholino), 3.90 (m, 4H, morpholino), 7.13 (s, 1H thiazole), 7.57 (dd, 2H, C ₆ H ₄ ), 7.75 (dd, 2H, CH ₂ ), 13.48 (br s, 3H, NH).

2. The study of antiplatelet activity in vitro

Materials and methods

Working solutions of the studied compounds were prepared by diluting them in distilled water at room temperature or by heating to 50-70 ° C until the required concentration was reached.

Experiments to study the anti-aggregation ability of the compounds were performed using venous blood from healthy donors (n = 50), which was obtained by puncture of the cubital vein and stabilized with a 3.8% solution of sodium citrate in a ratio of 9: 1. To prepare platelet-rich plasma, blood was centrifuged for 10 minutes at 1000 rpm, after which the upper plasma layer was transferred to another tube, and the residue was re-centrifuged for 20 minutes at 3000 rpm to obtain platelet-poor plasma. Platelet aggregation was studied on an aggregometer from Chrono-Log Corporation (USA) according to the method of G. Born [4]. For this purpose, 250 μl of platelet-rich plasma was placed in the cuvette of the device. The same volume of platelet-free plasma was used as optical control. The degree of aggregation was judged by the maximum optical density drop after the end of the reaction (A _max ) compared to the initial value. The experiments were carried out as follows: in a cuvette containing 250 μl of platelet-rich plasma, 28 μl of the studied drug was added and the resulting mixture was incubated for 5 min at 37 ° C and platelet aggregation was induced. In control experiments, only a proaggregant was added to the platelet-rich plasma. Arachidonic acid (AK) at a final concentration of 1 × 10 ^-3 M and ADP (Boehringer Mannheim, Germany) at a final concentration of 1 × 10 ^-5 M, arachidonic acid (AK), free of oxidation products, were used as proaggregants in the work. provided by the staff of the laboratory of oxilipids of the Institute of bioorganic chemistry named after M.M.Shemyakin and Yu.A. Ovchinnikov of the Russian Academy of Sciences.

The results of the study of the antiplatelet properties of the compounds are shown in table 1 and table 2.

Industrial applicability

These compounds can be used as potential pharmacologically active substances in the production of drugs of the cardiovascular spectrum of action, allowing to regulate hemostasis and inhibit thrombus formation.

Sources of scientific and technical information

1. Chua D, Ignaszewski A. Clopidogrel in acute coronary syndromes. BMJ. 2009 Apr 14; 338: b1180.

2. RF patent No. 2259371 (2005). Chupakhin OH, Sidorova L.P., Perova N.M., Charushin V.N., Rusinov V.L. and others. "Substituted 5R ₁ , 6R ₂ -1,3,4-thiadiazin-2-amines and pharmaceutical compositions containing them as pharmacologically active agents with anticoagulant and antiplatelet effect."

3. Perova N.M., Egorova L.G., Sidorova L.P., Novikova A.P., Chupakhin O.N. // Transformation of 2-cycloalkylimino-5-phenyl-6H-1,3,4-thiadiazines under UV irradiation // Chemistry of heterocyclic compounds. 1993, No. 4. S.565-566.

4. Born G. Aggregation of blood platelets by adenosine diphosphate and its reversal Nature (London), 194, 927-929 (1962).

Claims (1)

2-aminopropylmorpholino-5-aryl-6H-1,3,4-thiadiazines, dihydrobromides and 2-aminopropylmorpholino-4-arylthiazoles, hydrobromides with antiaggregant action

where R = H; F; Cl; Br; IT.