MD4215C1 - Inhibitor of human myeloid leukemia based on N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylen)-hydrazinecarbothioamide - Google Patents

Abstract

The invention relates to organic chemistry, namely the synthesis of biologically active compounds in the group of hydrazincarbotioamides with heterocyclic nuclei that can be used for the prophylaxis and treatment of human myeloid leukemia. The essence of the invention is that as an inhibitor of human myeloid leukemia (HL cells) 60) a new compound N- (4- (5- (pyridin-2-yl) -4,5-dihydro-1H-pyrazol-3-yl) phenyl) -2- (pyridin-2-ylmethylene) - is proposed hydrazincarbotioamide of the formula: The claimed compound possesses human myeloid leukemia inhibitor properties.

Description

The invention refers to organic chemistry, namely to the synthesis of biologically active compounds from the group of hydrazinecarbothioamides with heterocyclic nuclei, which can be used for the prophylaxis and treatment of human myeloid leukemia.

Cytarabin (cytarabinum) - 4-amino-1-β-D-arabinofuranosyl-2-(1N)-pyrimidinone (1) with the following structure is widely used in medical practice for the treatment and prophylaxis of leukosis:

(1)

At a concentration of 10-5mol/l this compound inhibits the growth and multiplication of 100% of HL-60 cells of human myeloid leukemia, and at a concentration of 10-6 only 25% [1].

The disadvantage of using this cytostatic preparation consists in causing some side effects, such as: nausea, vomiting, loss of appetite, stomatitis, etc.

The problem that the proposed invention solves is to increase the number of inhibitors of human myeloid leukemia with high biological activity at lower concentrations of the preparation.

The essence of the invention is that a new compound N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazole) is proposed as an inhibitor of human myeloid leukemia (HL-60 cells) -3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide (2) with the following structure:

(2)

The properties and the method of obtaining this compound are not described in the literature.

The technical result of the invention consists in establishing for the mentioned compound (2) the anticancer activity that exceeds 2.5 times at the concentration of 10-6 mol/l and approximately 58 times at the concentration of 10-7 mol/1 the analogous characteristics of 4-amino -1-β-D-arabinofuranosyl-2-(1N)-pyrimidinone (1).

The technical result of the invention is conditioned by the fact that for the first time N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H) is proposed as an inhibitor of HL-60 cells of human myeloid leukemia -pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide (2), which contains a new combination of chemical bonds and already known heterocyclic nuclei.

The proposed compound (2) is obtained in three steps according to the following scheme:

First, by condensing 2-formylpyridine with 3-(4-acetylphenyl)-1,1-dimethylthiourea in basic medium at room temperature, 1,1-dimethyl-3-(4-(3-(pyridyl-2-yl) acryloyl)phenyl)thiourea (3) with a yield of 89%. Next, compound (3) is treated with hydrazine at a molar ratio of 1:2 in pyridine solution for 24 hours, then the temperature is raised to 90...95°C and heated for 2 hours. During this time the intermediate (4) is transformed into N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-hydrazinecarbothioamide (5) in a yield of 77%. The final product N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide (2) is obtained by condensing 2-formylpyridine with compound (5) in ethanol at a temperature of 70...80°C for 2 hours, the yield is 62%. The individuality of the compounds (2, 3, 5) was determined using thin layer chromatography on silufol plates, and their structure was confirmed by elemental and spectral analysis (1H- and 13C-NMR).

Example of obtaining N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazine carbotioamide (2 ) in three stages (a, b, c)

a) Synthesis of 1,1-dimethyl-3-(4-(3-(pyridyl-2-yl)acryloyl)phenyl)thiourea (3). To the solution consisting of 2.22 g (0.01 mol) of 3-(4-acetylphenyl)-1,1-dimethylthiourea and 6 ml of dimethylformamide, 1 g (0.018 mol) of potassium hydroxide dissolved in 4 ml of 95% ethanol is added with stirring, then drop 1.28 g (0.012 mol) of 2-formylpyridine, dissolved in 4 ml of 95% ethanol, at a temperature of 5...10°C. The reactant mixture is left at room temperature for 12 hours, then the solution is filtered of impurities and neutralized to pH~8. 2.78 g (89%) of thiourea (3) with m.p. 182…183°C.

Elemental and spectral NMR analysis. Calculated for C17H17N3OS (3), %: C-65.57, H-5.50, N-13.49, S-10.30. Found, %: C-65.62, H-5.52, N-13.48, S-10.31. 1H-NMR (DMSO-d6), ppm: 3.30 (s, 6H, N(CH3)2), 7.43-8.19 (m, 10H=CH, Ar-H, Py-H), 9.36 (s, 1H, NH ). 13C NMR (DMSO-d6), ppm: 179.70 (C=S), 187.82 (C=O), 181.39 (C=S), 144.47 (C-N), 144.73 (-CH=CH), 126.97 (-CH=CH ), 154.79, 136.82, 131.37, 131.11, 128.84, 40.44, 40.23.

b) Synthesis of N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-hydrazine carbotioamide (5). The mixture formed from 0.62 g (0.002 mol) 1,1-dimethyl-3-(4-(3-(pyridyl-2-yl)acryloyl)phenyl)thiourea (3), 0.22 g (0.0044 mol) hydrazine hydrate and 5 ml of pyridine is left at room temperature for 24 hours, then heated to a temperature of 90...95°C for 2 hours. The crystals obtained are filtered and recrystallized from 98% methanol. 0.48 g (77%) of thioamide (5) with m.p. 199…201°C.

Elemental and spectral NMR analysis. Calculated for C15H16N6S (5), %: C-57.67, H-5.16, N-26.90, S-10.26. Found, %: C-57.62, H-5.20, N-26.91, S-10.25. 1H-NMR (DMSO-d6), ppm: 4.95 (m, 2H, NH2), 7.27-8.58 (m, 8H, =CH, Ar-H, Py-H), 3.42 (m, 2H, CH2), 9.19 (s, NH-CS), 8.53 (s, NH-NH2). 13C NMR (DMSO-d6), ppm: 179.70 (C=S), 162.16 (Py-CH-NH=N), 150.05 (Py-CH(CH2)NH-N=), 139.74 (-C6H4-NH), 64.89 (-CH2-C(-C6H4-)=N), 149.44, 129.28, 125.85, 122.49, 64.89.

c) Synthesis of N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)- hydrazinecarbothioamide (2) . To the solution consisting of 0.62 g (0.002 mol) N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-hydrazinecarbothioamide (5) and 8 ml dimethylformamide, 0.22 g (0.002 mol) of 2-formylpyridine, dissolved in 1 ml of 95% ethanol, is added with stirring. The reactant mixture is heated to a temperature of 70...80°C for 2 hours, then diluted with a little water and cooled. The crystalline product is filtered and recrystallized from 95% ethanol. 0.48 g (63%) of final product (2) with m.p. 140...142°C.

Elemental and spectral NMR analysis. Calculated for C21H19N7S (2), %: C-62.82, H-4.77, N-24.42, S-7.99. Found, %: C-62.85, H-4.75, N-24.41, S-7.98. 1H-NMR (DMSO-d6), ppm: 6.10-8.67 (m, 13H, =CH, Ar-H, Py-H), 3.33 (m, 2H, CH2), 10.42 (s, NH-CS), 12.22 (s, NH-N=CH). I3C NMR (DMSO-d6), ppm: 176.66 (C=S), 158.18 (Py-CH-NH=N), 152.16 (-NH-N=CH-Py), 152.07 (Py-CH(CH2)NH- N=), 139.30 (-C6H4-NH), 64.95 (-CH2-C(-C6H4-)=N), 129.28, 143.98, 138.89, 127.79.

The process for obtaining N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazine carbotioamide (2 ) includes three stages, but is simple in execution and uses accessible initial substances. The claimed product is stable, slightly soluble in water, better soluble in ethanol, well soluble in dimethylformamide and dimethylsulfoxide, practically insoluble in hexane or ether.

The experimental data obtained regarding the study of the anticancer properties of compound (2) show that it inhibits the growth of HL-60 cells of human myeloid leukemia within the concentration limits of 10-5...10-7mol/l. At the concentration of 10-6mol/l it inhibits 62.6%, and at the concentration of 10-7 -58% of HL-60 cells of human myeloid leukemia. The data included in the table indicate that compound (2) claimed for its anticancer activity exceeds 2.5 times at the concentration of 10-6mol/l and 58 times at the concentration of 10-7mol/l the analogous characteristics for the closest solution [1].

Table

The fraction of inhibited cells of human myeloid leukemia HL-60, %

Compound Concentration, mol/l 10-5 10-6 10-7 Cytarabine (closest solution) 100 25 0 N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazole) -3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide (claimed compound) - 62.6 58

The biological properties of the claimed compound are of interest for medicine from the point of view of expanding the arsenal of inhibitors of human myeloid leukemia at lower concentrations of the preparation and reducing the negative effects: nausea, vomiting, loss of appetite, stomatitis, leukopenia, anemia and phlebitis at the sites of infection, characteristics for the closest solution.

N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide (2) belongs to the class of compounds pyrazoles, which have been found to have antibacterial, antifungal, local anesthetic activities, including sedative and antituberculosis activities (Mohamed Ashraf Ali, Mohammad Shaharyar*, Anees Ahamed Siddiqui, Synthesis, structural activity relationship and anti-tubercular activity of novel pyrazoline derivatives Faculty of Pharmacy , Jamia Hamdard University, Department of Pharmaceutical Chemistry, Hamdard Nagar, New Delhi 110062, India, Received 22 February 2006; received in revised form 22 July 2006; accepted 11 August 2006, Available online 26 September 2006, European Journal of Medicinal Chemistry, 42 (2007), 268-276, Elsevier). Some 1,3,5-trisubstituted pyrazoline derivatives have anticonvulsant and antidepressant properties (Ozan Ruhoglu, Zuhal Özdemira, Ünsal Çalis a, Bülent Gümüselb, and Abdullah Altan Bilgina, Synthesis of and Pharmacological Studies on The Antidepressant and Anticonvulsant Activities of Some 1,3 ,5-Trisubstituted Pyrazolines Arzneim.-Forsch./Drug Res. 55, No. 8, 431-436 (2005), Editio Cantor Verlag, Aulendorf (Germany)).

1. Mashkovsky M. D. Medicines. Moscow, Novaya volna, 2008, p. 1206

Claims (2)

1. N-(4-(5-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)phenyl)-2-(pyridin-2-ylmethylene)-hydrazinecarbothioamide with the formula: . 2. Compound according to claim 1, characterized in that it possesses inhibitory properties of human myeloid leukemia.