Preparation method and use of benzamide derivatives containing aryloxy bipyridylmethylthiourea units

Abstract

The present invention relates to the preparation and use of benzamide derivatives containing aryloxybipyridylmethylthiourea units. These derivatives are obtained by condensing a substituted o-phenylenediamine with benzoic acid containing aryloxybipyridylmethylthiourea units. These benzamide derivatives exhibit a significant inhibitory effect on tumor cells and can be used to prepare anti-tumor cell drugs. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a preparation method and application of a benzamide derivative containing an aryloxy bipyridylmethylthiourea unit. Background Art

[0002] Malignant tumors are one of the major diseases that seriously affect human health. In recent years, the incidence of cancer has shown an upward trend. Therefore, the development of effective anticancer drugs and treatments is a hot research area in the medical community.

[0003] Pyridine compounds are important nitrogen-containing six-membered heterocycles. Many compounds containing pyridine heterocyclic units exhibit excellent inhibitory effects on tumor cells. Benzamide derivatives, such as entinostat, also exhibit excellent inhibitory effects on tumor cells.

[0004] Therefore, in order to continue to develop drugs with good anti-tumor activity from benzamide derivatives, the substituted pyridine skeleton is organically linked to the benzamide unit. The present invention discloses a class of benzamide derivatives containing aryloxy bipyridylmethylthiourea units with medicinal value. Summary of the Invention

[0005] The purpose of the present invention is to provide a class of benzamide derivatives containing aryloxy bipyridylmethylthiourea units that exhibit good inhibitory effects on human gastric cancer cells HGC-27 and human colon cancer cells HCT116.

[0006] Another object of the present invention is to provide a method for preparing the above compound.

[0007] Another object of the present invention is to provide the use of the above compound in the preparation of anti-tumor cell drugs.

[0008] In order to solve the above technical problems, the present invention provides a benzamide derivative containing an aryloxy bipyridylmethylthiourea unit, which has a general structure of formula I,

[0009]

[0010] Preferably, the benzamide derivative containing an aryloxybipyridylmethylthiourea unit has the following structure:

[0011]

[0012] The present invention provides a method for preparing the above-mentioned benzamide derivative containing an aryloxybipyridylmethylthiourea unit, which is characterized by comprising the following steps:

[0013] The intermediate III is dissolved in an organic solvent, and the intermediate II is added. After a period of reaction, the reaction is stopped. After removing the solvent, the crude product is purified by silica gel column chromatography to obtain the target compound I.

[0014]

[0015] Preferably, the preparation method of the benzamide derivative containing an aryloxybipyridylmethylthiourea unit comprises the following reaction steps:

[0016]

[0017]

[0018] Compound I has a good inhibitory effect on tumor cells, such as human gastric cancer cells HGC-27 and human colon cancer cells HCT116.

[0019] The benzamide derivative containing an aryloxy bipyridylmethylthiourea unit disclosed in the present invention exhibits good inhibitory activity against human gastric cancer cells HGC-27 and human colon cancer cells HCT116, and can therefore be used to prepare anti-tumor cell drugs. DETAILED DESCRIPTION

[0020] In order to facilitate a further understanding of the present invention, the following examples are provided to illustrate it in more detail. These examples are for narration only and are not intended to limit the scope or implementation principles of the present invention.

[0021] Example 1:

[0022]

[0023] Dissolve 3 mmol of intermediate IIIa and 4 mmol of benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) in 30 mL of DMF. Add 6 mmol of 4-dimethylaminopyridine (DMAP) and 3 mmol of intermediate IIa at room temperature. Stir at room temperature for 12 hours. After removing the solvent, the crude product is purified by silica gel column chromatography to obtain the target compound Ia. 1HNMR (400MHz, DMSO-d6): δ9.92 (s, 1H, NH), 9.57 (s, 1H, NH), 8.43 (s, 1H, NH), 8.08 (d, J=2.0Hz, 1H, Py-H), 7.90~7.94 (m, 3H, Ar-H and Py-H), 7.78 (d, J=2.4Hz, 1H, Ar-H), 7.34~7.60 (m, 4H, Ar-H and Py-H), 7.14~7.16 (m, 2H, Ar-H), 6.80 (d, J=2.4Hz, 1H, Ar-H), 6.58~6.60 (m, 1H, Ar-H), 5.24 (s, 2H, NH2), 4.71 (d, J=5.2Hz, 2H, CH2).

[0024] Example 2:

[0025]

[0026] Dissolve 4 mmol of intermediate IIIb and 4 mmol of benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) in 35 mL of DMSO. Add 10 mmol of triethylamine and 5 mmol of intermediate IIb at room temperature. After addition, control the temperature at 40°C for 16 hours. After removing the solvent, the crude product is purified by silica gel column chromatography to obtain the target compound Ib. 1 H NMR (400MHz, DMSO-d6): δ9.90 (s, 1H, NH), 9.53 (s, 1H, NH), 8.41 (s, 1H, NH), 8.13 (d, J=1.6Hz, 1H, Py-H), 7.86~7.95 (m, 3H, Ar-H and Py-H), 7.59 (d, J=7.6Hz, 2H, Ar-H), 7.09~7.27 (m, 5H, Ar-Hand Py-H), 7.04 (d, J=8.4Hz, 1H, Ar-H), 6.53~6.56 (m, 1H, Ar-H), 6.33~6.38 (m, 1H, Ar-H), 5.22 (s, 2H, NH2), 4.71 (d, J=5.2Hz, 2H, CH2).

[0027] Example 3:

[0028]

[0029] Dissolve 5 mmol of intermediate IIIc and 6 mmol of benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) in 40 mL of DMF. Add 20 mmol of triethylamine and 6 mmol of intermediate IIc at room temperature. Stir at room temperature for 10 hours. After removing the solvent, the crude product is purified by silica gel column chromatography to obtain the target compound Ic. 1 H NMR (400MHz, DMSO-d6): δ9.93 (s, 1H, NH), 9.61 (s, 1H, NH), 8.43 (s, 1H, NH), 8.15 (d, J=2.0Hz, 1H, Py-H), 7.89~7.96 (m, 3H, Ar-H and Py-H), 7.59 (d, J=8.8Hz, 2H, Ar-H), 7.34~7.52 (m, 2H, Ar-Hand Py-H), 6.95~7.16 (m, 4H, Ar-H), 6.77~6.79 (m, 1H, Ar-H), 6.58~6.62 (m, 1H, Ar-H), 4.90 (s, 2H, NH2), 4.73 (d, J=5.2Hz, 2H, CH2).

[0030] Example 4:

[0031]

[0032] Dissolve 6 mmol of intermediate IIId, 6 mmol of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), and 6 mmol of 1-hydroxybenzotriazole (HOBt) in 25 mL of DMF. Add 7 mmol of intermediate IIc at room temperature. Stir at room temperature for 11 hours. After removal of the solvent, the crude product is purified by silica gel column chromatography to yield the desired compound Id. 1 HNMR (400MHz, DMSO-d6): δ9.93 (s, 1H, NH), 9.60 (s, 1H, NH), 8.44 (s, 1H, NH), 8.16 (s, 1H, Py-H), 7.92~7.96 (m, 3H, Ar-H and Py-H), 7.45~7.68 (m, 6H, Ar-H and Py-H), 7.13~7.17(m, 2H, Ar-H), 6.95~6.99(m, 1H, Ar-H), 6.78(d, J=8.0Hz, 1H , Ar-H), 6.58~6.62 (m, 1H, Ar-H), 4.90 (s, 2H, NH2), 4.75 (d, J=4.8Hz, 2H, CH2).

[0033] Example 5:

[0034]

[0035] 6 mmol of intermediate IIIe and 5 mmol of benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) were dissolved in 35 mL of DMSO. 18 mmol of 4-dimethylaminopyridine (DMAP) and 5 mmol of intermediate IIc were added at room temperature. After addition, the reaction was continued at room temperature for 20 hours. After removal of the solvent, the crude product was purified by silica gel column chromatography to obtain the target compound Ie. 1 HNMR (400MHz, DMSO-d6): δ9.89 (s, 1H, NH), 9.60 (s, 1H, NH), 8.40 (s, 1H, NH), 8.05 (s, 1H, Py-H), 7.81~7.95 (m, 3H, Ar-H and Py-H), 7.59 (d, J=8.4Hz, 2H, Ar-H), 7.10~7.24 (m, 4H, Ar-H and Py-H), 6.94~6.99 (m, 3H, Ar-H), 6.78 (d, J=7.6Hz, 1H, Ar-H), 6.58~6.62 (m, 1H, Ar-H), 4.89 (s, 2H, NH2), 4.69 (d, J=4.4Hz, 2H, CH2), 3.69 (s, 3H, OCH3).

[0036] Example 6:

[0037] Screening of sample activity against tumor cells

[0038] The in vitro antitumor activity of the compounds was determined using the MTT colorimetric assay. The test subjects were human gastric cancer cells HGC-27 and human colon cancer cells HCT116. 5-Fluorouracil (5-FU) was selected as the positive control drug. 4×10 3 A cell suspension of 10 cells / mL was inoculated into a 96-well plate and cultured in a 37°C, 5% CO2 incubator for 24 hours. Then, the test solution (10 μL) of the sample to be tested was added to the test well, 5 parallel wells were set for each concentration, and an equal amount of dimethyl sulfoxide was used as a blank control. After 72 hours of culture in a 5% CO2 incubator, the supernatant was discarded, and 20 μL of MTT (2 mg / mL in PBS) was added to each well. After 4 hours of culture, the culture medium was aspirated and discarded, and 150 μL of dimethyl sulfoxide was added to each well. The blue-purple precipitate was dissolved by shaking on a vibrator for 10 minutes. The OD value was measured at a wavelength of 490 nm using an enzyme marker to calculate the cell inhibition rate. Cell inhibition rate = (OD value of the negative control group - OD value of the test substance group) / OD value of the negative control group × 100%, and the IC value of the compound was calculated by the probability unit weighted regression method. 50 value.

[0039] Table 1. Antitumor activity data of Ia-Ie (IC 50 , μM)

[0040] Compound HGC-27 HCT116 Ia 0.40 1.33 Ib 1.34 1.91 Ic 2.11 3.29 Id 1.42 2.12 Ie 0.33 2.21 5-FU 34.10 19.60

[0041] As shown in Table 1, compounds Ia-Ie exhibited significant inhibitory activity against both human gastric cancer cells HGC-27 and human colon cancer cells HCT116. These experimental results demonstrate that the compounds synthesized by organically linking a substituted pyridine structure with a benzamide backbone exhibited excellent antitumor activity against both human gastric cancer cells HGC-27 and human colon cancer cells HCT116.

[0042] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above examples. The above examples and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications are possible without departing from the spirit and scope of the present invention. Such changes and modifications are intended to fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A benzamide derivative (I) containing an aryloxybipyridylmethylthiourea unit, characterized in that The structure is:

2. The method for preparing the benzamide derivative (I) containing an aryloxybipyridylmethylthiourea unit according to claim 1, characterized in that Here’s how:

3. Use of the benzamide derivative (I) containing an aryloxybipyridylmethylthiourea unit as claimed in claim 1 in the preparation of an anti-tumor cell drug, characterized in that: The tumor cells are human gastric cancer cells HGC-27 or human colon cancer cells HCT116.

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