WO2023130540A1

WO2023130540A1 - Pyrazol benzamide compound, preparation method therefor, and application thereof

Info

Publication number: WO2023130540A1
Application number: PCT/CN2022/077623
Authority: WO
Inventors: 李环球; 汪维鹏; 吕依昕; 朱一凡; 魏若男; 郭绪芹
Original assignee: 苏州大学
Priority date: 2022-01-06
Filing date: 2022-02-24
Publication date: 2023-07-13
Also published as: CN114394934A; CN114394934B

Abstract

Provided are a pyrazol benzamide compound represented by formula (I), a preparation method therefor and an application thereof, wherein R 1 and R 2 are independently selected from hydrogen, alkyl, amino, nitro, halogen, alkoxy, trifluoromethyl, cyano, cycloalkyl, heterocyclyl, -COR 3, -COOR 4, -SO 2 R 5, and -NR 6 R 7. Hydrogen cannot be selected for R 1 and R 2 at the same time. Compared with the prior art, the pyrazol benzamide compound has good anti-tumor activity and can be used for preparing drugs treating tumor diseases.

Description

Pyrazole benzamide compound and its preparation method and application

This application claims the priority of the Chinese patent application submitted to the China Patent Office on January 06, 2022, with the application number 202210012535.5, and the invention name is "pyrazole benzamide compounds and their preparation methods and applications", all of which are passed References are incorporated in this application.

technical field

The invention belongs to the technical field of medicinal chemistry, and in particular relates to pyrazole benzamide compounds and their preparation methods and applications.

Background technique

Malignant tumors are the second deadliest disease in the world, while lung cancer, prostate cancer, colorectal cancer, and breast cancer rank first. The morbidity and mortality of cancer have continued to rise in the past decade. There are many conventional treatment methods such as surgery, chemotherapy, radiotherapy and so on. However, most traditional treatment methods cannot achieve a curative effect, especially for patients with advanced malignant tumors, and the mortality rate remains high. Small-molecule targeted anti-tumor drugs have gradually become a hot topic in the development of innovative drugs at home and abroad because of their high efficacy, low toxicity and side effects, and high specificity. Different from traditional drugs, small molecule drugs can specifically bind to human targets and effectively inhibit primary tumors. They have low toxicity, high bioavailability, wide range of biological activities, and good biocompatibility and curative effect.

At present, many anti-cancer drugs on the market generally have the characteristics of high toxicity, strong side effects, and high cost. However, small molecule target anti-cancer drugs have gradually become the research and development of anti-tumor therapeutic drugs due to their excellent properties such as low toxicity, high efficiency, and high selectivity. Finding more ideal targets and related inhibitors is a key problem to be solved in tumor therapy.

Pyrazole benzamides are important pharmacophores in modern drug discovery. A number of outstanding results indicate that pyrazole benzamides have a wide range of potential applications as medicinal drugs and diagnostic agents. As a small molecule inhibitor, it has a unique structure, low toxicity and excellent biological activity, so it is of great significance to develop antitumor drug preparations with new structure types.

Contents of the invention

In view of this, the object of the present invention is to provide pyrazole benzamide compounds and their preparation methods and applications, and the pyrazole benzamide compounds have better antitumor activity.

The pyrazole benzamide compound provided by the present invention has the structure of formula I:

The R ₁ and R ₂ are independently selected from hydrogen, alkyl, amino, nitro, halogen, alkoxy, trifluoromethyl, cyano, cycloalkyl, heterocyclyl, -COR ₃ , -COOR ₄ , -SO ₂ R ₅ , -NR ₆ R ₇ ;

Said R ₁ and R ₂ do not select hydrogen at the same time;

The R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are independently selected from hydrogen, alkyl, amino, halogen, alkoxy or trifluoromethyl.

In the present invention, the R ₁ is selected from -CF ₃ , -OH, or -O-CH ₃ ;

The R ₂ is selected from -NH ₂ , -OCH ₃ , -Cl, CH ₃ , -OH, or -NO ₂ .

In the present invention, the pyrazole benzamide compound has any one of formulas (I-1) to (I-12):

The present invention provides a kind of preparation method of pyrazole benzamide compounds described in the technical scheme, comprising the following steps:

reacting the compound of the formula II structure with the compound of the formula III structure to obtain the compound of the formula I structure;

R ₁ in formula II is the same as the substituent of R ₁ in the above technical scheme; R ₁ in formula III is the same as the substituent of R ₂ in the above technical scheme.

Wherein, the formula (II) 4-(3-oxo-3-phenylpropionyl)methyl benzoate is reacted with hydrazine monohydrochloride, and the obtained intermediate is then prepared by hydrolysis.

In the embodiment of the present invention, the pyrazole benzamide compound is preferably prepared according to the following reaction scheme:

The present invention provides an application of the pyrazole benzamide compound described in the above technical solution or the pyrazole benzamide compound prepared by the preparation method described in the above technical solution in the preparation of antitumor drugs.

The present invention provides an antineoplastic drug, comprising the pyrazole benzamide compound described in the above technical scheme or the pyrazole benzamide compound prepared by the preparation method described in the above technical scheme;

and medically acceptable excipients.

The above-mentioned antitumor drugs provided by the present invention can also be used in combination with other antitumor drugs.

The pyrazole benzamide compound provided by the present invention has the structure of formula I. Compared with the prior art, the pyrazole benzamide compound provided by the present invention has better anti-tumor activity, and can be used as a drug for the preparation of related tumor diseases.

Detailed ways

In order to further illustrate the present invention, the pyrazole benzamide compounds provided by the present invention and their preparation methods and applications are described in detail below in conjunction with the examples, but they should not be understood as limiting the protection scope of the present invention.

Example 1

Synthesis of N-(4-aminophenethyl)-4-(5-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-yl)benzamide:

t-BuOK (2.8 g, 25 mmol) was placed in a round bottom flask at 0°C, and THF (30 ml) was added and stirred for 5 min. In another flask, dimethyl terephthalate (2.91 g, 15 mmol) was dissolved in DME (100 ml), gradually added dropwise to trifluoromethyl acetophenone (1.52 ml, 10 mmol) and mixed. The mixture was gradually dropped into a round bottom flask at 0 °C, stirred at room temperature for 1 h, and the progress of the reaction was detected by TLC. A 4mol/L HCl solution was added dropwise to the mixture to adjust to pH=2. The aqueous layer was extracted with EtOAc, the organic layer was collected, dried over _Na2SO4 , and the solvent was removed in vacuo _. Methyl 4-(3-oxo-3-(3-(trifluoromethyl)phenyl)propionyl)benzoate was obtained by column chromatography (AcOEt/cyclohexane, 50:1 to 20:1) Solid (Compound 3). (yield 45%)

Compound 3 was dissolved in EtOH (30ml), added with hydrazine monohydrochloride (1.5eq), and refluxed at 80°C overnight. The mixture was extracted with EtOAc, the organic layer was collected, dried over Na2SO4, and the solvent was removed in vacuo. Compound 4 was obtained by column chromatography (EtOAc/cyclohexane, 10:1).

Compound 4 was dissolved in MeOH/H2O solution (2:1, 30ml), added KOH (5eq), and refluxed at 80°C for 2h for hydrolysis. EtOAc and aqueous extraction, the organic layer was extracted, added NaSO4 to dry, and the solvent was removed in vacuo to give compound 5.

Compound 5 was mixed with EDCI (1.5eq), HOBt (1.5eq), stirred at 0°C for 1h, added DIPEA (5eq) and 4-nitrophenethylamine (1eq), stirred r.t. overnight. The progress of the reaction was monitored by TLC. The mixture was extracted with EtOAc and aqueous solution, the organic layer was extracted, dried over Na2SO4, and the solvent was removed in vacuo. Compound 6 was obtained by column chromatography (EtOAc/cyclohexane, 1:1 to 1:2).

Dissolve compound 6 in MeOH/H ₂ O solution (2:1, 30ml), add excess Zn powder (5eq) and NH ₄ Cl (10eq), and stir at rt for 2h. Quenched by adding NaHCO ₃ . After filtering, washing with _EtOAc and aqueous solution, the organic layer was taken, dried over _Na2SO4 , and the solvent was removed in vacuo to give compound 7 as the final product.

Using trifluoromethyl acetophenone as raw material, the NMR data are as follows:

¹ H NMR (500MHz, DMSO-d ₆ ) δ8.54(s, 1H), 8.40(t, J=7.5Hz, 1H), 8.02(q, J=1.9Hz, 1H), 7.86–7.74(m, 5H), 7.72(dt, J=7.7, 1.9Hz, 1H), 7.03(s, 1H), 6.88–6.82(m, 2H), 6.50–6.44(m, 2H), 5.05(s, 2H), 3.47 (t,J=7.7Hz,2H),2.78–2.70(m,2H).

Example 2

Synthesis of N-(4-methoxyphenethyl)-4-(5-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-yl)benzamide:

Using trifluoromethyl acetophenone as raw material, refer to Example 1 for the synthesis method.

The NMR data are as follows:

1H NMR (400MHz, DMSO-d6) δ9.04(t, J＝6.0Hz, 1H), 8.19(d, J＝1.7Hz, 1H), 8.18–8.13(m, 1H), 8.01–7.92(m, 4H), 7.71–7.67(m, 2H), 7.49(s, 1H), 7.22(d, J=8.0Hz, 2H), 7.13(d, J=7.8Hz, 2H), 4.45(d, J=5.8 Hz,2H),2.27(s,3H).

Example 3

Synthesis of N-(4-chlorophenethyl)-4-(5-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-yl)benzamide:

The NMR data are as follows:

1H NMR (400MHz, DMSO-d6) δ8.60(t, J＝5.6Hz, 1H), 8.20(d, J＝1.4Hz, 1H), 8.16(ddd, J＝5.8, 3.4, 1.7Hz, 1H) ,7.96–7.88(m,4H),7.72–7.66(m,2H),7.47(s,1H),7.35(dt,J＝6.7,2.3Hz,2H),7.30–7.25(m,2H),3.50 (dd,J=7.5,5.7Hz,2H),2.86(t,J=7.3Hz,2H).

Example 4

Synthesis of N-(4-methylphenethyl)-4-(5-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-yl)benzamide:

1H NMR (500MHz, DMSO-d6) δ7.66(dd, J＝5.6, 3.4Hz, 2H), 7.55(t, J＝1.5Hz, 1H), 7.49(dd, J＝7.5, 2.0Hz, 1H) ,7.30–7.18(m,4H),7.09–7.03(m,1H),2.20(dd,J=10.0,1.0Hz,6H).

13C NMR (125MHz, Common NMR Solvents) δ157.70, 149.66, 137.72, 137.46, 137.39, 136.93, 134.43, 129.57, 126.97, 125.27, 123.62, 123.10, 123.04, 119.77, 118.40, 117.15, 114.42, 19.62, 18.75.

Example 5

Synthesis of N-(4-aminophenethyl)-4-(5-(4-hydroxyphenyl)-1H-pyrazol-3-yl)benzamide:

Using p-hydroxyacetophenone as raw material, see Example 1 for the synthesis method.

¹ H NMR(500MHz,DMSO-d6)δ9.63(s,1H),8.54(s,1H),7.80(s,4H),7.71–7.65(m,2H),6.96(s,1H),6.88 –6.80(m,4H),6.50–6.44(m,2H),5.05(s,2H),3.47(t,J＝5.2Hz,2H),2.77–2.71(m,2H).

Example 6

Synthesis of N-(4-aminophenethyl)-4-(5-(4-chlorophenyl)-1H-pyrazol-3-yl)benzamide:

Using p-chloroacetophenone as a raw material, see Example 1 for the synthesis method.

¹ H NMR(500MHz,DMSO-d6)δ8.54(s,1H),7.82(s,4H),7.78–7.71(m,2H),7.51–7.45(m,2H),7.01(s,1H) ,6.85(dt,J=7.4,1.1Hz,2H),6.50–6.44(m,2H),5.05(s,2H),3.47(t,J=7.7Hz,2H),2.74(tt,J=7.7 ,1.0Hz,2H).

Example 7

Synthesis of N-(4-aminophenethyl)-4-(5-(4-methoxyphenyl)-1H-pyrazol-3-yl)benzamide:

Taking p-methoxyacetophenone as raw material, see Example 1 for the synthetic method

¹ H NMR (500MHz, DMSO-d6) δ8.54(s, 1H), 7.82–7.73(m, 6H), 7.08–7.02(m, 2H), 6.97(s, 1H), 6.85(dt, J= 7.4,1.1Hz,2H),6.50–6.44(m,2H),5.05(s,2H),3.79(s,3H),3.47(t,J＝5.2Hz,2H),2.77–2.71(m,2H ).

Example 8

Synthesis of N-(4-aminophenethyl)-4-(5-(3-hydroxyphenyl)-1H-pyrazol-3-yl)benzamide:

With 3-hydroxyacetophenone as raw material, see Example 1 for the synthetic method

¹ H NMR(500MHz,DMSO-d6)δ9.12(s,1H),8.54(s,1H),7.81(s,4H),7.34–7.26(m,2H),7.16(q,J=1.4Hz ,1H),6.99(s,1H),6.88–6.78(m,3H),6.50–6.44(m,2H),5.05(s,2H),3.47(t,J＝5.2Hz,2H),2.74( tt,J=5.3,1.0Hz,2H).

Example 9

Synthesis of N-(4-hydroxyphenethyl)-4-(5-(4-hydroxyphenyl)-1H-pyrazol-3-yl)benzamide:

Taking p-hydroxyacetophenone as a raw material, the synthetic method is referring to Example 1

¹ H NMR (500MHz, DMSO-d6) δ9.63(s, 1H), 9.18(s, 1H), 8.54(s, 1H), 7.80(d, J=1.1Hz, 4H), 7.71–7.65(m ,2H),7.05–6.99(m,2H),6.94(s,1H),6.86–6.80(m,2H),6.71–6.65(m,2H),3.47(t,J＝5.2Hz,2H), 2.81(td,J=5.1,1.1Hz,2H).

Example 10

Synthesis of 4-(5-(4-hydroxyphenyl)-1H-pyrazol-3-yl)-N-(4-methoxyphenethyl)benzamide:

¹ H NMR (500MHz, DMSO-d6) δ9.63(s, 1H), 8.54(s, 1H), 7.81(s, 4H), 7.71–7.65(m, 2H), 7.13(dt, J=7.6, 1.1Hz, 2H), 6.94(s, 1H), 6.86–6.77(m, 4H), 3.79(s, 3H), 3.47(t, J=5.2Hz, 2H), 2.81(tt, J=5.2, 1.0 Hz,2H).

Example 11

Synthesis of 4-(5-(4-hydroxyphenyl)-1H-pyrazol-3-yl)-N-(4-nitrophenethyl)benzamide:

¹ H NMR (500MHz, DMSO-d6) δ9.63(s,1H),8.54(s,1H),8.17–8.11(m,2H),7.86–7.78(m,4H),7.71–7.65(m, 2H), 7.60–7.54(m, 2H), 6.96(s, 1H), 6.86–6.80(m, 2H), 3.47(t, J=7.7Hz, 2H), 2.86(dd, J=8.1, 7.0Hz ,2H).

Example 12

Synthesis of N-(4-chlorophenethyl)-4-(5-(4-hydroxyphenyl)-1H-pyrazol-3-yl)benzamide:

¹ H NMR (500MHz,DMSO-d6)δ9.63(s,1H),8.54(s,1H),7.86–7.78(m,4H),7.71–7.65(m,2H),7.36–7.30(m, 2H),7.29–7.23(m,2H),6.96(s,1H),6.86–6.80(m,2H),3.47(t,J=7.7Hz,2H),2.84(td,J=7.7,1.3Hz ,2H).

The selected tumor cells include colon cancer cell SW480, drug-resistant colon cancer cell SW620 and gastric cancer cell SGC7901. Cells in the logarithmic growth phase were taken, digested and counted, and the cell state was adjusted before plating. Seed in a 96-well plate at a density of 3000/well cells and place in an incubator overnight. (Note: The outer ring is filled with PBS.) After 18-24 hours, the drug administration operation is performed. Aspirate the cell medium, and add the culture solution containing a certain concentration of the drug. The experiment set up a blank group (solvent control group) and an experimental group. The drug concentrations in the experimental group were 0.05 μM, 0.1 μM, 1 μM, 5 μM, 10 μM, and 20 μM in sequence. Three auxiliary wells were set up for each group of experiments, and the positive control was pentafluorouracil, and the method was the same as that of the test compound.

After 72 hours of administration, 10 μL of MTT solution was added to each well, and placed in a cell culture incubator for further incubation for 2 hours. The absorbance value of each well at a wavelength of 492 nm was detected with a microplate reader.

The survival rate of cells in each well was calculated according to the absorbance value, and each experiment was repeated three times to calculate the IC ₅₀ value of the drug. The results are shown in Table 1.

Table 1 In vitro antitumor activity screening of test compounds

It can be seen from the above examples that the pyrazole benzamide derivatives prepared by the present invention have better antitumor activity.

The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims

Pyrazole benzamide compounds have the structure of formula I:

The R 1 and R 2 are independently selected from hydrogen, alkyl, amino, nitro, halogen, alkoxy, trifluoromethyl, cyano, cycloalkyl, heterocyclyl, -COR 3 , -COOR 4 , -SO 2 R 5 , -NR 6 R 7 ;

Said R 1 and R 2 do not select hydrogen at the same time;

The R 3 , R 4 , R 5 , R 6 and R 7 are independently selected from hydrogen, alkyl, amino, halogen, alkoxy or trifluoromethyl.
The pyrazole benzamide compound according to claim 1, wherein said R 1 is selected from -CF 3 , -OH, or -O-CH 3 ;

The R 2 is selected from -NH 2 , -OCH 3 , -Cl, CH 3 , -OH, or -NO 2 .
The pyrazole benzamide compound according to claim 1, wherein the pyrazole benzamide compound has any structure in formula (I-1)～(I-12):
A preparation method of pyrazole benzamide compounds described in any one of claims 1 to 3, comprising the following steps:

reacting the compound of the formula II structure with the compound of the formula III structure to obtain the compound of the formula I structure;
A use of the pyrazole benzamide compound according to any one of claims 1 to 3 or the pyrazole benzamide compound prepared by the preparation method according to claim 4 in the preparation of antitumor drugs.
An antitumor drug, comprising the pyrazole benzamide compound according to any one of claims 1 to 3 or the pyrazole benzamide compound prepared by the preparation method according to claim 4;

and medically acceptable excipients.