CN114262305A - Alkynyl-substituted thiazolinone-based-N-aryl benzamide compound and preparation method and application thereof - Google Patents

Abstract

The invention discloses an alkynyl-substituted thiazolone-N-arylbenzamide compound or a pharmaceutically acceptable salt and a preparation method thereof. The invention also discloses that the compound can be used to treat tumors such as bone and flesh It can significantly inhibit the activity of cancer cells and has broad application prospects in the field of medicine.

Description

Alkynyl-substituted thiazolone-N-arylbenzamide compounds and preparation method and application thereof

technical field

The invention belongs to the fields of synthetic medicine and biomedicine, and mainly relates to a preparation method and application of an alkynyl-substituted thiazolone-N-arylbenzamide compound or a pharmaceutically acceptable salt.

Background technique

Osteosarcoma, also known as osteosarcoma, is a malignant bone tumor that occurs more commonly in adolescents or children under the age of 20. [1] According to the epidemiological study of osteosarcoma, it is the most common bone tumor in children, about 5% of pediatric tumors. Osteosarcoma is still the highest mortality rate of malignant tumors in children and adolescents, second only to lymphoma and brain cancer, but early detection and timely treatment have greatly improved the survival rate of this disease.

The treatment methods for different types of osteosarcoma are usually combined after surgery. It is worth noting that the cancer metastasis rate of patients with osteosarcoma is as high as 80%, and 90% of the patients with metastases are lung metastasis. Among them, the recurrence rate is 20-30%, in situ or in situ tissue may recur, and the survival rate after recurrence, in the past few decades, despite the adjustment of increased doses, the number of changes and the combination of multiple chemotherapy drugs and other treatment methods , the survival rate did not improve.

At present, the research on the therapeutic targets of osteosarcoma mainly focuses on the family of cell surface receptor tyrosine kinases (PTKs) and intracellular signaling targets. For these targets, the main therapeutic drugs are divided into three categories: 1) immunomodulators; 2) receptor tyrosine kinase inhibitors; 3) intracellular signaling pathway inhibitors. In addition, new targets and treatment options for osteosarcoma include NK-kB ligands, folic acid inhibitors, etc., but they are all in the experimental research stage.

SUMMARY OF THE INVENTION

The inventors have noticed that in previous literature reports, compounds with thiazolinone skeleton structure have various pharmacological activities, such as the compound represented by formula (III) has anti-inflammatory activity and the compound represented by formula (IV) has Anti-AIDS Activity:

Before the present invention, there is no relevant report on the evaluation of osteosarcoma cell viability test for this type of scaffold. The inventors have found and proposed that the skeleton compound may generate a series of new compound molecules with meaningful structure and activity, which may provide a new source of compounds for biological activity screening.

The present invention provides an alkynyl-substituted thiazolone-N-arylbenzamide compound or a pharmaceutically acceptable salt, comprising 3-(3-(ethynylbenzene) represented by formula (I) )-4-thiazolinone group)-N-arylbenzamide compounds or pharmaceutically acceptable salts, 3-(3-aryl-4-thiazolinone group)- 5-ethynyl-N-arylbenzamide compounds or pharmaceutically acceptable salts.

Wherein, the 3-(3-(ethynylbenzene)-4-thiazolinone base)-N-arylbenzamide compound or pharmaceutically acceptable salt shown in formula (I), its structure is as follows:

in,

Ar is aryl, aromatic heterocycle, alkyl substituted aryl, halogen substituted aryl, alkyl substituted heteroaryl, halogen substituted heteroaryl, trifluoromethyl substituted aryl, nitro substituted Aryl, decalinyl, furanalkyl, alkenyl, alkynyl, diphenylalkyl, bis(4-bromophenyl)alkyl, trifluoromethyl substituted heteroaryl, nitro substituted heteroaryl ; Wherein, the aromatic heterocycle and heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole, pyrazine, etc.;

R ¹ is halogen, alkoxy, alkyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen, etc.;

R ² is alkyl, hydrogen, etc.;

R ³ is halogen, alkoxy, alkyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen, etc.;

n=0;1;2;3;4;5;6.

Preferably,

Ar is aryl, aromatic heterocycle, C1-C10 alkyl substituted aryl, halogen substituted aryl, C1-C10 alkyl substituted heteroaryl, halogen substituted heteroaryl, trifluoromethyl substituted aryl aryl, nitro-substituted aryl, naphthylmethyl, furylmethyl, allyl, propargyl, benzhydryl, bis(4-bromophenyl)methyl, trifluoromethyl substituted heteroaryl , nitro-substituted heteroaryl; wherein, the aromatic heterocycle and heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole, Pyrazine;

R ¹ is halogen, C1-C10 alkoxy, methyl, ethyl, isopropyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

R ² is methyl or hydrogen;

R ³ is halogen, C1-C10 alkoxy, methyl, ethyl, isopropyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen.

n=1;2;3;4;5;6.

Wherein, the 3-(3-aryl-4-thiazolinone)-5-ethynyl-N-arylbenzamide compound or pharmaceutically acceptable salt represented by formula (II) has the following structure :

in,

Ar is aryl, aromatic heterocycle, alkyl substituted aryl, halogen substituted aryl, alkyl substituted heteroaryl, halogen substituted heteroaryl, trifluoromethyl substituted aryl, nitro substituted Aryl, decalinyl, furanalkyl, alkenyl, alkynyl, diphenylalkyl, bis(4-bromophenyl)alkyl, trifluoromethyl substituted heteroaryl, nitro substituted heteroaryl ; Wherein, the aromatic heterocycle and heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole, pyrazine, etc.;

R ¹ is halogen, alkoxy, alkyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen, etc.;

R ² is alkyl, hydrogen, etc.;

n=0;1;2;3;4;5;6.

Preferably,

Ar is aryl, aromatic heterocycle, C1-C10 alkyl substituted aryl, halogen substituted aryl, C1-C10 alkyl substituted heteroaryl, halogen substituted heteroaryl, trifluoromethyl substituted aryl aryl, nitro-substituted aryl, naphthylmethyl, furylmethyl, allyl, propargyl, benzhydryl, bis(4-bromophenyl)methyl, trifluoromethyl substituted heteroaryl , nitro-substituted heteroaryl; wherein, the aromatic heterocycle and heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole, Pyrazine;

R ¹ is halogen, C1-C10 alkoxy, methyl, ethyl, isopropyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

R ² is methyl or hydrogen.

n=1;2;3;4;5;6.

The 3-(3-(ethynylbenzene)-4-thiazolinone group)-N-arylbenzamide compound or 3-(3-aryl-4-thiazolinone group)- 5-ethynyl-N-arylbenzamide compounds or pharmaceutically acceptable salts, including but not limited to the following:

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-N-pyrazobutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-N-phenylpropyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-N-phenylethyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-N-indolebutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-N-pyrazolylbutyl)benzamide;

3-(3-(2-Methyl-5-ethynylbenzene)-4(5-methyl)-thiazolinone)-N-(4-N-indolebutyl)benzamide;

3-(3-(2-methoxybenzene)-4-thiazolinone)-5-ethynyl-N-(4-phenylbutyl)benzamide;

3-(3-(2-Ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Ethynylbenzene)-4-thiazolinone)-5-fluoro-N-(4-phenylbutyl)benzamide;

3-(3-(2-Ethynylbenzene)-4-thiazolinone)-5-methyl-N-(4-phenylbutyl)benzamide;

3-(3-(2-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-(2-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-fluoro)phenylbutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-5-fluoro-N-(4-phenylbutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-5-methyl-N-(4-phenylbutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-(3-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-fluoro)phenylbutyl)benzamide;

3-(3-(4-Ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(4-Ethynylbenzene)-4-thiazolinone)-5-fluoro-N-(4-phenylbutyl)benzamide;

3-(3-(4-Ethynylbenzene)-4-thiazolinone)-5-methyl-N-(4-phenylbutyl)benzamide;

3-(3-(4-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-(4-Ethynylbenzene)-4-thiazolinone)-N-(4-(4-fluoro)phenylbutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-benzopyrrolylbutyl)benzamide;

3-(3-(2-Methyl-4-ethynylbenzene)-4-thiazolinone)-N-(4-carbazolylbutyl)benzamide;

3-(3-(2-Methyl-5-ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-5-fluoro-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-5-methyl-N-(4-phenylbutyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-(2-Methoxy-5-ethynylbenzene)-4-thiazolinone)-N-(4-(4-fluoro)phenylbutyl)benzamide;

3-(3-Phenyl-4-thiazolinone)-5-ethynyl-N-(4-phenylbutyl)benzamide;

3-(3-Phenyl-4-thiazolinone)-5-ethynyl-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-phenyl-4-thiazolinone)-5-ethynyl-N-(4-(4-fluoro)phenylbutyl)benzamide;

3-(3-(2-methoxybenzene)-4-thiazolinone)-5-ethynyl-N-(4-(4-chloro)phenylbutyl)benzamide;

3-(3-(2-Methoxybenzene)-4-thiazolinonyl)-5-ethynyl-N-(4-(4-fluoro)phenylbutyl)benzamide.

In the present invention, the acid addition salt formed by the compound and acid; wherein, the acid is hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, salicylic acid, tartaric acid, lactic acid, citric acid, methanesulfonic acid, p- Toluenesulfonic acid, maleic acid, pyruvic acid or succinic acid, etc.

The present invention also provides a pharmaceutical composition comprising the alkynyl-substituted thiazolone-N-arylbenzamide compound or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

Wherein, the pharmaceutical composition is formulated into pills, capsules, creams, gels, excipients, injectable fluids, aerosols, syrups or transdermal patches.

The present invention also provides the use of the alkynyl-substituted thiazolone-N-arylbenzamide compounds or pharmaceutically acceptable salts, or pharmaceutical compositions in the preparation of medicaments for preventing and/or treating tumors application.

Wherein, the alkynyl-substituted thiazolone-N-arylbenzamide compounds or pharmaceutically acceptable salts, or pharmaceutical compositions can be used to inhibit tumor cell proliferation, growth, migration, infiltration, metastasis and recurrence, or promote tumor cell apoptosis.

Wherein, the tumor includes osteosarcoma, leukemia, pancreatic cancer, lung cancer and the like.

The osteosarcoma cells include SJSA-1 type osteosarcoma cells, MNNG/HOS type osteosarcoma cells, U-2-OS type osteosarcoma cells, MG63 type osteosarcoma cells, HOS type osteosarcoma cells, and 143B type osteosarcoma cells.

The lung cancer cells include A549 type lung cancer cells.

The leukemia includes Jurkat leukemia cells.

The pancreatic cancer cells include PANC-1 pancreatic cancer cells.

The present invention provides a 3-(3-(ethynylbenzene)-4-thiazolinone)-N-arylbenzamide compound represented by formula (I) or a pharmaceutically acceptable salt. The synthetic method specifically comprises the steps as follows, comprising the following four reaction formulas (A), (B), (C), (D):

In the first step, Sonogashira coupling reaction is carried out with p-iodoaniline and trimethylsilyl acetylene to obtain p-trimethylsilyl ethynyl aniline, and then the trimethylsilyl group is removed with potassium carbonate to obtain the product p-ethynyl aniline .

In the second step, p-ethynylaniline (10 mmol, 1.0 eq) was reacted with methyl 3-formaldehyde benzoate (10 mmol, 1.0 eq) under toluene reflux for 30 minutes, and the corresponding imine compound was generated at this time. Subsequently, it was cooled to room temperature, and thioethanol (10 mmol, 1.0 eq) was added to the reaction system to continue refluxing, and the reaction was cooled to room temperature after 5 h. :1～1:1) separation and purification to obtain ester compounds.

In the third step, the ester compound (5mmol, 1.0eq) obtained in the second step was reacted with LiOH (5.25mmol, 1.05eq) in a solvent of methanol:water (4:1) at room temperature for 20 hours. Monitor the complete consumption of the substrate. After the reaction, remove the methanol in the solvent by rotary evaporation, then pour the remaining reaction solution into 50 mL of water, add 3N HCl dropwise to acidify, and a large amount of white solids are precipitated. Use pH test paper to monitor until pH=1 . The white solid was filtered off with a Buchner funnel, and the white solid was dissolved in ethyl acetate, dried over anhydrous sodium sulfate, and rotary-evaporated to obtain a carboxylic acid product.

In the fourth step, the carboxylic acid product (1.5 mmol, 1.0 eq) obtained in the third step and 1-hydroxybenzotriazole (ie, HOBt, 3.0 mmol, 2.0 eq) were dissolved in 20 mL of tetrahydrofuran, and under ice bath conditions, added N,N-diisopropylethylamine (DIPEA, 4.5 mmol, 3.0 eq), after stirring for 20 minutes, the compound amine was added, and stirring was continued for 10 minutes. Subsequently, the condensing agent 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI, 3 mmol, 2.0 eq) was added in batches, and after the reaction for two hours, the spot plate was monitored, and the raw materials were completely consumed. . An appropriate amount of saturated sodium bicarbonate was sequentially added to the reaction solution, washed with saturated brine, and extracted three times with ethyl acetate. The crude product obtained by rotary evaporation was subjected to column chromatography (petroleum ether:ethyl acetate=3:1-1: 1) The target product is finally obtained.

Wherein, in the reaction formulas (A), (B), (C) and the reaction formula (D), the definitions of Ar, R ¹ , R ² and R ³ are the same as those of formula (I).

The present invention also provides a 3-(3-aryl-4-thiazolinone)-5-ethynyl-N-arylbenzamide compound represented by formula (II) or a pharmaceutically acceptable compound The synthetic method of the salt, specifically comprises steps as follows, comprises following four reaction formulas (E), (F), (G), (H):

In the first step, a methyl ester group in methyl 5-iodoisophthalate is reduced to an alcohol with sodium borohydride, and then the alcohol is oxidized to an aldehyde under manganese dioxide conditions. The obtained compound is subjected to Sonogashira coupling reaction with trimethylsilyl acetylene to generate methyl 3-carbaldehyde-5-trimethylsilyl ethynyl benzoate, followed by removal of the trimethylsilyl group with potassium carbonate to obtain product 3 -Methyl formaldehyde-5-ethynylbenzoate.

In the second step, substituted aniline (10mmol, 1.0eq) was reacted with methyl 3-carbaldehyde-5-ethynylbenzoate (10mmol, 1.0eq) under toluene reflux for 30 minutes, and the corresponding imine compound was generated at this time. . Subsequently, it was cooled to room temperature, and thioethanol (10 mmol, 1.0 eq) was added to the reaction system to continue refluxing, and the reaction was cooled to room temperature after 5 h. :1～1:1) separation and purification to obtain ester compounds.

In the third step, the ester compound (5mmol, 1.0eq) obtained in the second step was reacted with LiOH (5.25mmol, 1.05eq) in a solvent of methanol:water (4:1) at room temperature for 20 hours. Monitor the complete consumption of the substrate. After the reaction, remove the methanol in the solvent by rotary evaporation, then pour the remaining reaction solution into 50 mL of water, add 3N HCl dropwise to acidify, and a large amount of white solids are precipitated. Use pH test paper to monitor until pH=1 . The white solid was filtered off with a Buchner funnel, and the white solid was dissolved in ethyl acetate, dried over anhydrous sodium sulfate, and rotary-evaporated to obtain a carboxylic acid product.

In the fourth step, the carboxylic acid product (1.5 mmol, 1.0 eq) obtained in the third step and 1-hydroxybenzotriazole (ie, HOBt, 3.0 mmol, 2.0 eq) were dissolved in 20 mL of tetrahydrofuran, and under ice bath conditions, added N,N-diisopropylethylamine (DIPEA, 4.5 mmol, 3.0 eq), after stirring for 20 minutes, the compound amine was added, and stirring was continued for 10 minutes. Subsequently, the condensing agent 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI, 3 mmol, 2.0 eq) was added in batches, and after the reaction for two hours, the spot plate was monitored, and the raw materials were completely consumed. . An appropriate amount of saturated sodium bicarbonate was sequentially added to the reaction solution, washed with saturated brine, and extracted three times with ethyl acetate. The crude product obtained by rotary evaporation was subjected to column chromatography (petroleum ether:ethyl acetate=3:1～1: 1) The target product is finally obtained.

Wherein, in the reaction formulas (E), (F), (G) and the reaction formula (H), the definitions of Ar, R ¹ and R ² are the same as those of the formula (II).

The beneficial effects of the invention are as follows: the invention discloses an alkynyl-substituted thiazolone-N-arylbenzamide compound or a pharmaceutically acceptable salt, which has inhibitory activity on tumor cells and can be used for The treatment of anti-osteosarcoma and other related diseases is of great significance in the research of tumor diseases.

Description of drawings

FIG. 1 is a schematic diagram of ¹ H NMR ( FIG. 1A ) and ¹³ C NMR ( FIG. 1B ) of the product A-1 obtained in Example 1. FIG.

FIG. 2 is a schematic diagram of ¹ H NMR ( FIG. 2A ) and ¹³ C NMR ( FIG. 2B ) of the product A-2 obtained in Example 2. FIG.

3 is a schematic diagram of ¹ H NMR ( FIG. 3A ) and ¹³ C NMR ( FIG. 3B ) of the product A-3 obtained in Example 3. FIG.

FIG. 4 is a schematic diagram of ¹ H NMR ( FIG. 4A ) and ¹³ C NMR ( FIG. 4B ) of the product A-4 obtained in Example 4. FIG.

FIG. 5 is a schematic diagram of ¹ H NMR ( FIG. 5A ) and ¹³ C NMR ( FIG. 5B ) of the product A-5 obtained in Example 5. FIG.

FIG. 6 is a schematic diagram of ¹ H NMR ( FIG. 6A ) and ¹³ C NMR ( FIG. 6B ) of the product A-6 obtained in Example 6. FIG.

FIG. 7 is a schematic diagram of ¹ H NMR ( FIG. 7A ) and ¹³ C NMR ( FIG. 7B ) of the product A-7 obtained in Example 7. FIG.

FIG. 8 is a schematic diagram of ¹ H NMR ( FIG. 8A ) and ¹³ C NMR ( FIG. 8B ) of the product A-8 obtained in Example 8. FIG.

FIG. 9 is a schematic diagram of ¹ H NMR ( FIG. 9A ) and ¹³ C NMR ( FIG. 9B ) of the product A-9 obtained in Example 9. FIG.

FIG. 10 is a schematic diagram of ¹ H NMR ( FIG. 10A ) and ¹³ C NMR ( FIG. 10B ) of the product A-10 obtained in Example 10.

Detailed ways

The present invention will be further described in detail with reference to the following specific embodiments and accompanying drawings, and the protection content of the present invention is not limited to the following embodiments. Variations and advantages that can occur to those skilled in the art without departing from the spirit and scope of the inventive concept are included in the present invention, and the appended claims are the scope of protection.

For the preparation methods of compounds 1-10 in the following examples of the present invention, reference is made to the preparation of compounds represented by formula (I) or formula (II) mentioned above. Determination methods: low-resolution mass spectrometry (molecular weight) and nuclear magnetic instruments (hydrogen spectrum and carbon spectrum).

Example 1:

The ¹ H NMR and ¹³ C NMR spectra of nuclear magnetic resonance are shown in Figure 1:

¹ H NMR (400MHz, Chloroform-d) δ7.78(d, J=6.4Hz, 1H), 7.57(d, J=7.8Hz, 1H), 7.42(t, J=6.2Hz, 1H), 7.29– 7.05(m,9H), 6.71–6.60(m,1H), 6.08(s,1H), 3.92–3.75(m,3H), 3.73–3.59(m,3H), 3.40–3.20(m,2H), 2.91(s, 1H), 2.57(q, J=7.0Hz, 2H), 1.56(dt, J=24.2Hz, 7.6 Hz, 4H). ¹³ C NMR (100MHz, CDCl ₃ )δ171.43, 166.73, 155.35, 142.12 ,139.21,134.96,133.76, 133.67,130.47,128.79,128.44,128.36,127.49,126.77,125.84,125.29,114.66,112.16,82.41, 77.60,77.28,77.03,76.96,64.30,55.92,39.97,35.49,33.13,29.11 , 28.71.

Example 2:

Nuclear magnetic resonance ¹ H NMR and ¹³ C NMR spectra are shown in Figure 2:

¹ H NMR(400MHz, Chloroform-d)δ7.65(s,1H),7.52(s,1H),7.39(d,J=7.5Hz,1H),7.31-7.16(m,7H),7.11(t , J＝7.2Hz, 5H), 3.95(d, J＝15.7Hz, 1H), 3.82(d, J＝15.9Hz, 1H), 3.36 (d, J＝6.1Hz, 2H), 2.96(s, 1H) ), 2.59(t, J=6.9Hz, 3H), 1.68–1.51(m, 8H), 1.26(s, 2H).

¹³ C NMR (101MHz, CDCl ₃ ) δ 166.55, 142.00, 135.07, 130.49, 128.98, 128.39, 127.42, 125.89, 82.64, 78.13, 40.02, 35.47, 31.44, 30.20, 29.71, 29.06, 28

Example 3:

Nuclear magnetic resonance ¹ H NMR and ¹³ C NMR spectra are shown in Figure 3:

¹ H NMR (400MHz, Chloroform-d) δ7.65(s, 1H), 7.52(d, 1H), 7.39(d, J=7.5Hz, 1H), 7.32 -7.17(m, 7H), 7.10(q , J=12.3Hz, 9.7Hz, 6H), 5.99(s, 1H), 3.95(d, J=15.7Hz, 1H), 3.82(d, J=15.9Hz, 1H), 3.36(d, J=6.1 Hz, 2H), 2.96(s, 1H), 2.59(t, J=6.9Hz, 3H), 1.68–1.49(m, 8H), 1.26(s, 2H), 0.81(s, 1H).

¹³ C NMR (101MHz, CDCl ₃ )δ169.35, 135.89, 134.45, 130.13, 128.59, 128.27, 127.79, 121.46, 121.01, 119.31, 109.37, 101.15, 45.82, 39.42, 27.46, 26.8

Example 4:

Nuclear magnetic resonance ¹ H NMR and ¹³ C NMR spectra are shown in Figure 4:

¹ H NMR (400MHz, Chloroform-d) δ 7.63 (s, 1H), 7.55 (d, J=7.8 Hz, 1H), 7.43 (d, J=7.5 Hz, 1H), 7.38 (d, J=7.5 Hz, 1H), 7.22 (dt, J=15.5Hz, 8.1Hz, 3H), 7.11 (t, J=7.4Hz, 2H), 7.06– 6.97 (m, 3H), 6.77 (t, J=8.3Hz, 2H), 6.70–6.50(m, 3H), 4.07(t, J=6.4Hz, 3H), 3.90–3.72(m, 3H), 3.67(s, 3H), 3.27(q, J=6.1Hz, 3H) ), 1.83–1.75 (m, 3H), 1.56–1.34 (m, 3H).

¹³ C NMR(101MHz,CDCl3)δ170.37,165.69,156.32,153.80,150.25,138.39,134.93, 133.80,129.48,127.86,127.47,126.69,126.30,125.61,124.88,124.78,120.46,120.01,118.30, 116.28,116.04, 115.03, 114.80, 111.76, 111.68, 108.32, 100.14, 63.10, 55.23, 44.81, 38.43, 32.07, 26.43, 26.04.

Example 5:

Nuclear magnetic resonance ¹ H NMR and ¹³ C NMR spectra are shown in Figure 5:

¹ H NMR (400MHz, Chloroform-d) δ7.64 (d, J=12.8Hz, 1H), 7.53 (d, J=7.4Hz, 0H), 7.36 (d, J=7.9Hz, 1H), 7.18 ( d, J＝7.6Hz, 3H), 7.08(d, J＝8.0Hz, 4H), 5.99(s, 0H), 4.18(q, J＝7.4 Hz, 1H), 4.02(q, J＝7.2Hz, 1H), 3.29(d, J＝7.0Hz, 2H), 2.55(t, J＝7.6Hz, 2H), 2.19–2.07(m, 1H), 1.93(d, J＝10.2Hz, 1H), 1.67( d,J=6.9Hz,1H),0.79(q,J=10.0Hz,8.3Hz,1H).

¹³ C NMR (101MHz, CDCl ₃ )δ166.65, 142.05, 131.49, 131.46, 128.43, 128.37, 125.87, 120.72, 39.99, 35.49, 29.71, 29.14, 28.69, 20.16, 19.90.

Example 6:

Nuclear magnetic resonance ¹ H NMR and ¹³ C NMR spectra are shown in Figure 6:

¹ H NMR (400MHz, Chloroform-d) δ7.66(d, J=9.4Hz, 2H), 7.62(s, 1H), 7.32-7.28(m, 2H), 7.22-7.16(m, 4H), 6.93 (d, J=7.7Hz, 1H), 6.88–6.79 (m, 2H), 6.09 (s, 1H), 5.95 (d, J=12.7Hz, 1H), 3.97 (d, J=15.5Hz, 1H) ,3.90(d,J＝15.7Hz,1H),3.82(s,3H),3.43(q,J＝6.8Hz,2H),3.12(s,1H),2.66(t,J＝7.4Hz,2H) ,1.71–1.61(m,4H).

¹³ C NMR(101MHz,CDCl3)δ171.32,165.84,154.80,141.95,140.50,135.25,133.89, 130.60,129.89,128.39,126.94,125.91,125.03,122.87,121.04,112.22,82.15,78.73,63.97,55.79, 40.03, 35.44, 33.08, 29.10, 28.66.

Example 7:

Nuclear magnetic resonance ¹ H NMR. The ¹³ C NMR spectrum is shown in Figure 7:

¹ H NMR (400MHz, Chloroform-d) δ7.70(s, 1H), 7.58(d, J=7.6Hz, 1H), 7.43(d, J=7.8 Hz, 1H), 7.36(t, 2H), 7.32–7.26 (m, 3H), 7.24–7.12 (m, 5H), 6.14 (s, 1H), 6.02 (t, J=5.7Hz, 1H), 3.98 (d, J=15.9Hz, 1H), 3.88 (d, J=15.8Hz, 1H), 3.45 (q, J=6.7Hz, 2H), 3.04 (s, 1H), 2.67 (t, J=7.3Hz, 2H), 1.75–1.62 (m, 4H) .

¹³ C NMR(101MHz,CDCl ₃ )δ171.32,165.84,154.80,141.95,140.50,135.25,133.89, 130.60,129.89,128.39,126.94,125.91,125.03,122.87,121.04,112.22,82.15,78.73,63.97,55.79, 40.03 ,35.44,33.08,29.10,28.66.

Example 8:

Nuclear magnetic resonance ¹ H NMR. The ¹³ C NMR spectrum is shown in Figure 8:

¹ H NMR(400MHz, Chloroform-d)δ7.74(s,1H),7.61(d,J=7.1Hz,1H),7.42(s,1H),7.32(s,2H),7.26(s,2H) ), 7.21–7.15(m, 2H), 7.14–7.01(m, 3H), 6.95(s, 1H), 6.17(s, 1H), 6.06(s, 1H), 4.13–4.05(m, 3H), 3.88(d,J=15.8Hz,1H),3.76(d,J=15.8Hz,1H),3.37–3.20(m,3H), 2.99(s,1H),1.90–1.67(m,3H),1.56 –1.42(m,3H).

¹³ C NMR(101MHz,CDCl3)δ171.00,166.65,139.71,139.14,137.34,135.25,130.82, 129.63,129.32,129.21,128.95,127.53,126.08,126.00,123.20,105.57,82.49,78.40,64.95,51.26, 39.65, 33.43, 28.06, 25.99.

Example 9:

Nuclear magnetic resonance ¹ H NMR. The ¹³ C NMR spectrum is shown in Figure 9:

¹ H NMR (400MHz, Chloroform-d) δ 7.66 (t, J=1.5Hz, 1H), 7.53 (dt, J=7.6Hz, 1.3Hz, 1H), 7.40 (dt, J=7.6Hz, 1.3Hz ,1H),7.36–7.29(m,3H),7.29–7.24(m,2H),7.25–7.16(m,3H),7.12(ddd,J=8.0Hz,2.0Hz,1.3Hz,1H),6.22 (s,1H),6.11(s,1H),3.96(dd,J＝15.9Hz,1.5Hz,1H),3.85(d,J＝15.9Hz,1H),3.75–3.64(m,2H),3.06 (s,1H),2.91(t,J＝6.9Hz,1H).

¹³ C NMR(101MHz,CDCl3)δ170.94,166.63,139.89,138.81,137.35,135.42,130.85,129.73, 129.22,128.75,127.21,126.68,126.06,125.82,123.27,82.48,78.35,64.92,58.41,41.20,35.59, 33.37, 18.41.

Example 10:

Nuclear magnetic resonance ¹ H NMR. The ¹³ C NMR spectrum is shown in Figure 10:

¹ H NMR(400MHz, Chloroform-d)δ7.65(s,1H),7.49(d,J=7.7Hz,1H),7.39(d,J=7.8Hz,1H),7.35-7.22(m,5H) ), 7.23–7.14(m, 4H), 7.12(d, J=8.1Hz, 1H), 6.27(s, 1H), 6.10(s, 1H), 3.96(s, 0H), 3.81(s, 0H) ,3.43(q,J＝6.6Hz,2H),3.03(s,1H),2.68(t,J＝7.5Hz,2H),1.92(p,J＝7.2Hz,3H).

¹³ C NMR(101MHz,CDCl ₃ )δ170.99,166.54,141.48,139.81,137.38,135.37,130.84, 129.57,129.22,128.93,128.57,128.41,127.25,126.09,125.87,123.26,82.49,78.43,64.92,39.97, 33.54 ,33.39,30.97.

Example 11: Anti-tumor cell activity test experiment:

1. Seeding cells:

1) Prepare a single cell suspension with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and inoculate 2500 SJSA-1 osteosarcoma cells per well into a 96-well cell culture plate. Volume 100 μl.

2) A single cell suspension was prepared with DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and 2500 MNNG/HOS osteosarcoma cells per well were inoculated into a 96-well cell culture plate. Volume 100 μl.

3) A single cell suspension was prepared with McCoy'5A medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and 2500 U-2-OS osteosarcoma cells per well were inoculated into 96-well cell culture plate, with a volume of 100 μl per well.

4) Prepare a single cell suspension with DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and inoculate 2500 MG63 osteosarcoma cells per well into a 96-well cell culture plate, with a volume of 100 μl per well .

5) Prepare a single cell suspension with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and inoculate 2500 HOS osteosarcoma cells per well into a 96-well cell culture plate, with a volume of 100 μl per well .

6) Prepare a single cell suspension with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and inoculate 2500 143B osteosarcoma cells per well into a 96-well cell culture plate, with a volume of 100 μl per well .

7) A single cell suspension was prepared with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and 2500 A549 lung cancer cells per well were inoculated into a 96-well cell culture plate, with a volume of 100 μl per well.

8) A single cell suspension was prepared with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and 2500 Jurkat leukemia cells per well were inoculated into a 96-well cell culture plate, with a volume of 100 μl per well.

9) A single cell suspension was prepared with 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and 2500 PANC-1 pancreatic cancer cells per well were inoculated into a 96-well cell culture plate, with the volume of each well. 100 μl.

2. Administration: The compounds of formula (I) and (II) synthesized in the foregoing embodiments of the present invention are respectively prepared into a mother liquor with a final concentration of 25 mM with DMSO, diluted to 1 uM with complete medium, and acted on cells, and three are set. Duplicate holes.

3. Cultivation: 5% CO ₂ , 37°C saturated humidity incubator for 72 hours.

4. Coloring: After culturing for 72 hours, aspirate the medium, add 100 μl of complete medium and 10 μl of CCK8 to each well, and incubate at 37°C for 4 hours.

5. Colorimetry: select the wavelengths of 620 and 450 nm, measure the optical density (OD) value of each well on a microplate reader, and record the results.

6. The absorbance value at 450 nm - the absorbance value at 620 nm (background absorbance value) in the same well is used as the final absorbance, and is substituted into the following formula.

7. Cell proliferation activity (%)=[A (dosing)-A (blank)]/[A (0 dosing)-A (blank)]×100.

A (dosed): with osteosarcoma cells, CCK solution (CellCountingKit-8, is a WST-8 based (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4- Nitrophenyl)-5-(2,4-disulfobenzene)-2H-tetrazolium monosodium salt) rapid and high-sensitivity detection reagent for cell proliferation and cytotoxicity) and drug solution (drug solution refers to the compound to be tested) The absorbance of the wells of the sample DMSO solution)

A (blank): absorbance of wells with medium and CCK solution without cells

A (0 dosing): absorbance of wells with cells, CCK solution but no drug solution

8. The experimental results show that: under the action of the compounds of formula (I) and (II) of the present invention at a concentration of 1 μM, the survival rates of 6 different subtypes of osteosarcoma cells are shown in Table 1 below. It shows that among the 10 compounds, 3 compounds A-2, A-6, A-7 showed better inhibitory activity on osteosarcoma cells 143B, SJSA-1, and HOS; Osteosarcoma cells MNNG/HOS and MG63 showed very good inhibitory activity.

Table 1

9. The experimental results show that: under the action of the compounds of formula (I) and (II) of the present invention at a concentration of 1 μM, the survival rate of lung cancer cell A549 is shown in Table 2 below. This shows that among the 10 compounds, 3 compounds A-2, A-6 and A-7 showed good inhibitory activity on lung cancer cell A549.

Table 2

10. The experimental results show that: under the action of the compounds of formula (I) and (II) of the present invention at a concentration of 1 μM, the survival rate of the leukemia cell line Jurkat is shown in Table 3 below. This shows that, among the 10 compounds, 5 compounds A-1, A-2, A-6, A-7, A-8 showed better inhibitory activity on the leukemia cell Jurkat.

table 3

11. The experimental results show that: under the action of the compounds of formula (I) and (II) of the present invention at a concentration of 1 μM, the survival rate of pancreatic cancer cell line PANC-1 is shown in Table 4 below. This shows that among these 10 compounds, 2 compounds A-6 and A-7 showed good inhibitory activity on pancreatic cancer cell line PANC-1.

Table 4

The protection content of the present invention is not limited to the above embodiments. Variations and advantages that can occur to those skilled in the art without departing from the spirit and scope of the inventive concept are included in the present invention, and the appended claims are the scope of protection.

Claims (10)

1. A thiazolinone-N-aryl benzamide compound or pharmaceutically acceptable salt substituted with alkynyl is characterized in that the structure is respectively shown as formula (I) and formula (II):

wherein,

ar is aryl, aromatic heterocycle, alkyl substituted aryl, halogen substituted aryl, alkyl substituted heteroaryl, halogen substituted heteroaryl, trifluoromethyl substituted aryl, nitro substituted aryl, naphthylalkyl, furyl, alkenyl, alkynyl, diphenylalkyl, di (4-bromophenyl) alkyl, trifluoromethyl substituted heteroaryl, nitro substituted heteroaryl; wherein, the aromatic heterocyclic ring and the heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole and pyrazine;

R¹halogen, alkoxy, alkyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

R²is alkyl or hydrogen;

R³halogen, alkoxy, alkyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

n＝0；1；2；3；4；5；6。

2. substituted alkynyl thiazolinone-N-arylbenzamides or pharmaceutically acceptable salts according to claim 1 wherein Ar is aryl, heteroaromatic, C1-C10 alkyl substituted aryl, halo substituted aryl, C1-C10 alkyl substituted heteroaryl, halo substituted heteroaryl, trifluoromethyl substituted aryl, nitro substituted aryl, naphthylmethyl, furylmethyl, allyl, propargyl, benzhydryl, bis (4-bromophenyl) methyl, trifluoromethyl substituted heteroaryl, nitro substituted heteroaryl; wherein, the aromatic heterocyclic ring and the heteroaryl are respectively selected from indole, benzotriazole, thiophene, furan, pyrrole, bislactam, piperazine, imidazole, oxazole and pyrazine;

R¹is halogen, C1-C10 alkoxy, methyl, ethyl, isopropyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

R²is methyl or hydrogen;

R³halogen, alkoxy, methyl, ethyl, isopropyl, nitro, cyano, trifluoromethyl, phenyl, hydrogen;

n＝1；2；3；4；5；6。

3. the substituted alkynyl thiazolinone-N-aryl benzamide compound or pharmaceutically acceptable salt according to claim 1 or 2 selected from the group consisting of:

3- (3- (3-ethynylbenzene) -4-thiazolinonyl) -N- (4-phenylbutyl) benzamide;

3- (3- (2-methoxy-5-ethynylbenzene) -4-thiazolinonyl) -N- (4-phenylbutyl) benzamide;

3- (3- (2-methyl-4-ethynylbenzene) -4-thiazolinonyl) -N- (4-phenylbutyl) benzamide;

3- (3- (2-methyl-4-ethynylbenzene) -4-thiazolinonyl) -N- (4-N-indolylbutyl) benzamide;

3- (3- (2-methyl-4-ethynylbenzene) -4-thiazolinonyl) -N- (4-N-pyrazolylbutyl) benzamide;

3- (3- (3-ethynylbenzene) -4-thiazolinonyl) -N- (4-N-pyrazolylbutyl) benzamide;

3- (3- (3-ethynylbenzene) -4-thiazolinonyl) -N- (4-N-phenylpropyl) benzamide;

3- (3- (3-ethynylbenzene) -4-thiazolinonyl) -N- (4-N-phenylethyl) benzamide;

3- (3- (2-methyl-5-ethynylbenzene) -4 (5-methyl) -thiazolinonyl) -N- (4-N-indolylbutyl) benzamide;

3- (3- (2-methoxybenzene) -4-thiazolinonyl) -5-ethynyl-N- (4-phenylbutyl) benzamide.

4. The substituted alkynyl thiazolinone-N-aryl benzamide compound or the pharmaceutically acceptable salt according to claim 1 or 2 characterized in that the compound forms an acid addition salt with an acid; wherein the acid is hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, salicylic acid, tartaric acid, lactic acid, citric acid, methanesulfonic acid, p-toluenesulfonic acid, maleic acid, pyruvic acid, or succinic acid.

5. A pharmaceutical composition comprising the substituted alkynyl thiazolinone-N-aryl benzamide compound or a pharmaceutically acceptable salt according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is formulated as a pill, capsule, cream, gel, excipient, injectable fluid, aerosol, syrup, or transdermal patch.

7. Use of a substituted alkynyl thiazolinone-N-aryl benzamide compound or a pharmaceutically acceptable salt according to any one of claims 1 to 3 or a pharmaceutical composition according to claim 5 or 6 in the preparation of a medicament for the prophylaxis and/or treatment of tumors.

8. The use according to claim 7, wherein the substituted alkynyl thiazolinone-N-aryl benzamide compound or pharmaceutically acceptable salt and derivative thereof is used for inhibiting proliferation, growth, migration, infiltration, metastasis and recurrence of tumor cells, or promoting apoptosis of tumor cells.

9. The use of claim 7, wherein the tumor comprises osteosarcoma, leukemia, pancreatic cancer, lung cancer.

10. The use of claim 9, wherein the osteosarcoma cells comprise osteosarcoma cells of the SJSA-1 type, osteosarcoma cells of the MNNG/HOS type, osteosarcoma cells of the U-2-OS type, osteosarcoma cells of the MG63 type, osteosarcoma cells of the HOS type, osteosarcoma cells of the 143B type; the lung cancer cells include a549 type lung cancer cells; the leukemia includes Jurkat leukemia cells; the pancreatic cancer cells include PANC-1 pancreatic cancer cells.

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