A coumarin selenium ether thiazole hydrazone derivative, its preparation method and application

Abstract

The present invention belongs to the technical field of medicinal chemistry, and specifically relates to a coumarin selenoether thiazolylhydrazone derivative, a preparation method thereof, and an application thereof. The coumarin selenoether thiazolylhydrazone derivative has a chemical structural formula shown in Formula I. Based on the principle of drug design and splicing, the present invention designs and synthesizes a series of coumarin selenoether thiazolylhydrazone derivatives, a total of 8 coumarin selenoether thiazolylhydrazone compounds. All 8 coumarin selenoether thiazolylhydrazone compounds have good free radical scavenging ability. The coumarin selenoether thiazolylhydrazone derivative mainly binds to myeloperoxidase through π-H interaction and hydrogen bond force, and has the potential to become a myeloperoxidase inhibitor.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicinal chemistry, and particularly relates to a coumarin selenium ether thiazole hydrazone derivative, a preparation method thereof and an application thereof. Background Art

[0002] Reactive oxygen species (ROS) are generated during mitochondrial oxidative metabolism and in response to the invasion of foreign organisms, cytokines and bacteria by cells, including superoxide anion, hydroxyl radical, hydrogen peroxide, singlet molecular oxygen, etc. Under physiological conditions, the level of ROS in the body is relatively low, which is of great significance in host defense and inflammation, cell signal transduction, etc. However, when the body is subjected to damaging stimuli or in a diseased state, a large amount of ROS will be rapidly generated and accumulate in the body. When the generation amount exceeds the body's own antioxidant capacity, it will cause oxidative stress. The ROS generated during oxidative stress will cause damage to biological macromolecules such as lipids, proteins, and DNA, and then lead to various chronic and degenerative diseases, such as aging, cataract, cancer, rheumatoid arthritis, autoimmune diseases, cardiovascular and neurodegenerative diseases. The body's antioxidant defense system can counteract oxidative stress. The antioxidant defense system includes enzymatic antioxidants (endogenous antioxidants, naturally produced in the human body) and non-enzymatic antioxidants (exogenous antioxidants, obtained from the outside through food or supplements). Myeloperoxidase (MPO) is a heme peroxidase and an enzymatic antioxidant, mainly secreted by neutrophils. It can specifically catalyze the generation of the strong oxidant hypochlorous acid to scavenge harmful substances. However, during oxidative stress, MPO catalyzes the generation of excessive hypochlorous acid, which may change the structure of proteins and lipids and cause their functions to be lost. Epidemiological studies have shown that MPO is generally considered to be one of the biomarkers of inflammation and cardiovascular diseases, which has attracted the interest of medicinal chemists in designing and developing MPO inhibitors.

[0003] Only endogenous antioxidants are incomplete, and the body's antioxidant defense system also requires exogenous antioxidants. Antioxidants are becoming increasingly important as drugs for treating various lifestyle-related diseases. Coumarin, also known as 1,2-benzopyranone, is extracted and isolated from natural plants. Coumarin and its derivatives have a wide range of applications in the pharmaceutical field and possess various pharmacological activities, such as anti-cancer, anticoagulant, antiviral, anti-inflammatory, antibacterial, antifungal, antithrombotic, antioxidant, and anti-Alzheimer's. Clinically used coumarin drugs include anticoagulants warfarin and phenprocoumon, cholagogic and spasmolytic drug hymecromone, antibacterial drug novobiocin, etc. Natural compounds have low toxicity and multiple biological activities. However, due to the weak antioxidant activity and low bioavailability of natural coumarin compounds, their clinical applications are limited. Therefore, structural modification is carried out on the skeleton structure of natural product coumarin, hoping to obtain lead compounds with better antioxidant activity and bioavailability. Summary of the Invention

[0004] The purpose of the present invention is to provide a coumarin selenoether thiazolylhydrazone derivative, its preparation method and application. Based on the principle of drug design and splicing, the present invention designed and synthesized a series of coumarin selenoether thiazolylhydrazone derivatives, a total of 8 coumarin selenoether thiazolylhydrazone compounds. All 8 coumarin selenoether thiazolylhydrazone compounds have good free radical scavenging ability. The coumarin selenoether thiazolylhydrazone derivative mainly binds to myeloperoxidase through π-H interaction and hydrogen bond force and has the potential to become a myeloperoxidase inhibitor.

[0005] The present invention solves the above technical problems through the following technical solutions.

[0006] The first object of the present invention is to provide a coumarin selenoether thiazolylhydrazone derivative, which has a chemical structural formula as shown in Formula I:

[0007]

[0008] Wherein, R is selected from a substituted or unsubstituted phenyl group, and the substituents of the phenyl group are selected from C 1 -C 4 alkyl, alkoxy, hydroxyl or halogen.

[0009] Furthermore, the substituents of the phenyl group are selected from methyl, methoxy, hydroxyl, chlorine or fluorine.

[0010] Furthermore, R is selected from phenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-methylphenyl, 3-chlorophenyl or 4-fluorophenyl.

[0011] The second object of the present invention is to provide a preparation method of the above-mentioned coumarin selenoether thiazolylhydrazone derivative, which includes the following steps:

[0012] The intermediate 2a and thiosemicarbazide are dissolved in ethanol for an addition condensation reaction. After the reaction is completed, a bromoacetophenone compound is added for a Hantzch cyclization reaction. After the reaction is completed, vacuum filtration, washing, drying, and crystallization are carried out in sequence to obtain a coumarin selenoether thiazolylhydrazone compound; the synthesis reaction formula is as follows:

[0013]

[0014] Furthermore, the molar volume ratio of intermediate 2a, thiosemicarbazide, and ethanol is 0.3 mmol: 0.6 - 0.9 mmol: 5 - 10 mL. The temperature of the addition condensation reaction is 80 °C and the time is 24 h; the molar ratio of thiosemicarbazide to the bromoacetophenone compound is 1:1. The temperature of the Hantzch cyclization reaction is 80 °C and the time is 12 h.

[0015] Furthermore, the preparation method of the intermediate 2a includes the following steps:

[0016] The intermediate 1a and ethyl acetoacetate are dissolved in ethanol, and piperidine is added for a Knoevenagel condensation reaction. After the reaction is completed, vacuum filtration, washing, drying, and column chromatography separation are carried out in sequence to obtain the intermediate 2a; the synthesis reaction formula is as follows:

[0017]

[0018] Furthermore, the molar volume ratio of intermediate 1a, ethyl acetoacetate, piperidine, and ethanol is 1 mmol: 2.5 - 3 mmol: 0.1 - 0.2 mmol: 3 mL. The temperature of the Knoevenagel condensation reaction is 80 °C and the time is 1 h.

[0019] Furthermore, the preparation method of the intermediate 1a includes the following steps:

[0020] Salicylaldehyde and selenium dioxide are dissolved in hydrochloric acid for a selenization reaction. After the reaction is completed, filtration, washing, dissolution, and recrystallization are carried out in sequence to obtain the intermediate 1a; the synthesis reaction formula is as follows:

[0021]

[0022] Furthermore, the molar volume ratio of salicylaldehyde, selenium dioxide, and hydrochloric acid is 1.5 mmol: 0.5 mmol: 2 mL; the mass concentration of hydrochloric acid is 36 - 38%, the temperature of the selenization reaction is 80 °C, and the time is 50 h.

[0023] The third object of the present invention is to provide the application of the above-mentioned coumarin selenoether thiazolylhydrazone derivatives in the preparation of antioxidant drugs, and the antioxidant drugs are used to inhibit myeloperoxidase.

[0024] The present invention has the following beneficial effects compared with the prior art:

[0025] 1. Based on the principle of drug design splicing, the present invention designed and synthesized a series of coumarin selenium ether thiazolylhydrazone derivatives, a total of 8 coumarin selenium ether thiazolylhydrazone compounds; all 8 synthesized coumarin selenium ether thiazolylhydrazone compounds have good free radical scavenging ability, and among them, compounds 3a, 3e and 3f have stronger free radical scavenging ability against three kinds of free radicals than the positive control L-ascorbic acid.

[0026] 2. The coumarin selenium ether thiazolylhydrazone derivatives synthesized in the present invention mainly bind to myeloperoxidase through pi-H interaction and hydrogen bond force, and have the potential to become myeloperoxidase inhibitors. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1-2 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 1a of the present invention;

[0028] Figure 3-4 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 2a of the present invention;

[0029] Figure 5-6 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3a of the present invention;

[0030] Figure 7-8 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3b of the present invention;

[0031] Figure 9-10 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3c of the present invention;

[0032] Figure 11-12 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3d of the present invention;

[0033] Figure 13-14 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3e of the present invention;

[0034] Figure 15-16 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3f of the present invention;

[0035] Figure 17-18 They are respectively the 1H NMR spectrum and 13C NMR spectrum of compound 3g of the present invention;

[0036] Figure 19-21 They are respectively the 1H NMR spectrum, 13C NMR spectrum and 19F NMR spectrum of compound 3h of the present invention;

[0037] Figure 22-24The DPPH· scavenging rate, Galvinoxyl· scavenging rate, and ABTS + · scavenging rate graphs of the coumarin selenium ether thiazolylhydrazone compounds prepared in Examples 3-10 of the present invention;

[0038] Figure 25 Figure showing the binding mode of Compound 3e of the present invention to the active site of myeloperoxidase (MPO). Detailed implementation manners

[0039] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention.

[0040] It should be noted that the professional terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the protection scope of the present invention. Unless otherwise specifically stated, various raw materials, reagents, instruments, and equipment used in the following embodiments of the present invention can be obtained through market purchase or prepared by existing methods. The salicylaldehyde, selenium dioxide, thiosemicarbazide, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-di-tert-butyl-α-(3,5-di-tert-butyl-4-oxo-2,5-cyclohexadienylidene)-p-tolyloxy (Galvinoxyl), and bromoacetophenone compounds used in the present invention are purchased from Shanghai Titan Technology Co., Ltd.; ethyl acetoacetate is purchased from Xilong Scientific Co., Ltd.; all the reagents used are of analytical grade.

[0041] Example 1

[0042] Preparation method of diaryl selenium compound (Intermediate 1a), comprising the following steps:

[0043] Take salicylaldehyde (0.1832 g, 1.5 mmol), selenium dioxide (0.0555 g, 0.5 mmol), and 2 mL of concentrated hydrochloric acid with a mass concentration of 36-38%, add them to a 15 mL round-bottom flask equipped with a magnetic stirrer, stir at room temperature for 0.5 h, then place the reaction flask in an oil bath at 80 °C and heat under reflux. After reacting for 50 h, a large amount of yellow solid is produced; filter the produced yellow solid, wash it 3 times with deionized water, dry it, dissolve the dried crude product by stirring and heating with ethyl acetate and absolute ethanol (volume ratio 5:1), recrystallize three times, and finally obtain a yellow crystal compound, which is diaryl selenium compound 1a, with a yield of 39%; mp 155.4-156.3 °C; the synthesis reaction formula is as follows:

[0044]

[0045] 1 1H NMR (400 MHz, DMSO-d 6 ): δ 10.95 (s, 2H), 10.21 (s, 2H), 7.70 (d, J = 2.4 Hz, 2H), 7.59 (dd, J = 8.6, 2.4 Hz, 2H), 6.98 (d, J = 8.6 Hz, 2H); as Figure 1 shown; 13 13C NMR (100 MHz, DMSO-d 6 ): δ 190.85, 161.05, 141.39, 133.68, 123.75, 120.21, 119.44; as Figure 2 shown; HRMS (ESI) calcd for C 14 H 10 O 4 Se [M+H] + 322.9744, found 322.9817.

[0046] Example 2

[0047] Preparation method of intermediate 2a, comprising the following steps:

[0048] Take the diarylselenium compound of Example 1 (0.3212 g, 1 mmol), ethyl acetoacetate (380 μL, 0.3904 g, 3 mmol) and 3 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stir bar, place it in an 80 °C heating plate and heat for 10 min, then add piperidine (10 μL, 0.0085 g, 0.1 mmol), the solution turns blood red, monitor the reaction process by TLC, the reaction is complete in about 1 h, a large amount of yellow solid precipitates, cool to room temperature, filter off the reaction solution under reduced pressure, wash with absolute ethanol 3 times, dry, then dissolve the dried crude product in dichloromethane, add 100-mesh silica gel, spin-dry to make sand, perform column chromatography separation on it using a dichloromethane and ethyl acetate (volume ratio 50:1) system, spin-dry to obtain a yellow solid, and obtain intermediate 2a with a yield of 83%; the synthesis reaction formula is as follows:

[0049]

[0050] 1 1H NMR (400 MHz, Chloroform-d): δ 8.39 (d, J = 0.7 Hz, 2H), 7.75 - 7.69 (m, 4H), 7.31 (d, J = 8.3 Hz, 2H), 2.71 (s, 6H); as Figure 3 shown;13 C NMR (100 MHz, Chloroform-d): δ 195.10, 158.64, 154.94, 146.40, 138.85, 134.39, 126.61, 125.25, 119.42, 118.07, 30.54; as Figure 4 shown; HRMS (ESI) calcd for C 22 H 14 O 6 Se [M+Na] + 470.9907, found 470.9918.

[0051] Example 3

[0052] A preparation method of 6,6'-selenobis(3-(1-(2-(4-phenylthiazol-2-yl)hydrazinylidene)ethyl)-2H-benzopyran-2-one) (3a) comprises the following steps:

[0053] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them into a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and dropwise add 2 drops of acetic acid. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromoacetophenone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash it 3 times with deionized water and absolute ethanol respectively, dry it, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3a; yellowish-brown solid, yield 73%; the structural formula of compound 3a is as follows:

[0054]

[0055] 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.40 (s, 2H), 8.12 (s, 2H), 8.03 (d, J = 2.1 Hz, 2H), 7.86 (d, J = 7.7 Hz, 4H), 7.69 (dd, J = 8.5, 2.1 Hz, 2H), 7.39 (dd, J = 8.3, 4.7 Hz, 6H), 7.30 (d, J = 7.5 Hz, 4H), 2.26 (s, 6H); as Figure 5 shown; 13 C NMR (100 MHz, DMSO-d 6): δ 169.70, 159.29, 153.38, 151.19, 145.10, 140.35, 136.83, 135.13, 133.66, 129.08, 128.00, 127.64, 126.17, 125.99, 120.67, 117.89, 104.85, 16.64; as Figure 6 shown; HRMS(ESI) calcd for C 40 H 28 N 6 O 4 S 2 Se[M + H] + 795.0911, found 795.0967.

[0056] Example 4

[0057] Preparation method of 6,6'-selenobis(3-(-1-(2-(4-(3-chlorophenyl)thiazol-2-yl)hydrazono)ethyl)-2H-benzopyran-2-one (3b), comprising the following steps:

[0058] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and dropwise add 2 drops of acetic acid. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(3-chlorophenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash it 3 times with deionized water and absolute ethanol respectively, dry it, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3b; yellowish-brown solid, yield 77%; the structural formula of compound 3b is as follows:

[0059]

[0060] 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.38 (s, 2H), 8.07 (s, 2H), 7.95 (s, 2H), 7.88 (s, 2H), 7.78 (d, J = 7.8 Hz, 2H), 7.64 (d, J = 8.6 Hz, 2H), 7.44 - 7.28 (m, 8H), 2.23 (s, 6H); as Figure 7 shown; 13 C NMR (100 MHz, DMSO-d 6)): δ 169.77, 159.24, 153.32, 149.48, 145.24, 140.35, 137.08, 136.82, 133.96, 133.58, 130.96, 127.68, 127.45, 126.16, 125.70, 124.43, 120.57, 117.85, 106.41, 16.59; as Figure 8 shown; HRMS(ESI) calcd for C 40 H 26 Cl 2 N 6 O 4 S 2 Se[M + H] + 863.0132, found 863.0222.

[0061] Example 5

[0062] Preparation method of 6,6'-selenobis(3-(1-(2-(4-(3-methoxyphenyl)thiazol-2-yl)hydrazono)ethyl)-2H-chromen-2-one) (3c), comprising the following steps:

[0063] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and add 2 drops of acetic acid. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(3-methoxyphenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash 3 times with deionized water and absolute ethanol respectively, dry, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3c; yellowish-brown solid, yield 59%; the structural formula of compound 3c is as follows:

[0064]

[0065] 1 H NMR (400 MHz, DMSO-d 6): δ 11.34 (s, 2H), 8.07 (s, 2H), 7.96 (d, J = 2.2 Hz, 2H), 7.64 (dd, J = 8.6, 2.1 Hz, 2H), 7.42 (s, 2H), 7.34 (d, J = 8.6 Hz, 2H), 7.29 (d, J = 7.4 Hz, 4H), 6.88 - 6.81 (m, 4H), 3.80 (d, J = 1.9 Hz, 0H), 3.77 (s, 6H), 2.24 (s, 6H); as Figure 9 shown; 13 C NMR (100 MHz, DMSO - d 6 ): δ 169.60, 159.98, 159.27, 153.31, 150.82, 145.11, 140.28, 136.79, 136.45, 133.56, 130.12, 127.52, 126.16, 120.59, 118.37, 117.83, 113.91, 111.16, 105.17, 55.53, 16.60; as Figure 10 shown; HRMS (ESI) calcd for C 42 H 32 N 6 O 6 S 2 Se[M + H] + 855.1122, found 855.1091.

[0066] Example 6

[0067] Preparation method of 6,6'-selenobis(3-(1-(2-(4-(4 - methoxyphenyl)thiazol - 2 - yl)hydrazono)ethyl)-2H - chromen - 2 - one) (3d), comprising the following steps:

[0068] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them into a 15 mL high - pressure reaction tube equipped with a magnetic stirrer, and add 2 drops of acetic acid. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2 - bromo - 1-(4 - methoxyphenyl)ethanone into the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash it 3 times with deionized water and absolute ethanol respectively, dry it, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin - selenium ether - thiazolylhydrazone derivative 3d; yellow - brown solid, yield 64%; the structural formula of compound 3d is as follows:

[0069]

[0070] 1 1H NMR (400 MHz, DMSO-d 6 ): δ 11.35 (s, 2H), 8.10 (s, 2H), 8.01 (d, J = 2.1 Hz, 2H), 7.81 - 7.73 (m, 4H), 7.67 (dd, J = 8.6, 2.1 Hz, 2H), 7.38 (d, J = 8.6 Hz, 2H), 7.11 (s, 2H), 6.99 - 6.91 (m, 4H), 3.77 (s, 6H), 2.25 (s, 6H); as Figure 11 shown; 13 13C NMR (100 MHz, DMSO-d 6 ): δ 169.59, 159.31, 159.24, 153.35, 144.95, 140.28, 136.81, 133.63, 129.52, 127.95, 127.63, 127.32, 126.17, 120.66, 117.87, 114.42, 102.63, 55.57, 16.61; as Figure 12 shown; HRMS (ESI) calcd for C 42 H 32 N 6 O 6 S 2 Se [M + H] + 855.1122, found 855.1164.

[0071] Example 7

[0072] Preparation method of 6,6'-selenobis(3-(1-(2-(4-(2-hydroxyphenyl)thiazol-2-yl)hydrazono)ethyl)-2H-chromen-2-one) (3e), comprising the following steps:

[0073] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and add 2 drops of acetic acid dropwise. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(2-hydroxyphenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash 3 times with deionized water and absolute ethanol respectively, dry, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3e; yellowish-brown solid, yield 64%; the structural formula of compound 3e is as follows:

[0074]

[0075] 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.39 (d, J = 73.8 Hz, 2H), 8.12 (s, 2H), 8.00 (d, J = 2.1 Hz, 2H), 7.79 (dd, J = 7.8, 1.7 Hz, 2H), 7.67 (dd, J = 8.6, 2.1 Hz, 2H), 7.38 (d, J = 8.1 Hz, 4H), 7.15 (s, 2H), 6.90 - 6.81 (m, 6H), 2.27 (s, 6H); as Figure 13 shown; 13 C NMR (100 MHz, DMSO-d 6 ): δ 168.92, 161.47, 159.24, 155.67, 153.38, 148.46, 145.75, 140.51, 136.90, 133.66, 129.48, 127.42, 126.20, 120.60, 119.55, 119.21, 117.89, 117.34, 104.95, 16.72; as Figure 14 shown; HRMS (ESI) calcd for C 40 H 28 N 6 O 6 S 2 Se [M + H] + 827.0809, found 827.1199

[0076] Example 8

[0077] A method for preparing 6,6'-selenobis(3-(1-(2-(4-(3-hydroxyphenyl)thiazol-2-yl)hydrazono)ethyl)-2H-chromen-2-one) (3f) comprises the following steps:

[0078] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and add 2 drops of acetic acid dropwise. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(3-hydroxyphenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash it 3 times with deionized water and absolute ethanol respectively, dry it, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3f; brownish-yellow solid, yield 56%; the structural formula of compound 3f is as follows:

[0079]

[0080] 1 H NMR(400MHz,DMSO-d 6 ):δ11.35(s,2H),9.47(s,2H),8.11(s,2H),8.01(d,J=2.1Hz,2H),7.68(dd,J=8.6,2.1Hz,2H),7.39(d,J=8.6Hz,2H),7.29-7.26(m,4H),7.18(t,J=7.9Hz,4H),6.70(d,J=6.7Hz,2H),2.25(s,6H); as Figure 15 shown; 13 C NMR(100MHz,DMSO-d 6 ):δ169.52,159.31,158.01,153.36,151.28,145.13,140.34,136.84,136.44,133.64,130.03,127.65,126.18,120.66,117.89,116.95,115.11,113.00,104.72,16.63; as Figure 16 shown; HRMS(ESI)calcd for C 40 H 28 N 6 O 6 S 2 Se[M+Na] + 849.0629,found 849.0694.

[0081] Example 9

[0082] Preparation method of 6,6'-selenobis(3-(1-(2-(4-(4-methylphenyl)thiazol-2-yl)hydrazinylidene)ethyl)-2H-chromen-2-one) (3g), comprising the following steps:

[0083] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stirrer, and add 2 drops of acetic acid dropwise. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(4-methylphenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash with deionized water and absolute ethanol three times respectively, dry, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3g; yellowish-brown solid, yield 71%; the structural formula of compound 3g is as follows:

[0084]

[0085] 1 H NMR (400 MHz, DMSO-d 6 ): δ11.37 (s, 2H), 8.12 (s, 2H), 8.03 (d, J = 2.2 Hz, 2H), 7.76 - 7.67 (m, 6H), 7.39 (d, J = 8.6 Hz, 2H), 7.24 - 7.17 (m, 6H), 2.31 (s, 6H), 2.25 (s, 6H); as Figure 17 shown; 13 C NMR (100 MHz, DMSO-d 6 ): δ169.60, 159.31, 153.37, 151.16, 145.00, 140.32, 137.27, 136.83, 133.66, 132.45, 129.65, 127.65, 126.17, 125.94, 120.68, 117.89, 103.90, 21.27, 16.62; as Figure 18 shown; HRMS (ESI) calcd for C 42 H 32 N 6 O 4 S 2 Se[M + H] + 823.1224, found 823.1266.

[0086] Example 10

[0087] Preparation method of 6,6'-selenobis(3-(1-(2-(4-(4-fluorophenyl)thiazol-2-yl)hydrazinylidene)ethyl)-2H-chromen-2-one) (3h), comprising the following steps:

[0088] Take the intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of absolute ethanol, add them to a 15 mL high-pressure reaction tube equipped with a magnetic stir bar, and add 2 drops of acetic acid. After heating in an 80 °C heating plate for 24 h, directly add 0.63 mmol of 2-bromo-1-(4-fluorophenyl)ethanone to the reaction tube. Monitor the reaction process by TLC. The reaction is complete in about 12 h. Cool to room temperature, filter off the reaction solution under reduced pressure, wash 3 times with deionized water and absolute ethanol respectively, dry, and then crystallize the dried crude product with pyridine and absolute ethanol (volume ratio 1:10) to obtain the coumarin selenium ether thiazolylhydrazone derivative 3h; yellowish-brown solid, yield 78%; the structural formula of compound 3h is as follows:

[0089]

[0090] 1 H NMR(400MHz,DMSO-d 6 ):δ11.39(s,2H),8.11(s,2H),8.01(d,J=2.1Hz,2H),7.90-7.85(m,4H),7.67(dd,J=8.6,2.1Hz,2H),7.38(d,J=8.6Hz,2H),7.23(d,J=8.9Hz,6H),2.25(s,6H); as Figure 19 shown; 13 C NMR(100MHz,DMSO-d 6 ):δ169.79,163.27,160.84,159.28,153.36,145.22,140.35,136.84,133.63,127.95(d,J=8.1Hz),127.59,126.16,120.65,117.88,116.02,115.80,104.58,16.63; as Figure 20 shown; 19 F NMR(376MHz,DMSO-d 6 ):δ-114.56; as Figure 21 shown; HRMS(ESI)calcd for C 40 H 26 F 2 N 6 O 4 S 2Se[M+H] + 831.0723, found 831.1199.

[0091] Example 11

[0092] The coumarin selenium ether thiazole hydrazone compounds prepared in Examples 3-10 were subjected to antioxidant activity determination

[0093] 1. Determination of DPPH radical scavenging rate

[0094] Add 100 μL of DPPH ethanol solution (25 μg / mL) to a 96-well microplate, and then add 100 μL of sample solutions with different concentrations (200, 100, 50, 25, 12.5, 6.25 μmol / L) to each well. React at room temperature in the dark for 30 min, and measure the absorbance value A of the sample group at a wavelength of 517 nm using a microplate reader 1 , with 3 parallels in each group, and L-ascorbic acid as the positive control.

[0095] DPPH radical scavenging rate = [(A 3 - A 0 ) - (A 1 - A 2 )] / (A 3 - A 0 ) * 100%; The absorbance value A 2 of the sample blank group is 100 μL of sample solution + 100 μL of ethanol solvent, and the absorbance value A 3 of the negative control group is 100 μL of DPPH ethanol solution + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio), and the absorbance A 0 of the blank group is 100 μL of ethanol + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio).

[0096] 2. Determination of Galvinoxyl radical scavenging rate

[0097] Add 100 μL of Galvinoxyl ethanol solution (30 μg / mL) to a 96-well microplate, and then add 100 μL of sample solutions with different concentrations (50, 40, 30, 20, 10, 5 μmol / L) to each well. React at room temperature in the dark for 30 min, and measure the absorbance value A of the sample group at a wavelength of 428 nm using a microplate reader 1 , with 3 parallels in each group, and L-ascorbic acid as the positive control.

[0098] Galvinoxyl radical scavenging rate = [(A 3 - A 0 ) - (A 1 - A2 )] / (A 3 -A 0 ) * 100%; The absorbance value A of the sample blank group 2 is 100 μL of sample solution + 100 μL of ethanol, and the absorbance value A of the negative control group 3 is 100 μL of Galvinoxyl ethanol solution + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio), and the absorbance A of the solvent blank group 0 is 100 μL of ethanol + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio).

[0099] 3. Determination of ABTS cation radical scavenging

[0100] Take 10 mg of ABTS and 3 mg of K 2 S 2 O 8 Add them to a 10 mL volumetric flask, dilute with deionized water to the mark, and let it stand in the dark at room temperature for 24 h to prepare the ABTS working solution. After the oxidation is completed, dilute with absolute ethanol to make the absorbance of the ABTS solution at 734 nm be (0.700 ± 0.020). Add 100 μL of the ABTS working solution to a 96-well microplate, and then add 100 μL of sample solutions with different concentrations (80, 60, 40, 20, 10, 5 μmol / L) to each well respectively. React at room temperature in the dark for 12 min, and use a microplate reader to measure the absorbance value A of the sample group 1 , with 3 parallels set for each group, and L-ascorbic acid as the positive control.

[0101] ABTS + radical scavenging rate = [(A 3 -A 0 ) - (A 1 -A 2 )] / (A 3 -A 0 ) * 100%; The absorbance value A of the sample blank group 2 is 100 μL of sample solution + 100 μL of ethanol, and the absorbance value A of the negative control group 3 is 100 μL of ABTS ethanol solution + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio), and the absorbance A of the solvent blank group 0 is 100 μL of ethanol + 100 μL of a mixed solvent (DMSO:EtOH = 1:5, volume ratio).

[0102] DPPH·, Galvinoxyl·, ABTS +·They show purple, yellow, and blue-green colors respectively in ethanol solution, and have maximum absorption at 517 nm, 428 nm, and 734 nm respectively. When free radicals react with compounds with antioxidant effects, the absorbance value will decrease.

[0103] Figure 22 This is the DPPH· scavenging rate of the coumarin selenium ether thiazolylhydrazone compounds prepared in Examples 3-10 of the present invention. As Figure 22 shown, all 8 compounds 3a-3h have the ability to scavenge DPPH free radicals; except for 3b and 3h, the IC 50 values of the remaining compounds are lower than the IC 50 value of L-ascorbic acid, 12.56 μmol / L, indicating that their antioxidant effect is stronger than that of L-ascorbic acid. Among them, compound 3e has the strongest activity, and its IC 50 value for scavenging DPPH free radicals is 7.47 μmol / L; the IC 50 in Table 1 shows that the compounds with methoxy and phenolic hydroxyl substitutions (3c, 3d, 3e, 3f) have stronger DPPH free radical scavenging ability than the compounds substituted with methyl and halogen atoms (3b, 3g, 3h). It is speculated that compounds containing electron-donating groups may have stronger DPPH free radical scavenging ability than compounds containing electron-withdrawing groups.

[0104] Figure 23 This is the Galvinoxyl· scavenging rate of the coumarin selenium ether thiazolylhydrazone compounds prepared in Examples 3-10 of the present invention. As Figure 23 shown, except for compound 3d, the scavenging rates of the remaining compounds for Galvinoxyl free radicals at 25 μmol / L are stronger than that of L-ascorbic acid. It can be seen from Table 1 that the IC 50 values of compounds 3a-3h are 4.62-11.16 μmol / L, which are similar to the IC 50 value of L-ascorbic acid, 8.56 μmol / L, indicating that their antioxidant effect is comparable to that of L-ascorbic acid. Among them, compound 3e has the strongest activity, and its IC 50 value for scavenging Galvinoxyl free radicals is 4.62 μmol / L.

[0105] Figure 24 This is the ABTS + · scavenging rate of the coumarin selenium ether thiazolylhydrazone compounds prepared in Examples 3-10 of the present invention. As Figure 24 shown, all 8 compounds 3a-3h have the ability to scavenge ABTS cationic free radicals. Among them, the scavenging rates of compounds 3e, 3f, and 3g for ABTS cationic free radicals at 40 μmol / L are stronger than that of L-ascorbic acid; it can be seen from Table 1 that the IC 50The value is 7.70 - 29.71 μmol / L. Among them, the IC50 values of compounds 3a, 3e, 3f, and 3g are all lower than the IC 50 value of 13.94 μmol / L of L-ascorbic acid, indicating that the antioxidant effects of compounds 3a, 3e, 3f, and 3g are all stronger than that of L-ascorbic acid.

[0106] Table 1 Antioxidant activities of compounds 3a - 3h

[0107]

[0108]

[0109] Example 11

[0110] Retrieve and download myeloperoxidase (encoded by 4C1M) as the receptor protein through the Protein Data Bank at https: / / www.rcsb.org / . Use the MOE software to perform preprocessing on the downloaded protein, including deleting duplicate chains, deleting water molecules, adding hydrogen atoms, and energy minimization, to establish a small molecule database of coumarin selenoether thiazolylhydrazone compounds. Through the Compute-Dock operation, dock the compounds in the small molecule database with the processed protein. The smaller the docking score, the less energy required for docking, and the more spontaneously the reaction can proceed. After docking, through the Ligand Interactions operation, information such as whether the docked compound has a side chain exposed, the binding mode of the compound to the key amino acids in the active center, and the interaction force between the compound and the enzyme can be visually displayed.

[0111] Use the MOE software to perform molecular docking of compounds 3a - 3h with myeloperoxidase, and preliminarily explore their interactions and the best docking conformations. Taking compound 3e as an example, Figure 25 This is the binding mode diagram of compound 3e of the present invention with the active site of myeloperoxidase (MPO). As Figure 25 shown, the interaction between compound 3e and the residues of myeloperoxidase can be observed. Their docking score is -7.6489 (Table 2). A total of 5 amino acids participate in the interaction. Compound 3e forms hydrogen bonds with Ser266 and Asn186 in the form of H-donor through phenolic hydroxyl groups, forms a pi-H interaction with Asn189 through the thiazole ring, and forms pi-H interactions with Asn192 and Gln193 through the benzene ring on the coumarin parent nucleus. The docking score is negative, and the smaller the score, the closer the compound binds to the protein. Table 2 lists the docking results of 3a - 3h, indicating that these 8 compounds theoretically all have good affinity with myeloperoxidase and all have the potential to be myeloperoxidase inhibitors.

[0112] Table 2 Docking effects of compounds 3a - 3h with myeloperoxidase

[0113]

[0114]

[0115] In summary, eight compounds of coumarin selenium ether thiazole hydrazone derivatives were synthesized in this invention, and the in vitro antioxidant properties of coumarin selenium ether thiazole hydrazone derivatives and their molecular mechanism of action on myeloperoxidase inhibition were investigated. The results of in vitro antioxidant assay showed that in three test systems, the eight coumarin selenium ether thiazole hydrazone derivatives all had good free radical scavenging ability, indicating that these compounds are potential antioxidants; the results of molecular docking showed that coumarin selenium ether thiazole hydrazone derivatives mainly bind to myeloperoxidase through π-H interaction and hydrogen bond force, and have the potential to become myeloperoxidase inhibitors.

[0116] It should be noted that when the present invention involves a numerical range, it should be understood that any value between the two endpoints of each numerical range and the two endpoints can be selected. Since the steps and methods adopted are the same as those in the examples, in order to avoid repetition, the present invention describes the preferred embodiments. Although the preferred embodiments of the present invention have been described, those skilled in the art can make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the appended claims are intended to be construed as including the preferred embodiments and all changes and modifications falling within the scope of the present invention.

[0117] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. A coumarin selenium ether thiazole hydrazone derivative, characterized in that, it has a chemical structural formula shown in Formula I: Among them, R is selected from substituted or unsubstituted phenyl, and the substituents of phenyl are selected from C 1 -C 4 alkyl, alkoxy, hydroxy or halogen.

2. The coumarin selenium ether thiazole hydrazone derivative according to claim 1, characterized in that, the substituents of the phenyl group are selected from methyl, methoxy, hydroxy, chloro or fluoro.

3. The coumarin selenium ether thiazole hydrazone derivative according to claim 1, characterized in that, R is selected from phenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-methylphenyl, 3-chlorophenyl or 4-fluorophenyl.

4. A preparation method of the coumarin selenium ether thiazole hydrazone derivative according to any one of claims 1-3, characterized in that, it includes the following steps: Dissolve intermediate 2a and thiosemicarbazide in ethanol for addition condensation reaction. After the reaction is completed, add bromoacetophenone compounds for Hantzch cyclization reaction. After the reaction is completed, carry out vacuum filtration, washing, drying and crystallization in sequence to obtain the coumarin selenium ether thiazole hydrazone compound; The synthesis reaction formula is shown as follows:

5. The preparation method of the coumarin selenium ether thiazole hydrazone derivative according to claim 4, characterized in that, The molar volume ratio of intermediate 2a, thiosemicarbazide and ethanol is 0.3 mmol: 0.6 ～ 0.9 mmol: 5 ～ 10 mL. The temperature of the addition condensation reaction is 80 °C and the time is 24 h; the molar ratio of thiosemicarbazide to bromoacetophenone compounds is 1:1, and the temperature of the Hantzch cyclization reaction is 80 °C and the time is 12 h.

6. The preparation method of the coumarin selenium ether thiazole hydrazone derivative according to claim 4, characterized in that, The preparation method of the intermediate 2a includes the following steps: Dissolve intermediate 1a and ethyl acetoacetate in ethanol, add piperidine and then carry out Knoevenagel condensation reaction. After the reaction is completed, carry out vacuum filtration, washing, drying and column chromatography separation in sequence to obtain intermediate 2a; The synthesis reaction formula is shown as follows:

7. The preparation method of the coumarin selenium ether thiazole hydrazone derivative according to claim 6, characterized in that, The molar volume ratio of intermediate 1a, ethyl acetoacetate, piperidine and ethanol is 1 mmol: 2.5 ～ 4 mmol: 0.1 ～ 0.2 mmol: 3 mL. The temperature of the Knoevenagel condensation reaction is 80 °C and the time is 1 h.

8. The preparation method of the coumarin selenium ether thiazole hydrazone derivative according to claim 6, characterized in that, The preparation method of the intermediate 1a includes the following steps: Carry out selenization reaction by dissolving salicylaldehyde and selenium dioxide in hydrochloric acid. After the reaction is completed, filter, wash, dissolve and recrystallize in sequence to obtain intermediate 1a; The synthesis reaction formula is shown as follows:

9. The preparation method of the coumarin selenium ether thiazole hydrazone derivative according to claim 8, characterized in that, the molar volume ratio of salicylaldehyde, selenium dioxide and hydrochloric acid is 1.5 mmol: 0.5 mmol: 2 mL; The mass concentration of hydrochloric acid is 36 ～ -38%, the temperature of the selenization reaction is 80 °C, and the time is 50 h.

10. An application of the coumarin selenium ether thiazole hydrazone derivative according to claim 1 in the preparation of antioxidant drugs, characterized in that, the antioxidant drug is used to scavenge free radicals, and the free radicals are at least one of DPPH free radicals, Galvinoxyl free radicals and ABTS cation free radicals.

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