Coumarin selenide thiazole hydrazone derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a coumarin selenide thiazole hydrazone derivative as well as a preparation method and application thereof. The coumarin selenide thiazole hydrazone derivative has a chemical structural formula as shown in a formula I which is described in the specification. A series of coumarin selenide thiazole hydrazone derivatives are designed and synthesized on the basis of the drug design splicing principle, the total number of the coumarin selenide thiazole hydrazone derivatives is eight, and the eight coumarin selenide thiazole hydrazone compounds all have good free radical scavenging capacity and can be used for preparing the coumarin selenide thiazole hydrazone derivatives. The coumarin selenide thiazole hydrazone derivative is mainly combined with myeloperoxidase through pi-H action and hydrogen-bond interaction, and has the potential of becoming a myeloperoxidase inhibitor. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to a coumarin selenoether thiazole hydrazone derivative and a preparation method and application thereof. Background Art

[0002] Reactive oxygen species (ROS) are produced during mitochondrial oxidative metabolism and in the cell's response to foreign organisms, cytokines and bacterial invasion, including superoxide anions, hydroxyl radicals, hydrogen peroxide, singlet molecular oxygen, etc. Under physiological conditions, the level of ROS in the body is low, which is of great significance in host defense, inflammation, cell signaling, etc. However, when the body is subjected to damaging stimulation or in a disease state, a large amount of ROS will be rapidly generated and accumulated in the body. When the amount generated exceeds the body's own antioxidant capacity, it will cause oxidative stress. The ROS produced 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, cataracts, cancer, rheumatoid arthritis, autoimmune diseases, cardiovascular and neurodegenerative diseases. The human body's antioxidant defense system can fight oxidative stress. The antioxidant defense system includes enzyme 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, an enzyme antioxidant, mainly secreted by neutrophils. It can specifically catalyze the production of hypochlorous acid, a strong oxidant, to remove harmful substances. However, under oxidative stress, MPO catalyzes the production of excessive hypochlorous acid, which may change the structure of proteins and lipids, leading to loss of their function. Epidemiological studies have shown that MPO is generally considered to be one of the biomarkers of inflammation and cardiovascular disease, which has aroused the interest of medicinal chemists in designing and developing MPO inhibitors.

[0003] Endogenous antioxidants alone 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-benzopyrone, is extracted and separated from natural plants. Coumarin and its derivatives are widely used in the medical field and have a variety of pharmacological activities, such as anticancer, anticoagulant, antiviral, anti-inflammatory, antibacterial, antifungal, antithrombotic, antioxidant, and anti-Alzheimer's disease. Coumarin drugs that have been used clinically include anticoagulants warfarin and phenprocoumon, choleretic and antispasmodic drugs hydroxymethylcoumarin, and antibacterial drugs neomycin. Natural compounds have low toxicity and multiple biological activities. Due to the defects of weak antioxidant activity and low bioavailability of natural coumarin compounds, their clinical application is limited. Therefore, structural modification is performed on the skeleton structure of natural product coumarin, hoping to obtain lead compounds with good antioxidant activity and bioavailability. Summary of the invention

[0004] The purpose of the present invention is to provide a coumarin selenoether thiazole hydrazone derivative and a preparation method and application thereof. The present invention is based on the principle of drug design splicing, and designs and synthesizes a series of coumarin selenoether thiazole hydrazone derivatives, including 8 coumarin selenoether thiazole hydrazone compounds. All of the 8 coumarin selenoether thiazole hydrazone compounds have good free radical scavenging ability. The coumarin selenoether thiazole hydrazone derivatives mainly bind to myeloperoxidase through pi-H interaction and hydrogen bonding force, and have the potential to become myeloperoxidase inhibitors.

[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 thiazole hydrazone derivative having a chemical structure as shown in Formula I:

[0007]

[0008] Wherein, R is selected from substituted or unsubstituted phenyl, and the substituent of phenyl is selected from C1-C4 alkyl, alkoxy, hydroxyl or halogen.

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

[0010] Further, 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 method for preparing the above-mentioned coumarin selenoether thiazole hydrazone derivatives, comprising the following steps:

[0012] The intermediate 2a and thiosemicarbazide are dissolved in ethanol for addition condensation reaction. After the reaction, a bromoacetophenone compound is added to carry out a Hantzch cyclization reaction. After the reaction, vacuum filtration, washing, drying and crystallization are carried out in sequence to obtain a coumarin selenoether thiazole hydrazone compound. The synthetic reaction formula is shown below:

[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 and bromoacetophenone compounds is 1:1, and 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 comprises the following steps:

[0016] The intermediate 1a and ethyl acetoacetate are dissolved in ethanol, and piperidine is added to carry out Knoevenagel condensation reaction. After the reaction is completed, vacuum filtration, washing, drying, and chromatography separation are carried out in sequence to obtain the intermediate 2a. The synthetic reaction formula is shown below:

[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, and 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 comprises the following steps:

[0020] Salicylaldehyde and selenium dioxide are dissolved in hydrochloric acid for selenization reaction. After the reaction, the intermediate 1a is obtained by filtering, washing, dissolving and recrystallizing in sequence. The synthetic reaction formula is shown below:

[0021]

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

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

[0024] Compared with the prior art, the present invention has the following beneficial effects:

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

[0026] 2. The synthetic coumarin selenoether thiazole hydrazone derivatives of the present invention bind to myeloperoxidase mainly through pi-H interaction and hydrogen bonding force, and have the potential to become myeloperoxidase inhibitors. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

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

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

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

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

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

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

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

[0037] Figure 22-24The DPPH·scavenging rate, Galvinoxyl·scavenging rate and ABTS·scavenging rate of the coumarin selenoether thiazole hydrazone compounds prepared in Examples 3-10 of the present invention are respectively + Clearance graph;

[0038] Fig.25 The diagram shows the binding mode of compound 3e of the present invention to the active site of myeloperoxidase (MPO). DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within 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 scope of protection of the present invention. Unless otherwise specified, the various raw materials, reagents, instruments and equipment used in the following embodiments of the present invention can be purchased from the market or prepared by existing methods. The salicylaldehyde, selenium dioxide, semicarbazide, 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH), 2,2'-azobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,6-di-tert-butyl-α-(3,5-di-tert-butyl-4-oxygen-2,5-cyclohexadienyl)-p-tolyloxy (Galvinoxyl) and bromoacetophenone compounds used in the present invention were purchased from Shanghai Titan Technology Co., Ltd.; ethyl acetoacetate was purchased from Xilong Science Co., Ltd.; all reagents used were analytically pure.

[0041] Example 1

[0042] The preparation method of the diaryl selenium compound (intermediate 1a) comprises the following steps:

[0043] 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% were added to a 15 mL round-bottom flask equipped with a magnetic stirrer, stirred at room temperature for 0.5 h, and then the reaction flask was placed in an oil bath at 80°C and heated to reflux. After reacting for 50 h, a large amount of yellow solid was produced; the produced yellow solid was filtered, washed with deionized water 3 times, and dried. The dried crude product was stirred and heated with ethyl acetate and anhydrous ethanol (volume ratio of 5:1) to dissolve, and recrystallized three times to obtain a yellow crystalline compound, namely diaryl selenium compound 1a, with a yield of 39%; mp 155.4-156.3°C; the synthetic reaction formula is as follows:

[0044]

[0045] 1 H NMR (400MHz, DMSO-d6): δ10.95 (s, 2H), 10.21 (s, 2H), 7.70 (d, J = 2.4Hz, 2H), 7.59 (dd, J = 8.6, 2.4Hz, 2H), 6.98 (d, J = 8.6Hz, 2H); such as Figure 1 As shown; 13 CNMR (100MHz, DMSO-d6): δ190.85, 161.05, 141.39, 133.68, 123.75, 120.21, 119.44; Figure 2 HRMS (ESI) calculation for C 14 H 10 O4Se[M+H] + 322.9744, found 322.9817.

[0046] Example 2

[0047] The preparation method of intermediate 2a comprises the following steps:

[0048] Take the diaryl selenium compound (0.3212 g, 1 mmol) of Example 1, ethyl acetoacetate (380 μL, 0.3904 g, 3 mmol) and 3 mL of anhydrous ethanol, add them into a 15 mL high pressure reaction tube equipped with a magnetic stirrer, put it into an 80 ° C heating plate and heat it for 10 min, then add piperidine (10 μL, 0.0085 g, 0.1 mmol), the solution turns blood red, TLC monitors the reaction progress, the reaction is complete in about 1 h, a large amount of yellow solid is precipitated, and the reaction solution is cooled to room temperature, and the reaction solution is filtered off under reduced pressure, washed with anhydrous ethanol 3 times, and dried. The dried crude product is then dissolved in dichloromethane, 100 mesh silica gel is added, and the sand is spin-dried. The system of dichloromethane and ethyl acetate (volume ratio is 50:1) is used for column chromatography separation, and a yellow solid is obtained after spin-drying to obtain intermediate 2a with a yield of 83%; the synthetic reaction formula is as follows:

[0049]

[0050] 1 H NMR (400MHz, Chloroform-d): δ8.39 (d, J = 0.7Hz, 2H), 7.75-7.69 (m, 4H), 7.31 (d, J = 8.3Hz, 2H), 2.71 (s, 6H); such as Figure 3 As shown; 13C 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; Figure 4 HRMS (ESI) calculation for C 22 H 14 O6Se[M+Na] + 470.9907, found 470.9918.

[0051] Example 3

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

[0053] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromoacetophenone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3a; a brown-yellow solid with a yield of 73%; the structural formula of compound 3a is as follows:

[0054]

[0055] 1 H NMR (400MHz, DMSO-d6): δ11.40(s,2H),8.12(s,2H),8.03(d,J=2.1Hz,2H),7.86(d,J=7.7Hz,4H ),7.69(dd,J=8.5,2.1Hz,2H),7.39(dd,J=8.3,4.7Hz,6H),7.30(d,J=7.5Hz,4H),2.26(s,6H); such as Figure 5 As shown; 13C NMR (100 MHz, DMSO-d6): 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; Figure 6 HRMS (ESI) calculation for C 40 H 28 N6O4S2Se[M+H] + 795.0911, found 795.0967.

[0056] Example 4

[0057] The preparation method of 6,6'-selenidebis(3-(-1-(2-(4-(3-chlorophenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one (3b) comprises the following steps:

[0058] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(3-chlorophenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3b; a brown-yellow solid with a yield of 77%; the structural formula of compound 3b is as follows:

[0059]

[0060] 1 H NMR (400MHz, DMSO-d6): δ11.38(s,2H),8.07(s,2H),7.95(s,2H),7.88(s,2H), 7.78(d,J=7.8Hz,2H),7.64(d,J=8.6Hz,2H),7.44-7.28(m,8H),2.23(s,6H); such as Figure 7 As shown; 13C NMR (100 MHz, DMSO-d6)): 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; Figure 8 HRMS (ESI) calculation for C 40 H 26 Cl2N6O4S2Se[M+H] + 863.0132, found863.0222.

[0061] Example 5

[0062] The preparation method of 6,6'-selenidebis(3-(1-(2-(4-(3-methoxyphenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one) (3c) comprises the following steps:

[0063] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(3-methoxyphenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3c; a brown-yellow solid with a yield of 59%; the structural formula of compound 3c is as follows:

[0064]

[0065] 1 H NMR (400MHz, DMSO-d6): δ11.34(s,2H),8.07(s,2H),7.96(d,J=2.2Hz,2H),7.64(dd,J=8.6,2.1Hz,2H),7.42(s,2H) ,7.34(d,J=8.6Hz,2H),7.29(d,J=7.4Hz,4H),6.88-6.81(m,4H),3.80(d,J=1.9Hz,0H),3.77(s,6H),2.24(s,6H); such as Fig. 9 As shown; 13C NMR (100 MHz, DMSO-d6): 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; Fig.10 HRMS (ESI) calculation for C 42 H 32 N6O6S2Se[M+H] + 855.1122, found855.1091.

[0066] Example 6

[0067] The preparation method of 6,6'-selenidebis(3-(1-(2-(4-(4-methoxyphenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one) (3d) comprises the following steps:

[0068] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(4-methoxyphenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3d; a brown-yellow solid with a yield of 64%; the structural formula of compound 3d is as follows:

[0069]

[0070] 1 H NMR (400MHz, DMSO-d6): δ11.35(s,2H),8.10(s,2H),8.01(d,J=2.1Hz,2H),7.81-7.73(m,4H),7.67(dd ,J=8.6,2.1Hz,2H),7.38(d,J=8.6Hz,2H),7.11(s,2H),6.99-6.91(m,4H),3.77(s,6H),2.25(s,6H); such as Fig.11 As shown; 13C NMR (100 MHz, DMSO-d6): 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; Fig.12 HRMS (ESI) calculation for C 42 H 32 N6O6S2Se[M+H] + 855.1122, found 855.1164.

[0071] Example 7

[0072] The preparation method of 6,6'-selenidebis(3-(1-(2-(4-(2-hydroxyphenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one) (3e) comprises the following steps:

[0073] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(2-hydroxyphenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3e; a brown-yellow solid with a yield of 64%; the structural formula of compound 3e is as follows:

[0074]

[0075] 1 H NMR (400MHz, DMSO-d6): δ11.39(d,J=73.8Hz,2H),8.12(s,2H),8.00(d,J=2.1Hz,2H),7.79(dd,J=7.8,1.7 Hz,2H),7.67(dd,J=8.6,2.1Hz,2H),7.38(d,J=8.1Hz,4H),7.15(s,2H),6.90-6.81(m,6H),2.27(s,6H); such as Fig.13 As shown; 13C NMR (100 MHz, DMSO-d6): 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; Fig.14 HRMS (ESI) calculation for C 40 H 28 N6O6S2Se[M+H] + 827.0809, found 827.1199

[0076] Example 8

[0077] The preparation method of 6,6'-selenobis(3-(1-(2-(4-(3-hydroxyphenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one) (3f) comprises the following steps:

[0078] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(3-hydroxyphenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3f; a brown-yellow solid with a yield of 56%; the structural formula of compound 3f is as follows:

[0079]

[0080] 1 H NMR (400MHz, DMSO-d6): δ11.35(s,2H),9.47(s,2H),8.11(s,2H),8.01(d,J=2.1Hz,2H),7.68(dd,J=8.6,2.1 Hz, 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); such as Fig.15 As shown; 13C NMR (100 MHz, DMSO-d6): δ 169.52, 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; Fig.16 HRMS (ESI) calculation for C 40 H 28 N6O6S2Se[M+Na] + 849.0629, found 849.0694.

[0081] Example 9

[0082] The preparation method of 6,6'-selenidebis(3-(1-(2-(4-(4-methylphenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one) (3g) comprises the following steps:

[0083] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(4-methylphenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain 3 g of coumarin selenoether thiazole hydrazone derivatives; brown-yellow solid, yield 71%; the structural formula of compound 3g is as follows:

[0084]

[0085] 1 H NMR (400MHz, DMSO-d6): δ11.37(s,2H),8.12(s,2H),8.03(d,J=2.2Hz,2H),7.76- 7.67(m,6H),7.39(d,J=8.6Hz,2H),7.24-7.17(m,6H),2.31(s,6H),2.25(s,6H); such as Fig.17 As shown; 13C NMR (100 MHz, DMSO-d6): δ 169.60, 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; Fig.18 HRMS (ESI) calculation for C 42 H 32 N6O4S2Se[M+H] + 823.1224, found823.1266.

[0086] Example 10

[0087] The preparation method of 6,6'-selenobis(3-(1-(2-(4-(4-fluorophenyl)thiazol-2-yl)hydrazide)ethyl)-2H-benzopyran-2-one)(3h) comprises the following steps:

[0088] The intermediate 2a (0.1356 g, 0.3 mmol) of Example 2, thiosemicarbazide (0.0574 g, 0.63 mmol) and 5 mL of anhydrous ethanol were added to a 15 mL high pressure reaction tube equipped with a magnetic stirrer, and 2 drops of acetic acid were added dropwise. After heating on a heating plate at 80 ° C. for 24 h, 0.63 mmol of 2-bromo-1-(4-fluorophenyl)ethanone was directly added to the reaction tube. The reaction progress was monitored by TLC. The reaction was completed in about 12 h, cooled to room temperature, and the reaction solution was filtered off under reduced pressure. It was washed with deionized water and anhydrous ethanol for 3 times, respectively, and dried. The dried crude product was then crystallized with pyridine and anhydrous ethanol (volume ratio of 1:10) to obtain a coumarin selenoether thiazole hydrazone derivative 3h; a brown-yellow solid with a yield of 78%; the structural formula of compound 3h is as follows:

[0089]

[0090] 1 H NMR (400MHz, DMSO-d6): δ11.39(s,2H),8.11(s,2H),8.01(d,J=2.1Hz,2H),7.90-7.85(m,4 H),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); such as Fig.19 As shown; 13C NMR (100 MHz, DMSO-d6): δ 169.79, 163.27, 160.84, 159.28, 153.36, 145.22, 140.35, 136.84, 133.63, 127.95 (d, J = 8.1 Hz), 127.59, 126.16, 120.65, 117.88, 116.02, 115.80, 104.58, 16.63; Fig. 20 As shown; 19 F NMR (376MHz, DMSO-d6): δ-114.56; Fig.21 HRMS (ESI) calculation for C 40 H 26 F2N6O4S2Se[M+H] + 831.0723, found831.1199.

[0091] Embodiment 11

[0092] The coumarin selenoether thiazole hydrazone compounds prepared in Examples 3-10 were tested for antioxidant activity.

[0093] 1. Determination of DPPH free radical scavenging rate

[0094] Add 100 μL of DPPH ethanol solution (25 μg / mL) to a 96-well ELISA plate, and then add 100 μL of sample solutions of different concentrations (200, 100, 50, 25, 12.5, 6.25 μmol / L) to each well. React in the dark at room temperature for 30 min. Use an ELISA reader to measure the absorbance value A1 of the sample group at a wavelength of 517 nm. Set up 3 parallels for each group, and L-ascorbic acid is a positive control.

[0095] DPPH free radical scavenging rate = [(A3-A0)-(A1-A2)] / (A3-A0)*100%; the absorbance value A2 of the sample blank group was 100 μL sample solution + 100 μL ethanol solvent, the absorbance value A3 of the negative control group was 100 μL DPPH ethanol solution + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent, and the absorbance A0 of the blank group was 100 μL ethanol + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent.

[0096] 2. Determination of free radical scavenging rate of Galvinoxyl

[0097] Add 100 μL of Galvinoxyl ethanol solution (30 μg / mL) to a 96-well ELISA plate, and then add 100 μL of sample solutions of different concentrations (50, 40, 30, 20, 10, 5 μmol / L) to each well, respectively. React in the dark at room temperature for 30 min, and use an ELISA reader to measure the absorbance value A1 of the sample group at a wavelength of 428 nm. Set up 3 parallels for each group, and L-ascorbic acid is a positive control.

[0098] Galvinoxyl free radical scavenging rate = [(A3-A0)-(A1-A2)] / (A3-A0)*100%; the absorbance value A2 of the sample blank group was 100 μL sample solution + 100 μL ethanol, the absorbance value A3 of the negative control group was 100 μL Galvinoxyl ethanol solution + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent, and the absorbance A0 of the solvent blank group was 100 μL ethanol + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent.

[0099] 3. Determination of ABTS cation radical scavenging

[0100] Take 10mg ABTS and 3mg K2S2O8 and add them to a 10mL volumetric flask, add deionized water to dilute and fix the volume, place it in the dark at room temperature for 24h to prepare ABTS working solution, and dilute it with anhydrous ethanol after oxidation to make the absorbance of ABTS solution at 734nm (0.700±0.020). Add 100μLABTS working solution to a 96-well ELISA plate, and then add 100μL of sample solutions of different concentrations (80, 60, 40, 20, 10, 5μmol / L) to each well, react at room temperature in the dark for 12min, and use an ELISA reader to measure the absorbance value A1 of the sample group at a wavelength of 734nm. Set up 3 parallels for each group, and L-ascorbic acid is a positive control.

[0101] ABTS + Free radical scavenging rate = [(A3-A0)-(A1-A2)] / (A3-A0)*100%; the absorbance value A2 of the sample blank group is 100 μL sample solution + 100 μL ethanol, the absorbance value A3 of the negative control group is 100 μL ABTS ethanol solution + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent, and the absorbance A0 of the solvent blank group is 100 μL ethanol + 100 μL (DMSO:EtOH=1:5, volume ratio) mixed solvent.

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

[0103] Fig. 22 is the DPPH·scavenging rate of the coumarin selenoether thiazole hydrazone compounds prepared in Examples 3-10 of the present invention. Fig. 22 As shown in Figure 2, all eight compounds 3a to 3h have the ability to scavenge DPPH free radicals. Except for 3b and 3h, the IC 50 The values ​​were lower than the IC of L-ascorbic acid 50 The value was 12.56 μmol / L, indicating that its antioxidant effect was stronger than that of L-ascorbic acid. Among them, compound 3e had the strongest activity, and its IC 50 The value is 7.47 μmol / L; the IC 50 The results showed that the compounds substituted with methoxy and phenolic hydroxyl groups (3c, 3d, 3e, 3f) had stronger DPPH radical scavenging ability than the compounds substituted with methyl and halogen atoms (3b, 3g, 3h). It is speculated that the compounds containing electron-donating groups may have stronger DPPH radical scavenging ability than the compounds containing electron-withdrawing groups.

[0104] Fig.23 The galvinoxyl·scavenging rate of the coumarin selenoether thiazole hydrazone compounds prepared in Examples 3-10 of the present invention. Fig.23 As shown in Table 1, except for compound 3d, the scavenging rates of the other compounds on Galvinoxyl free radicals at 25 μmol / L were stronger than those of L-ascorbic acid. It can be seen from Table 1 that the IC 50 The value is 4.62~11.16μmol / L, which is similar to the IC of L-ascorbic acid. 50 The value of 8.56 μmol / L was similar to that of L-ascorbic acid, indicating that its antioxidant effect was comparable to that of L-ascorbic acid. Among them, compound 3e had the strongest activity, and its IC value for scavenging Galvinoxyl free radicals was 50 The value is 4.62μmol / L.

[0105] Fig.24 ABTS of coumarin selenoether thiazole hydrazone compounds prepared in Examples 3-10 of the present invention + · Clearance rate. Fig.24 As shown in Table 1, all eight compounds 3a to 3h have the ability to scavenge ABTS cation radicals, among which compounds 3e, 3f, and 3g have stronger scavenging rates for ABTS cation radicals than L-ascorbic acid at 40 μmol / L. As shown in Table 1, the IC 50The IC50 values ​​of compounds 3a, 3e, 3f, and 3g were all lower than the IC50 values ​​of L-ascorbic acid. 50 The value was 13.94 μmol / L, indicating that the antioxidant effects of compounds 3a, 3e, 3f, and 3g were stronger than those of L-ascorbic acid.

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

[0107]

[0108]

[0109] Embodiment 11

[0110] Myeloperoxidase (encoding 4C1M) was downloaded as the receptor protein through the protein database retrieval at https: / / www.rcsb.org / . The downloaded protein was pre-processed by deleting duplicate chains, deleting water molecules, adding hydrogen atoms and minimizing energy using MOE software. A small molecule database of coumarin selenoether thiazole hydrazone compounds was established. The compounds in the small molecule database were molecularly docked with the processed proteins through the Compute-Dock operation. The smaller the docking score, the less energy is required for docking, and the more spontaneous the reaction is. After docking, the Ligand Interactions operation can intuitively display information such as whether the docked compound has side chain exposure, the binding mode of the compound with the key amino acids in the active center, and the interaction force between the compound and the enzyme.

[0111] Compounds 3a~3h were docked with myeloperoxidase using MOE software to preliminarily explore their interactions and optimal docking conformations. Taking compound 3e as an example, Fig.25 FIG. 3 is a diagram showing the binding mode of compound 3e of the present invention to the active site of myeloperoxidase (MPO). Fig.25 As shown, the interaction between compound 3e and myeloperoxidase residues can be observed, and the docking score of the two is -7.6489 (Table 2). There are 5 amino acids involved in the interaction. Compound 3e forms hydrogen bonds with Ser266 and Asn186 through the phenolic hydroxyl group in the form of H-donor, forms pi-H interaction with Asn189 through the thiazole ring, and forms pi-H interaction with Asn192 and Gln193 through the benzene ring on the coumarin mother core. The docking score is a negative value, and the smaller the score, the tighter the compound binds to the protein. Table 2 lists the docking results of 3a to 3h, indicating that these 8 compounds have good affinity with myeloperoxidase in theory and have the potential to be myeloperoxidase inhibitors.

[0112] Table 2 Docking effect of compounds 3a-3h on myeloperoxidase

[0113]

[0114]

[0115] In summary, the present invention synthesized a total of 8 compounds of coumarin selenoether thiazole hydrazone derivatives, and investigated the in vitro antioxidant properties of coumarin selenoether thiazole hydrazone derivatives and their molecular mechanism of action for inhibiting myeloperoxidase. The in vitro antioxidant results showed that: in the three test systems, the 8 coumarin selenoether thiazole hydrazone derivatives all had good free radical scavenging ability, indicating that this type of compound is a potential antioxidant; molecular docking results showed that coumarin selenoether thiazole hydrazone derivatives mainly bind to myeloperoxidase through pi-H interaction and hydrogen bonding 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 the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the attached claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall 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 equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A coumarin selenoether thiazole hydrazone derivative, characterized in that: It has a chemical structure as shown in Formula I: Wherein, R is selected from substituted or unsubstituted phenyl, and the substituent of phenyl is selected from C1-C4 alkyl, alkoxy, hydroxyl or halogen.

2. The coumarin selenoether thiazole hydrazone derivative according to claim 1, characterized in that: The substituents of the phenyl group are selected from methyl, methoxy, hydroxy, chlorine or fluorine.

3. The coumarin selenoether 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 method for preparing the coumarin selenoether thiazole hydrazone derivative according to any one of claims 1 to 3, characterized in that: The following steps are involved: The intermediate 2a and thiosemicarbazide are dissolved in ethanol for addition condensation reaction. After the reaction, a bromoacetophenone compound is added to carry out a Hantzch cyclization reaction. After the reaction, vacuum filtration, washing, drying and crystallization are carried out in sequence to obtain a coumarin selenoether thiazole hydrazone compound. The synthetic reaction formula is shown below:

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

6. The method for preparing the coumarin selenoether thiazole hydrazone derivatives according to claim 4, characterized in that: The preparation method of the intermediate 2a comprises the following steps: The intermediate 1a and ethyl acetoacetate are dissolved in ethanol, and piperidine is added to carry out Knoevenagel condensation reaction. After the reaction is completed, vacuum filtration, washing, drying, and chromatography separation are carried out in sequence to obtain the intermediate 2a. The synthetic reaction formula is shown below:

7. The method for preparing the coumarin selenoether thiazole hydrazone derivatives 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, and the temperature of the Knoevenagel condensation reaction is 80° C. and the time is 1 h.

8. The method for preparing the coumarin selenoether thiazole hydrazone derivatives according to claim 6, characterized in that: The preparation method of the intermediate 1a comprises the following steps: Salicylaldehyde and selenium dioxide are dissolved in hydrochloric acid for selenization reaction. After the reaction, the intermediate 1a is obtained by filtering, washing, dissolving and recrystallizing in sequence. The synthetic reaction formula is shown below:

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

10. Use of the coumarin selenoether thiazole hydrazone derivative according to claim 1 in the preparation of antioxidant drugs, characterized in that: The antioxidant drug is used to inhibit myeloperoxidase.

Images