Compound for preventing and treating oxidative damage caused by ischemia-reperfusion and preparation method thereof

Abstract

The present invention provides a compound represented by formula A which can be used as a compound for preventing and treating oxidative damage caused by ischemia-reperfusion and a preparation method thereof. 10 As defined herein. The compounds of the present invention can react with peroxynitrite in the blood and further release carbon monoxide, thereby achieving anti-inflammatory and antioxidant effects. Tests of such compounds at the cellular level and in vivo level in mice show that they have achieved good therapeutic effects on a number of key evaluation indicators including brain near-death area, brain edema, blood-brain barrier, and neurological function scores, and have important potential for drug development.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry and relates to a compound for preventing and treating ischemia-reperfusion oxidative damage and a preparation method thereof. Background Art

[0002] Ischemic stroke accounts for 85% of all stroke cases. The medical treatment method is to use mechanical or drug methods to dissolve blood clots and promote blood reperfusion. However, ischemic stroke reperfusion is often accompanied by strong oxidative stress, leading to damage to vascular endothelial cells and distal organ failure. There are no drugs that can be used clinically to protect against oxidative damage caused by ischemia-reperfusion. Therefore, it is urgent to develop drugs that can be used to protect against oxidative damage caused by ischemia-reperfusion. Summary of the invention

[0003] The present invention provides a class of small molecule compounds, which can react with peroxynitrite in the blood and further release carbon monoxide, thereby achieving anti-inflammatory and antioxidant effects. Tests of the compounds at the cellular level and in vivo level of mice show that they have achieved good therapeutic effects on multiple key evaluation indicators including brain near-death area, brain edema, blood-brain barrier, and neurological function scores, and have important potential for drug development.

[0004] Specifically, the present invention provides a compound represented by formula A:

[0005]

[0006] In formula A, R1, R2, R3, R4, R5 and R6 are each independently selected from H, hydroxyl, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy and -NR 17 R 18 , where R 17 and R 18 R7, R8, R9 and R 10 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0007] In one or more embodiments, in Formula A, R1 and R2 are each independently selected from H, hydroxy, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, and -NR 17 R 18 , R3, R4, R5 and R6 are H.

[0008] In one or more embodiments, in Formula A, R7, R8, R9 and R 10 For H.

[0009] In one or more embodiments, the compound has a structure shown in Formula B:

[0010]

[0011] In formula B, R 11 and R 12 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0012] In one or more embodiments, in Formula B, R 11 and R 12 Each is independently selected from H, methyl and ethyl.

[0013] In one or more embodiments, the compound has a structure shown in Formula C:

[0014]

[0015] In formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0016] In one or more embodiments, in Formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H, methyl and ethyl.

[0017] In one or more embodiments, the compound is selected from:

[0018]

[0019] The present invention also provides a method for preparing a compound as described in any embodiment herein, the method comprising:

[0020] (1) reacting a compound of formula D with a compound of formula E to obtain a compound of formula F;

[0021]

[0022] Among them, R1-R 10 As described in any embodiment of the text;

[0023] (2) reacting the compound of formula F under the action of boron tribromide to obtain the compound of formula A.

[0024] In one or more embodiments, in step (1), the compound of formula D is first reacted with the compound of formula E in tetrahydrofuran, and then the product is reacted in toluene under the action of p-toluenesulfonic acid.

[0025] In one or more embodiments, in step (2), the reaction solvent is dichloromethane.

[0026] The present invention also provides the use of the compound described in any embodiment of the present invention in the preparation of a drug for preventing and treating ischemia-reperfusion oxidative damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The dose-effect relationship diagram of the reaction between PCOD585 (1M) and ONOO-, the solvent is neutral phosphate buffer (50mM, pH=7.4, containing 5% DMF). Figure 1 A in FIG. 1 is a UV-visible titration spectrum, and the ONOO- concentration range is 0-12M. Figure 1 B in the figure is the change of the absorption value at 560nm in the UV-visible titration spectrum with the ONOO- concentration. Figure 1 C in the figure is the fluorescence emission titration spectrum, the excitation light is 560nm, and the ONOO- concentration range is 0-12M. Figure 1 D in the figure is the change of fluorescence emission intensity at 585 nm in the fluorescence emission titration spectrum with ONOO- concentration.

[0028] Figure 2 This is the result of real-time monitoring of the release of carbon monoxide by the reaction of PCOD585 and ONOO- using GC-MS.

[0029] Figure 3 The reaction kinetics of PCOD585 (1 μM) and ONOO- is shown in the figure. The excitation light is 560 nm, the emission light is 585 nm, and one or ten equivalents of ONOO are added at one time. - , observe the change of fluorescence intensity in the solution over time.

[0030] Figure 4 This is a graph showing the reaction selectivity of PCOD585.

[0031] Figure 5 These are confocal images of EA.hy926 cells, where "a" is the control experimental group, "b" is the SIN-1 drug treatment group, "c" is the glucose-hypoxia cell group, "1" is the PCOD585 (1μM) channel (excitation 561nm, emission 570-620nm), "2" is the COP-1 (0.1μM) channel (excitation 488nm, emission 500-555nm), and "3" is the superposition of the two channels. The scale is 50μm.

[0032] Figure 6The results of TTC brain tissue staining analysis, where "Sham" is the control group, "MCAO" is the ischemia-reperfusion mouse model (ischemia for 2 hours, reperfusion for 22 hours), "-" is the untreated group, and "PCOD585" is the PCOD585-treated group (concentrations are 3, 6, and 9 mg / kg, respectively). * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001, and **** indicates p<0.0001. n=3.

[0033] Figure 7 The results of TTC brain tissue staining analysis are shown, where "Sham" is the control group, "MCAO" is the ischemia-reperfusion mouse model (ischemia for 2 hours, reperfusion for 22 hours), "-" is the untreated group; "PCOD585" is the PCOD585-treated group (concentration of 6 mg / kg), "3" represents the compound 3-treated group (concentration of 6 mg / kg), "9" represents the compound 9-treated group (concentration of 6 mg / kg), * represents p<0.05, ** represents p<0.01, *** represents p<0.001, **** represents p<0.0001, and n=3. DETAILED DESCRIPTION

[0034] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used in the text are the common meanings understood by those skilled in the art for the present invention. In the event of a conflict, the definition in this specification shall prevail.

[0035] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0036] In this article, all features such as values, quantities, contents and concentrations defined in the form of numerical ranges or percentage ranges are only for brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be considered to have included and specifically disclosed all possible secondary ranges and individual values ​​within the range (including integers and fractions).

[0037] Herein, when describing embodiments or examples, it should be understood that they are not used to limit the present invention to these embodiments or examples. On the contrary, all substitutes, improvements and equivalents of the methods and materials described in the present invention can be included in the scope limited by the claims.

[0038] In this document, in order to make the description concise, not all possible combinations of various technical features in various embodiments or examples are described. Therefore, as long as there is no contradiction in the combination of these technical features, the various technical features in various embodiments or examples can be combined arbitrarily, and all possible combinations should be considered to be within the scope of this specification.

[0039] The present invention provides a class of compounds having the function of preventing and treating oxidative damage caused by ischemia-reperfusion, which has a structure shown in the following formula A:

[0040]

[0041] In formula A, R1, R2, R3, R4, R5 and R6 are each independently selected from H, hydroxyl, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy and -NR 17 R 18 , where R 17 and R 18 R7, R8, R9 and R 10 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0042] As used herein, alkyl includes straight and branched chain alkyl groups which may contain 1 to 4 carbon atoms, such as 1, 2, 3 or 4 carbon atoms.

[0043] Herein, halogen includes fluorine, chlorine, bromine and iodine.

[0044] Herein, the halogen-substituted alkyl and alkoxy groups may be alkyl and alkoxy groups in which one or more or all hydrogen atoms, for example, 1 to 3 hydrogen atoms, are substituted with halogen.

[0045] In some embodiments, in Formula A, R1 and R2 are each independently selected from H, hydroxy, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, and -NR 17 R 18 In some embodiments, in Formula A, R1 and R2 are each independently selected from H, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy and -NR 17 R 18 .

[0046] In some embodiments, in Formula A, R3, R4, R5 and R6 are H.

[0047] In some embodiments, in Formula A, R7, R8, R9 and R 10Each is independently selected from H and C1-C4 alkyl. In some embodiments, in Formula A, R7, R8, R9 and R 10 For H.

[0048] In some embodiments, the structure of the compound of formula A of the present invention is as shown in formula B:

[0049]

[0050] In formula B, R 11 and R 12 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0051] In some embodiments, in Formula B, R 11 and R 12 Each is independently selected from H and C1-C4 alkyl. In some embodiments, in Formula B, R 11 and R 12 Each is independently selected from H, methyl and ethyl.

[0052] In some embodiments, the structure of the compound of formula A of the present invention is shown in formula C:

[0053]

[0054] In formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

[0055] In some embodiments, in Formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H and C1-C4 alkyl. In some embodiments, in Formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H, methyl and ethyl.

[0056] In some embodiments, the compound of formula A of the present invention is selected from:

[0057]

[0058] The compound of formula A of the present invention can be prepared by a method comprising the following steps:

[0059] (1) reacting a compound of formula D with a compound of formula E to obtain a compound of formula F;

[0060]

[0061] Among them, R1-R 10 As described in any embodiment of the present invention;

[0062] (2) reacting the compound of formula F under the action of boron tribromide to obtain the compound of formula A.

[0063] The present invention also includes various intermediates prepared in the process of preparing the compound of formula A of the present invention, including but not limited to compounds of formula D, formula F and formula F.

[0064] In step (1), the compound of formula D and the compound of formula E can be reacted in a solvent, such as tetrahydrofuran, and this reaction can be carried out at a low temperature (e.g., -70°C to -80°C); then the reaction product of the compound of formula D and the compound of formula E is reacted in a solvent, such as toluene, under the action of an acid, such as p-toluenesulfonic acid (PTSA), to obtain a compound of formula F, and this reaction can be carried out under heating conditions (e.g., 110±10°C). The compound of formula D can be prepared by (wherein, R1-R6 are as described in any embodiment herein, and X is a halogen, such as bromine) and a lithium reagent (such as butyl lithium).

[0065] In some embodiments, in step (2), the reaction solvent is dichloromethane. The reaction in step (2) can be carried out at low temperature to room temperature (eg, about 0° C.-25° C.).

[0066] Examples 1-3 of the present application show the preparation process of some compounds of formula A. Based on these processes, those skilled in the art can appropriately change the starting materials and corresponding reaction conditions according to the actual compounds to be prepared to prepare the compounds of formula A of the present invention.

[0067] The compounds of the present invention are suitable for use as compounds for preventing and treating oxidative damage caused by ischemia-reperfusion. Therefore, the present invention provides the use of the compounds of the present invention as a drug for preventing and treating oxidative damage caused by ischemia-reperfusion, or the use in the preparation of a drug for preventing and treating oxidative damage caused by ischemia-reperfusion. The compounds of the present invention can be used as a drug for preventing or treating various diseases that benefit from preventing, inhibiting or alleviating ischemic stroke reperfusion, including oxidative stress caused by ischemic stroke reperfusion, damage to vascular endothelial cells, and distal organ failure.

[0068] The compounds of the present invention can react with peroxynitrite (ONOO -) reacts and further releases carbon monoxide. Therefore, the present invention provides the use of the compounds of the present invention as peroxynitrite acceptors or carbon monoxide donors. In some embodiments, as peroxynitrite acceptors or carbon monoxide donors, the compounds of the present invention can be used to study the involvement of peroxynitrite or carbon monoxide in physiological and pathological processes.

[0069] Therefore, the present invention also provides a composition containing the compound of formula A of the present invention. The composition may also contain other ingredients, such as solvents, including organic solvents and / or inorganic solvents. In certain embodiments, the solvent is water, DMF, phosphate buffer or a neutral phosphate buffer containing DMF. When used as a drug, the composition can be a pharmaceutical composition, which can contain various pharmaceutically acceptable carriers or excipients, fillers, lubricants, sweeteners, preservatives, etc. known in the art. The amount of the compound of the present invention in the pharmaceutical composition should be sufficient to provide an effective therapeutic or preventive effect, which can be determined by those skilled in the art according to the specific disease to be treated, the patient's gender, age and weight, etc.

[0070] Therefore, the present invention also provides the use of the compound of formula A in the preparation of a drug for treating or preventing various diseases in which a subject benefits from preventing, inhibiting or alleviating ischemic stroke reperfusion. In certain embodiments, the disease that benefits from preventing, inhibiting or alleviating ischemic stroke reperfusion includes but is not limited to oxidative stress caused by ischemic stroke reperfusion, damage to vascular endothelial cells, and distal organ failure. In the present invention, the subject can be a mammalian subject, such as a human.

[0071] The present invention also provides a method for absorbing ONOO- or releasing CO in a biological matrix, the method comprising providing the compound of formula A of the present invention to the biological matrix. The biological matrix can be a living organism, or it can be an isolated biological tissue or biological cell. Therefore, the method can be an in vivo method or an in vitro method. When the in vivo method is implemented, for example, when it is implemented on an experimental animal (such as a mouse, rabbit, etc.) or a model animal, the method should meet general ethical requirements.

[0072] The present invention will be described below in the form of specific examples. It should be understood that these examples are merely illustrative and are not intended to limit the scope of the present invention. The methods, reagents and materials used in the examples are, unless otherwise stated, conventional methods, reagents and materials in the art. The raw material compounds in the examples can all be purchased through commercial routes.

[0073] Example 1: Compound 3 Preparation

[0074]

[0075] (1) Diphenyl ether (compound 1, 1.38 g, 1.5 equivalents, 8 mmol) was dissolved in 50 mL of anhydrous tetrahydrofuran and cooled to -78°C. 3.23 mL of n-butyl lithium in n-hexane solution (2.5 M, 1.5 equivalents, 8 mmol) was added to the reaction solution. After 30 minutes of reaction, a solution of compound 5 (1.10 g, 1 equivalent, 5.39 mmol) in anhydrous tetrahydrofuran was added. The reaction was continued for 2 hours. 10 mL of saturated ammonium chloride solution was added to the solution to quench the excess lithium reagent. The reaction solution was extracted three times with water and dichloromethane. The organic phase was collected and dried over anhydrous sodium sulfate. The inorganic salt was filtered and the filtrate was dried under reduced pressure to obtain a yellow oil. The yellow oil was dissolved in anhydrous toluene, p-toluenesulfonic acid (0.93 g, 1.0 equivalent, 5.39 mmol) was added, the reaction solution was heated to 110°C for 10 hours and then cooled, 5 ml of saturated NaHCO3 aqueous solution was added, the reaction solution was extracted with water and dichloromethane three times, the organic phase was collected, dried with anhydrous sodium sulfate, the inorganic salt was filtered and the filtrate was dried under reduced pressure. The crude product was separated by silica gel column chromatography (solvent system: petroleum ether: ethyl acetate = 15:1, V / V) to obtain compound 2 (1.20 g, orange solid, 62%).

[0076] 1 H NMR (600MHz, CDCl3) δ7.97(d,J=7.7Hz,1H),7.76(td,J=7.6,1.2Hz,1H),7.63-7.57(m,1H),7.34(d,J=7.8Hz,1H),7.31-7.27(m,2H),7.19 (dd,J=8.2,1.1Hz,2H),6.98(td,J=7.9,1.2Hz,2H),6.58(dd,J=7.8,1.5Hz,2H),4.01(td,J=6.1,3.9Hz,2H),3.51(td,J=6.1,4.0Hz,2H). 13 CNMR(151MHz, CDCl3)δ201.4,153.6,152.2,137.2,135.2,129.9,129.3,128.7,127.7,123.4,122.9,122.7,116.2,109.8,66.4.ESI-MS,m / z,[M+Na] + ,calcd.for C 23 H 16 O4Na + ,379.0946; found 379.0945.

[0077] (2) Compound 2 (500 mg, 1 equivalent, 1.40 mmol) was dissolved in 25 mL of anhydrous dichloromethane, and BBr3 (1.05 mL, 10.0 equivalent, 14.03 mmol) was slowly added at room temperature, and the reaction was continued for 15 minutes until the raw material was consumed. Water was slowly added to quench the reaction, and the reaction solution was extracted with water and dichloromethane three times. The organic phase was collected and dried over anhydrous sodium sulfate, the inorganic salt was filtered and the filtrate was dried under reduced pressure. The crude product was separated by silica gel column chromatography (solvent system: petroleum ether: ethyl acetate = 15:1, V / V) to obtain compound 3 (198 mg, red solid, 45%).

[0078] 1 H NMR(600MHz, CDCl3)δ8.08(d,J=7.8Hz,1H),7.89(td,J=7.7,1.2Hz,1H),7.71-7.65(m,1H),7.42(d, J=7.8Hz,1H),7.33-7.28(m,2H),7.28-7.25(m,3H),6.97-6.92(m,2H),6.46(dd,J=7.8,1.4Hz,2H). 13 CNMR(151MHz, CDCl3)δ196.4,190.2,152.2,151.1,139.7,137.8,123.0,129.8,128.6,128.2,123.8,123.8,121.8,117.5.ESI-MS,m / z,[M+Na] + ,calcd.for C 21 H 12 O3Na + ,335.0684; found 335.0686.

[0079] Example 2: PCOD585 Preparation

[0080]

[0081] (1) Compound 5 (1.23 g, 1.0 equivalent, 6.0 mmol) was dissolved in anhydrous tetrahydrofuran and cooled to -78°C. Diphenyl ether monolithium reagent (Compound 4, 1 M, 1.5 equivalent, 7.2 mmol) was slowly added to the above solution and the reaction was continued for 2 hours. After compound 5 was completely consumed, 10 ml of saturated ammonium chloride solution was added to the solution to quench the excess lithium reagent. The reaction solution was extracted three times with water and dichloromethane. The organic phase was collected and dried over anhydrous sodium sulfate. The inorganic salt was filtered and the filtrate was decompressed and dried to obtain a brown oil. The brown oil was dissolved in anhydrous toluene, p-toluenesulfonic acid (0.37 g, 1.1 equivalent, 2.13 mmol) was added, the reaction solution was heated to 110°C for 10 hours, cooled, and 5 ml of saturated NaHCO3 aqueous solution was added. The reaction solution was extracted three times with water and dichloromethane. The organic phase was collected and dried over anhydrous sodium sulfate. The inorganic salt was filtered and the filtrate was decompressed and dried to dryness. The crude product was separated by silica gel column chromatography (solvent system: petroleum ether:ethyl acetate=10:1, V / V) to obtain compound 6 (0.83 g, orange solid, 86%).

[0082] 1 H NMR (600MHz, CDCl3) δ7.90(d,J=7.7Hz,1H),7.68(dd,J=11.0,3.9Hz,1H),7.51(t,J=7.4Hz,1H),7.34(d,J=7.8Hz,1H),6.50-6 .34(m,4H),6.27(s,2H),4.26-3.97(m,2H),3.56(td,J=6.3,4.3Hz,2H),3.33(td,J=10.9,7.0Hz,8H),1.16(t,J=7.0Hz,12H). 13 C NMR (151MHz, CDCl3) δ202.3,154.8,153.4,148.1,136.7,135.1,130.3,128.5,12 7.4,122.8,110.1,109.8,106.7,98.1,66.2,51.4,44.3,12.6.ESI-MS,m / z,[M+H] + ,calcd.for C 31 H 35 N2O4 + ,499.2591; found 499.2598.

[0083] (2) Compound 6 (100 mg, 1 equivalent, 0.2 mmol) was dissolved in 50 ml of anhydrous dichloromethane, and boron tribromide (402 mg, 8.0 equivalent, 1.6 mmol) was added to the solution at room temperature. Stirring was continued for 15 minutes until the raw material was consumed. Water was slowly added to quench the reaction. The reaction solution was extracted with water and dichloromethane three times. The organic phase was collected and dried over anhydrous sodium sulfate. The inorganic salt was filtered and the filtrate was dried under reduced pressure. The crude product was separated by silica gel column chromatography (solvent system: petroleum ether: ethyl acetate = 10:1, V / V) to finally obtain compound PCOD585 (25 mg, brown solid, 28%).

[0084] 1 H NMR (600MHz, CDCl3) δ8.01(d,J=7.7Hz,1H),7.81(t,J=7.3Hz,1H),7.59(t,J=7.2Hz,1H),7. 43(d,J=7.8Hz,1H),6.46(s,2H),6.22(s,4H),3.32(q,J=6.9Hz,8H),1.15(t,J=6.9Hz,12H). 13 C NMR (151MHz, CDCl3) δ199.0,191.9,153.5,152.5,148.9,139.2,137.8,129.2, 129.0,128.6,123.1,109.7,107.4,99.0,50.3,44.5,12.7.ESI-MS,m / z,[M+H] + ,calcd.for C 29 H 31 N2O3 + ,455.2329; found 455.2336.

[0085] Example 3: Compound 9 Preparation

[0086]

[0087] (1) Compound 7 (1.00 g, 1.0 equivalent, 3.23 mmol) was dissolved in 50 mL of anhydrous tetrahydrofuran and cooled to -78°C. 1.94 mL of n-butyl lithium in n-hexane solution (2.5 M, 1.5 equivalent, 4.85 mmol) was added to the reaction solution. After 30 minutes of reaction, a solution of compound 5 (0.31 g, 1.5 equivalent, 4.85 mmol) in anhydrous tetrahydrofuran was added. The reaction was continued for 2 hours. 10 mL of saturated ammonium chloride solution was added to the solution to quench the excess lithium reagent. The reaction solution was extracted three times with water and dichloromethane. The organic phase was collected and dried over anhydrous sodium sulfate. The inorganic salt was filtered and the filtrate was dried under reduced pressure to obtain a yellow oil. The yellow oil was dissolved in anhydrous toluene, p-toluenesulfonic acid (0.56 g, 1.0 equivalent, 4.85 mmol) was added, the reaction solution was heated to 110°C for 10 hours and then cooled, 5 ml of saturated NaHCO3 aqueous solution was added, the reaction solution was extracted with water and dichloromethane three times, the organic phase was collected, dried with anhydrous sodium sulfate, the inorganic salt was filtered and the filtrate was dried under reduced pressure. The crude product was separated by silica gel column chromatography, the solvent system (petroleum ether: ethyl acetate = 15:1, V / V) to obtain compound 8 (500 mg, orange solid, 43%).

[0088] 1 H NMR (400MHz, CDCl3) δ7.93(d,J=7.6Hz,1H),7.74(t,J=7.0Hz,1H),7.57(t,J=7.5Hz,1H),7.30(d,J=7.8Hz,1H),6.70( d,J=2.5Hz,2H),6.54(dd,J=8.7,2.5Hz,2H),6.46(d,J=8.7Hz,2H),4.05-3.98(m,2H),3.81(s,6H),3.57-3.50(m,2H). 13 C NMR (151MHz, CDCl3) δ201.8,159.9,154.0,153.0,137.2,135.2,130.7,129.2,1 27.5,123.3,115.0,109.8,109.7,101.1,66.5,55.5,52.0.ESI-MS,m / z,[M+Na] + ,calcd.for C 25 H 20 O6Na + ,439.1158;found439.1155.

[0089] (2) Compound 8 (200 mg, 1 equivalent, 0.48 mmol) was dissolved in 25 mL of anhydrous dichloromethane, and BBr3 (0.46 mL, 10.0 equivalent, 4.80 mmol) was slowly added at room temperature, and the reaction was continued for 15 minutes until the raw material was consumed. Water was slowly added to quench the reaction, and the reaction solution was extracted with water and dichloromethane three times. The organic phase was collected and dried over anhydrous sodium sulfate, the inorganic salt was filtered and the filtrate was dried under reduced pressure. The crude product was separated by silica gel column chromatography (solvent system: petroleum ether: ethyl acetate = 15:1, V / V) to obtain compound 9 (78 mg, red solid, 44%).

[0090] 1 H NMR (600MHz, CDCl3) δ7.99 (d, J = 7.7Hz, 1H), 7.85 (t, J = 7.3Hz, 1H), 7.62 (t, J = 7.2Hz, 1H), 7. 55(d,J=7.8Hz,1H),7.53(d,J=7.6Hz,2H),6.61(s,2H),6.58(d,J=7.6Hz,2H),3.84(s,6H). 13 C NMR (151MHz, CDCl3) δ196.5,187.1,157.8,157.6 152.0,148.4,144.2,135.0,128.9,127.9,126.7,124.1,107.9,104.1,63.5,55.8.ESI-MS,m / z,[M+Na] + ,calcd.for C 23 H 16 O5Na + ,395.0895; found 395.0891.

[0091] Example 4: PCOD585 and ONOO - UV-Vis and Fluorescence Titration of the Reaction

[0092] PCOD585 was reacted with different concentrations (0-12 μM) of ONOO in neutral phosphate buffer (50 mM, pH = 7.4, containing 5% DMF). - The UV-visible absorption titration and fluorescence emission titration experiments of the reaction were performed, and the results were as follows Figure 1 As shown. It can be seen that PCOD585 and ONOO - After the reaction, there is significant absorption at 560nm, and the absorption value increases with the concentration of ONOO - The fluorescence intensity increases with the increase of ONOO concentration, and there is a significant fluorescence emission at 585nm. - Increases with increasing concentration.

[0093] Example 5: GC-MS detection of PCOD585 and ONOO - The reaction releases carbon monoxide

[0094] The PCOD585 was monitored in real time by gas chromatography-mass spectrometry (GC-MS) after the addition of ONOO - The relationship between the amount of carbon monoxide released and time is as follows: Figure 2 As shown, it was demonstrated that PCOD585 and ONOO - The reaction releases carbon monoxide.

[0095] Example 6: PCOD585 and ONOO - Reaction kinetics experiments

[0096] Add one or ten equivalents of ONOO to 1 μM PCOD585 at once. - , observe the change of fluorescence intensity at 585nm in the solution with time under 560nm excitation light, the results are as follows Figure 3 shown.

[0097] Example 7: Reaction selectivity experiment of PCOD585

[0098] The fluorescence intensity at 585 nm was measured after adding different substances to PCOD585. Figure 4 shown.

[0099] Example 8: Confocal imaging of EA.hy926 cells

[0100] Confocal imaging of EA.hy926 cells using PCOD585 was performed. Figure 5 shown.

[0101] Example 9: TTC brain tissue staining analysis

[0102] Different doses of PCOD585 were used for TTC brain tissue staining analysis. The results are shown in Figure 6 shown.

[0103] Example 10: TTC brain tissue staining analysis

[0104] PCOD585, compound 3 and compound 9 were used for TTC brain tissue staining analysis. The results are shown in Figure 7 shown.

Claims

1. The compound represented by formula A: In formula A, R1 and R2 are each independently selected from H, hydroxy, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy and -NR 17 R 18 , where R 17 and R 18 Each is independently selected from H and C1-C4 alkyl; R3, R4, R5, R6, R7, R8, R9 and R 10 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

2. The compound according to claim 1, characterized in that In formula A, R1 and R2 are each independently selected from H, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy and -NR 17 R 18 ; and / or In formula A, R3, R4, R5, R6, R7, R8, R9 and R 10 For H.

3. The compound according to claim 1, characterized in that The compound has a structure shown in formula B: In formula B, R 11 and R 12 Each is independently selected from H, C1-C4 alkyl and halogen-substituted C1-C4 alkyl.

4. The compound according to claim 3, characterized in that In formula B, R 11 and R 12 Each is independently selected from H, methyl and ethyl.

5. The compound according to claim 1, characterized in that The compound has a structure shown in Formula C: In formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H and C1-C4 alkyl.

6. The compound according to claim 5, characterized in that In formula C, R 13 , R 14 , R 15 and R 16 Each is independently selected from H, methyl and ethyl.

7. The compound according to claim 1, characterized in that The compound is selected from:

8. A method for preparing a compound according to any one of claims 1 to 7, characterized in that The method comprises: (1) reacting a compound of formula D with a compound of formula E to obtain a compound of formula F; Among them, R1-R 10 As claimed in claim 1; (2) reacting the compound of formula F under the action of boron tribromide to obtain the compound of formula A.

9. The method according to claim 8, characterized in that In step (1), the compound of formula D and the compound of formula E are first reacted in tetrahydrofuran, and then the product is reacted in toluene under the action of p-toluenesulfonic acid; and / or In step (2), the reaction solvent is dichloromethane.

10. Use of the compound according to any one of claims 1 to 7 in the preparation of a drug for preventing and treating oxidative damage caused by ischemia-reperfusion.

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