A compound capable of releasing HNO and preparation method thereof

Abstract

The present invention provides a compound that can hydrolyze and release HNO under physiological conditions. This compound can release HNO, an endogenous signaling molecule, providing a new donor compound for HNO research. Furthermore, the compound can simultaneously release a fluorescent molecular probe while releasing HNO, allowing fluorescence detection technology to monitor changes in fluorescence intensity during HNO release.

Description

Technical Field

[0001] The present invention relates to a compound with a new structure and a preparation method thereof, in particular to a compound capable of releasing HNO by hydrolysis and a preparation method thereof. Background Art

[0002] Hydrogen nitrosyl (HNO) is a sibling product of the redox reaction of nitric oxide (NO) and the simplest nitroxyl compound. In recent years, HNO has been extensively studied by numerous researchers both domestically and internationally for its unique therapeutic pharmacological advantages, along with its biological properties, reactivity, and pharmacological activities.

[0003] Over the past decade, the number of patients hospitalized for heart failure has continued to rise, both domestically and internationally. Over time, unhealthy lifestyles have increased the risk of heart failure, and the number of deaths from heart failure has also risen. The advent of various new medications has provided patients with both relief and improved symptoms. However, as life expectancy increases, the associated costs have also increased exponentially. Despite the emergence of various new medications, their inherent limitations and associated side effects mean that not every patient can fully adapt to them. Some heart failure patients often require treatment with vasodilators.

[0004] As an endogenous signaling molecule, HNO can effectively protect the cardiovascular system and improve ventricular diastolic and systolic function, thereby reducing cardiac workload and increasing cardiac output, thus playing a therapeutic role. HNO is currently hailed as a rising star that can completely cure heart failure and cardiovascular disease. However, there are few known compounds that can provide hydronitrosyl. As research on HNO continues to grow, currently known compounds are no longer sufficient to meet the research needs. Summary of the Invention

[0005] The purpose of the present invention is to address the deficiencies of the above-mentioned prior art and provide a new compound that can effectively release HNO and a preparation method thereof.

[0006] To achieve the above object, the technical solution adopted by the present invention is: a compound capable of releasing HNO, characterized in that the structural formula of the compound is as follows:

[0007]

[0008] Wherein, R is selected from the following groups:

[0009]

[0010] As a preferred embodiment of the compound capable of releasing HNO of the present invention, the compound can release hydronitrosyl at a pH of 7.0-12.

[0011] As a preferred embodiment of the compound capable of releasing HNO of the present invention, the compound can also release a fluorescent molecular probe.

[0012] As a preferred embodiment of the compound capable of releasing HNO of the present invention, the molar ratio of the hydronitrosyl released by the compound to the fluorescent molecular probe is 1:1.

[0013] In addition, another object of the present invention is to provide a method for synthesizing the above-mentioned compound capable of releasing HNO using 4-dimethylaminophenylazobenzenesulfonyl chloride as a raw material. To achieve this object, the technical solution adopted by the present invention is as follows:

[0014] When the R group in the structural formula of the compound capable of releasing HNO is H, the preparation method of the compound capable of releasing HNO comprises the following steps:

[0015] (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution.

[0016] (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution;

[0017] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained initial product was dried in a vacuum drying oven at room temperature for 24 to 48 hours to obtain the compound capable of releasing HNO.

[0018] When, in the structural formula of the compound capable of releasing HNO, R is a group other than H, the preparation method of the compound capable of releasing HNO comprises the following steps:

[0019] (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution.

[0020] (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution;

[0021] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained primary product was dried in a vacuum drying oven at room temperature for 24 to 48 hours to obtain an intermediate;

[0022] (4) Under N2 atmosphere and ice-water bath conditions, weigh the intermediate obtained in step (3) and pour it into a reaction flask, and add anhydrous THF to dissolve it;

[0023] (5) Add triethylamine to the reaction flask, then slowly add the acylating agent to the reaction flask. After 10 to 40 minutes, remove the ice-water bath and stir the reaction at room temperature for 5 to 10 hours.

[0024] (6) The solvent was dried using a rotary evaporator to obtain red solid particles, and then the crude product was dissolved in EtOAc, poured into a separatory funnel, and washed with water to remove water-soluble impurities. The obtained organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was again dried using a rotary evaporator to dry the solvent. The obtained preliminary product was dried in a vacuum drying oven at room temperature to obtain the compound capable of releasing HNO;

[0025] The acylating agent is selected from acetyl chloride, 2,2,2-trifluoroacetyl chloride, propionyl chloride, 2-pyridinecarbonyl chloride, pivaloyl chloride, 4-chlorobenzoyl chloride, 4-fluorobenzoyl chloride, 4-methoxybenzoyl chloride, 4-(tert-butyl)benzoyl chloride, 4-cyanobenzoyl chloride, 2-naphthoyl chloride, 3-chlorobenzoyl chloride, 3-fluorobenzoyl chloride, quinoline-2-carbonyl chloride, 3-methoxybenzoyl chloride, 2,4,6-trimethylbenzoyl chloride, At least one of benzoyl chloride, 4-nitrobenzoyl chloride, 3,5-difluorobenzoyl chloride, 4-bromo-3-methylbenzoyl chloride, benzoyl chloride, 4-(trifluoromethyl)benzoyl chloride, 2,6-difluorobenzoyl chloride, methyl 4-chloroformylbenzoate, 3-(trifluoromethoxy)benzoyl chloride, diphenylcarbamoyl chloride, 2-(4-chlorophenyl)-3-methylbutanoyl chloride, and 2-phenoxybenzoyl chloride.

[0026] As a preferred embodiment of the method for preparing the HNO-releasing compound of the present invention, the molar ratio of NH2OH·HCl to 4-dimethylaminophenylazobenzenesulfonyl chloride in step (1) is: NH2OH·HCl:4-dimethylaminophenylazobenzenesulfonyl chloride = 2.5-6.0:1. As a more preferred embodiment of the method for preparing the HNO-releasing compound of the present invention, the molar ratio of NH2OH·HCl to 4-dimethylaminophenylazobenzenesulfonyl chloride in step (1) is: NH2OH·HCl:4-dimethylaminophenylazobenzenesulfonyl chloride = 15.576:3.095.

[0027] In a preferred embodiment of the method for preparing a compound capable of releasing HNO according to the present invention, the molar ratio of NH2OH·HCl to triethylamine in step (1) is: NH2OH·HCl:triethylamine = 3.2-5:1 mmol / ml. In a more preferred embodiment of the method for preparing a compound capable of releasing HNO according to the present invention, the molar ratio of NH2OH·HCl to triethylamine in step (1) is: NH2OH·HCl:triethylamine = 15.576:3.406.

[0028] In a preferred embodiment of the method for preparing a compound capable of releasing HNO according to the present invention, the molar volume ratio of NH2OH·HCl to tetrahydrofuran in step (1) is: NH2OH·HCl:tetrahydrofuran = 1.01-1.6:1 mmol / ml. In a more preferred embodiment of the method for preparing a compound capable of releasing HNO according to the present invention, the molar volume ratio of NH2OH·HCl to tetrahydrofuran in step (1) is: NH2OH·HCl:tetrahydrofuran = 15.576:10 mmol / ml.

[0029] As a preferred embodiment of the method for preparing the compound capable of releasing HNO according to the present invention, step (1) is as follows: NH2OH·HCl (m=1.075 g, N=15.576 mmol, 5 eq.) and 473.2 μL of triethylamine (N=3.406 mmol 1.1 eq) are weighed and added into a reaction flask, stirred evenly, and then 10 ml of tetrahydrofuran is added. The stirring is continued until white turbidity appears in the reaction flask, and then 4-dimethylaminophenylazobenzenesulfonyl chloride (m=1.0 g, N=3.095 mmol) is weighed and slowly added into the reaction flask. The solution in the reaction flask turns red, and the reaction is carried out at room temperature for 16 h to obtain a red reaction solution.

[0030] As a preferred embodiment of the method for preparing the compound capable of releasing HNO of the present invention, the reaction in step (1) is carried out at room temperature for 14 to 18 hours. As a more preferred embodiment of the method for preparing the compound capable of releasing HNO of the present invention, the reaction in step (1) is carried out at room temperature for 16 hours.

[0031] As a preferred embodiment of the method for preparing the compound capable of releasing HNO of the present invention, in step (2), water washing and extraction can be performed in a separatory funnel. 50 ml of water can be used for each washing, and the washing can be performed 7 times.

[0032] As a preferred embodiment of the method for preparing the HNO-releasing compound of the present invention, the molar ratio of the acylating agent to the intermediate in step (5) is: acylating agent: intermediate = 1.1-1.3:1. As a more preferred embodiment of the method for preparing the HNO-releasing compound of the present invention, the molar ratio of the acylating agent to the intermediate in step (5) is: acylating agent: intermediate = 0.343:0.312.

[0033] As a preferred embodiment of the method for preparing the compound capable of releasing HNO of the present invention, the molar ratio of triethylamine to the intermediate in step (5) is: triethylamine:intermediate=1.1-1.3:1. As a more preferred embodiment of the method for preparing the compound capable of releasing HNO of the present invention, the molar ratio of triethylamine to the intermediate in step (5) is: triethylamine:intermediate=0.343:0.312.

[0034] Finally, the present invention also provides the use of the aforementioned HNO-releasing compounds in the preparation of medicaments for treating heart failure and cardiovascular diseases. The aforementioned compounds of the present invention are capable of effectively releasing HNO and can serve as HNO donor compounds, providing new HNO donors for HNO research. HNO, as an endogenous signaling molecule, can effectively protect the cardiovascular system, improve ventricular diastolic and systolic function, reduce cardiac workload, and increase cardiac output, thereby achieving a therapeutic effect. Furthermore, HNO can effectively inhibit the growth and reproduction of tumor cells. Therefore, the HNO donor compounds described in this application provide new pharmaceutical options for treating heart failure, cardiovascular diseases, and tumors.

[0035] The HNO-releasing compound provided by the present invention can effectively release HNO and, as an HNO donor compound, provides a new HNO donor for HNO research. The preparation method of the HNO-releasing compound of the present invention uses 4-dimethylaminophenylazobenzenesulfonyl chloride as a raw material, has simple process steps, and is easily scalable. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The synthetic route of one embodiment of the compound capable of releasing HNO according to the present invention is as follows;

[0037] Figure 2 A synthetic route for another embodiment of the compound capable of releasing HNO according to the present invention;

[0038] Figure 3 This is a hydrogen nuclear magnetic resonance spectrum of an embodiment of the compound capable of releasing HNO according to the present invention;

[0039] Figure 4 This is a carbon NMR spectrum of an embodiment of the compound capable of releasing HNO according to the present invention;

[0040] Figure 5 This is a mass spectrum of an embodiment of the compound capable of releasing HNO according to the present invention;

[0041] Figure 6 This is a kinetic degradation test diagram of an embodiment of the compound capable of releasing HNO according to the present invention;

[0042] Figure 7 This is a diagram of an HNO generation detection test of an embodiment of the compound capable of releasing HNO according to the present invention;

[0043] Figure 8 This is a fluorescence detection diagram of an embodiment of the compound capable of releasing HNO according to the present invention. DETAILED DESCRIPTION

[0044] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present application will be further described below in conjunction with specific embodiments.

[0045] The present invention discloses a compound capable of releasing HNO, and the structural formula of the compound is shown below:

[0046]

[0047] Wherein R is selected from the following groups

[0048]

[0049] When the above R selects H group, the synthetic route of the compound is as shown in the attached Figure 1 As shown, the preparation method comprises the following steps:

[0050] (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution.

[0051] (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution;

[0052] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained initial product was dried in a vacuum drying oven at room temperature for 24 to 48 hours to obtain the compound capable of releasing HNO.

[0053] When the above-mentioned R is selected from groups other than H, the synthetic route of the compound is as shown in the attached Figure 2 As shown, the preparation method comprises the following steps:

[0054] (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution.

[0055] (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution;

[0056] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained primary product was dried in a vacuum drying oven at room temperature for 24 to 48 hours to obtain an intermediate;

[0057] (4) Under N2 atmosphere and ice-water bath conditions, weigh the intermediate obtained in step (3) and pour it into a reaction flask, and add anhydrous THF to dissolve it;

[0058] (5) Add triethylamine to the reaction flask, then slowly add the acylating agent to the reaction flask. After 10 to 40 minutes, remove the ice-water bath and stir the reaction at room temperature for 5 to 10 hours.

[0059] (6) The solvent was dried using a rotary evaporator to obtain red solid particles, and then the crude product was dissolved in EtOAc, poured into a separatory funnel, and washed with water to remove water-soluble impurities. The obtained organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was again dried using a rotary evaporator to dry the solvent. The obtained preliminary product was dried in a vacuum drying oven at room temperature to obtain the compound capable of releasing HNO;

[0060] The acylating agent is selected from acetyl chloride, 2,2,2-trifluoroacetyl chloride, propionyl chloride, 2-pyridinecarbonyl chloride, pivaloyl chloride, 4-chlorobenzoyl chloride, 4-fluorobenzoyl chloride, 4-methoxybenzoyl chloride, 4-(tert-butyl)benzoyl chloride, 4-cyanobenzoyl chloride, 2-naphthoyl chloride, 3-chlorobenzoyl chloride, 3-fluorobenzoyl chloride, quinoline-2-carbonyl chloride, 3-methoxybenzoyl chloride, 2,4,6 -trimethylbenzoyl chloride, 4-nitrobenzoyl chloride, 3,5-difluorobenzoyl chloride, 4-bromo-3-methylbenzoyl chloride, benzoyl chloride, 4-(trifluoromethyl)benzoyl chloride, 2,6-difluorobenzoyl chloride, methyl 4-chlorobenzoylbenzoate, 3-(trifluoromethoxy)benzoyl chloride, diphenylcarbamoyl chloride, 2-(4-chlorophenyl)-3-methylbutanoyl chloride, 2-phenoxybenzoyl chloride.

[0061] When the above R is an H group, the compound (C2) capable of releasing HNO according to the present invention can be synthesized by a one-step method. When the above R is a group other than an H group, the compound (C2) is further reacted with different acylating agents as an intermediate to finally obtain the compound in which the above R is a group other than H.

[0062] Example 1

[0063] An embodiment of the compound capable of releasing HNO of the present invention, the structure of the compound capable of releasing HNO of this embodiment is shown below:

[0064]

[0065] The synthetic route of the compound capable of releasing HNO described in this embodiment is shown in the attached figure. Figure 1 As shown, the preparation method comprises the following steps:

[0066] (1) Weigh NH2OH·HCl (m=1.075 g, N=15.576 mmol, 5 eq.) and 473.2 μL triethylamine (N=3.406 mmol 1.1 eq) into a reaction flask, stir evenly, then add 10 ml tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride (m=1.0 g, N=3.095 mmol) and slowly add it into the reaction flask. The solution in the reaction flask turns red. React at room temperature for 16 h to obtain a red reaction solution.

[0067] (2) 80 ml of EtOAc was added to the reaction flask of step (1), and the mixture was washed with water (7 x 50 ml), and the resulting organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by filtration to obtain a light red reaction solution;

[0068] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained preliminary product was dried in a vacuum drying oven at room temperature for 24 h to obtain 0.977 g of the product, namely, the compound capable of releasing HNO.

[0069] Example 2

[0070] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0071]

[0072] Wherein R is the following group:

[0073]

[0074] The preparation method of the compound capable of releasing HNO described in this embodiment comprises the following steps:

[0075] (1) Weigh NH2OH·HCl (m=1.075 g, N=15.576 mmol, 5 eq.) and 473.2 μL triethylamine (N=3.406 mmol 1.1 eq) into a reaction flask, stir evenly, then add 10 ml tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride (m=1.0 g, N=3.095 mmol) and slowly add it into the reaction flask. The solution in the reaction flask turns red. React at room temperature for 16 h to obtain a red reaction solution.

[0076] (2) 80 ml of EtOAc was added to the reaction flask of step (1), and the mixture was washed with water (7 x 50 ml), and the resulting organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by filtration to obtain a light red reaction solution;

[0077] (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the resulting primary product was dried in a vacuum drying oven at room temperature for 24 h to obtain 0.977 g of the intermediate;

[0078] (4) Under N2 atmosphere and ice-water bath conditions, weigh the intermediate obtained in step (3) (100.0 mg, 0.312 mmol) and pour it into a reaction flask, and add anhydrous THF to dissolve;

[0079] (5) Add triethylamine (0.048 ml, 0.343 mmol) to the reaction flask, then slowly add acetyl chloride (0.343 mmol) to the reaction flask. After 10 to 40 minutes, remove the ice-water bath and stir the reaction at room temperature for 5 to 10 hours.

[0080] (6) The solvent was dried using a rotary evaporator to obtain red solid particles. The crude product was then dissolved in EtOAc (50 ml) and poured into a separatory funnel. The mixture was washed with water (100 ml × 3) to remove water-soluble impurities. The resulting organic layer was dried over anhydrous NaSO and filtered. The filtrate was again dried using a rotary evaporator to remove the solvent. The resulting crude product was dried in a vacuum drying oven at room temperature to obtain the HNO-releasing compound described in this example.

[0081] Example 3

[0082] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0083]

[0084] Wherein R is the following group:

[0085]

[0086] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is 2-pyridinecarbonyl chloride.

[0087] Example 4

[0088] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0089]

[0090] Wherein R is the following group:

[0091]

[0092] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is p-fluorobenzoyl chloride.

[0093] Example 5

[0094] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0095]

[0096] Wherein R is the following group:

[0097]

[0098] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is p-tert-butylbenzoyl chloride.

[0099] Example 6

[0100] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0101]

[0102] Wherein R is the following group:

[0103]

[0104] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is m-chlorobenzoyl chloride.

[0105] Example 7

[0106] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0107]

[0108] Wherein R is the following group:

[0109]

[0110] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the selection of the acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is quinoline-2-carbonyl chloride.

[0111] Example 8

[0112] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0113]

[0114] Wherein R is the following group:

[0115]

[0116] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is 3,5-difluorobenzoyl chloride.

[0117] Example 9

[0118] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0119]

[0120] Wherein R is the following group:

[0121]

[0122] The preparation method of the HNO-releasing compound described in this example is the same as that in Example 2, except for the different acylating agent used. In the preparation method of the HNO-releasing compound described in this example, the acylating agent is methyl 4-chloroformylbenzoate.

[0123] Example 10

[0124] An embodiment of the compound capable of releasing HNO of the present invention, the structural formula of the compound capable of releasing HNO of this embodiment is as follows:

[0125]

[0126] Wherein R is the following group:

[0127]

[0128] The method for preparing the HNO-releasing compound described in this example is identical to that of Example 2, except for the selection of the acylating agent used. In the method for preparing the HNO-releasing compound described in this example, the acylating agent is 2-(4-chlorophenyl)-3-methylbutyryl chloride.

[0129] Example 11

[0130] The H NMR spectrum, C NMR spectrum, mass spectrum and kinetic degradation data of the compound described in Example 1 and the H NMR spectrum and kinetic degradation data of the compound described in Example 2 were detected respectively. The detection method is:

[0131] H NMR spectrum: 2.0 mg of the compound was dissolved in 0.7 mL of deuterated dimethyl sulfoxide, the resulting solution was added to an NMR tube, and sent to a NMR detection center for detection. The results of the compound detection in Example 1 are shown in the attached figure. Figure 3 As shown. Figure 3 It can be seen that the inventors of the present application synthesized the target product having the structure described in Example 1 with high purity.

[0132] NMR carbon spectrum: 20 mg of the compound was dissolved in 0.7 mL of deuterated dimethyl sulfoxide, the resulting solution was added to an NMR tube, and sent to a NMR detection center for detection. The results of the compound detection in Example 1 are shown in the attached figure. Figure 4 As shown. Figure 4 It can be seen that the inventor of this application has Figure 1 The synthetic route can obtain the compound of the structure described in Example 1 of the present application with high purity.

[0133] Mass spectrometry: 1 mg of the compound was dissolved in 10 mL of anhydrous methanol, and 0.1 mL of the resulting solution was diluted in 10 mL of anhydrous methanol and sent to a mass spectrometry detection center for detection. The detection results of the compound in Example 1 are shown in the attached figure. Figure 5 As shown. Figure 5 It can be seen that the molecular weight of the compound of Example 1 detected by mass spectrometry is consistent with the molecular weight calculated by the inventors of the present application.

[0134] Kinetic degradation data: The test conditions are as follows: the compound of Example 1 (20 μM) was added to a cuvette at a volume ratio of 5:95 (MeCN: phosphate buffer 0.1 M, pH 7.4) at 37°C. The cuvette was quickly placed in the sample well of a UV-visible spectrometer for testing. The parameters were set to measure once every 15 minutes. The reaction rate constant obtained for the compound of Example 1 was 0.01019±8.4077×10 -4 s -1 , reaction half period t 1 / 2 The test results are as follows: 68 minutes Figure 6 The degradation kinetics of the compound of Example 2 at pH 12 was 7 min, and the degradation kinetics at pH 10 was 120 min.

[0135] By the attached Figure 6 It can be seen that there is an obvious uniform and regular degradation trend at the wavelength of 450-550nm, which means that the compound with the structure described in Example 1 of the present application is used. The compound can be homogeneously degraded under physiological conditions. Figure 6As can be seen from the illustration, the nonlinear fitting at 476 nm shows that the degradation rate of the compound described in the examples of the present application is uniform and slow, with trend changes, and also shows a trend of first-order reaction. Where: K = 0.010198.40877 × 10 -4 , t 1 / 2 =68min.

[0136] The hydrogen nuclear magnetic resonance spectrum of the compound of Example 2 is: 1H NMR (400 MHz, DMSO-d6) δ9.89 (s, 1H), 7.99 (dd, J = 14, 8.8, 400 Hz, 4H), 7.89 (d, J = 9.2 Hz, 2H), 7.88 (d, J = 9.2, 400 Hz, 2H), 3.50 (s, 6H), 3.79 (s, 3H).

[0137] The detection methods and results of the compounds described in Examples 3 to 10, such as H NMR spectrum, C NMR spectrum, mass spectrum and kinetic degradation data, are similar to those of the compounds described in Examples 1 and 2 and are not described in detail here.

[0138] Example 12

[0139] Detection of HNO production standards

[0140] Using phosphorus-containing compounds to react with the compound of Example 1 31 P NMR detection

[0141] The phosphorus-containing compound (methyl-2-diphenylphosphine terephthalate) was reacted with a pH 10.2 buffer solution, the compound of Example 1, and a mixture of deuterated acetonitrile (CD3CN). 31 P NMR confirmed the formation of HNO. The results are shown in the attached Figure 7 shown.

[0142] By the attached Figure 7 It can be seen that 31 There are three obvious single peaks in the P NMR spectrum. Through the design of substance concentration, it can be seen from the integration that their ratio is 1:1:1, phosphorus oxide 2 (δ33.88ppm) and phosphoramide 3 (δ34.53ppm). Therefore, it can be confirmed that HNO reacts completely with phosphorus-containing compounds and HNO is 31 P NMR captures and also confirms that the compound of Example 1 described in the present application can release HNO.

[0143] The compounds described in Examples 2 to 10 were tested using the same method, and all of them were able to demonstrate the generation of HNO, which will not be repeated here.

[0144] Example 13

[0145] Fluorescence property detection of the compound capable of releasing HNO according to the present invention

[0146] The compound described in Example 1 of the present invention was used as the test object. The compound of Example 1 (50 μM) was mixed with a volume ratio of 5:95 (MeCN: phosphate buffer solution 0.1M, pH 7.4). The fluorescence performance was tested at a temperature of 25 degrees. The degraded compound was prepared in advance and placed in a 50-degree oven overnight. The test results are shown in the attached figure. Figure 8 As shown. Figure 8 It can be seen that the fluorescence intensity of the compound described in Example 1 of the present application before and after degradation is compared at the excitation wavelength.

[0147] By the attached Figures 3 to 8 The results show that the HNO-releasing compound described in Example 1 of the present application can be degraded and release HNO under physiological conditions and alkaline conditions, and its release half-cycle ranges from several minutes to several hours. At the same time, the degradation product is a fluorescent product.

[0148] The compounds described in Examples 2 to 10 were tested using the same method and had similar effects to the compound in Example 1, which will not be described in detail here.

[0149] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Use of a compound capable of releasing HNO in the preparation of a drug for treating heart failure and cardiovascular diseases, characterized in that: The structural formula of the compound is as follows: Where R is H or 2. A method for preparing a compound capable of releasing HNO, characterized in that: The structural formula of the compound is as follows: Where R is H or The preparation method comprises the following steps: (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution. (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution; (3) drying the reaction solvent of the light red reaction solution obtained in step (2) by a rotary evaporator, and then drying the obtained initial product in a vacuum drying oven at room temperature for 24 to 48 hours to obtain the compound capable of releasing HNO; or (1) Add NH2OH·HCl and triethylamine to a reaction flask, stir evenly, then add tetrahydrofuran and continue stirring until white turbidity appears in the reaction flask. Then weigh 4-dimethylaminophenylazobenzenesulfonyl chloride and slowly add it to the reaction flask. The solution in the reaction flask turns red. React at room temperature for 12 to 20 hours to obtain a red reaction solution. (2) EtOAc was added to the reaction flask of step (1), and the mixture was washed and extracted with water. The obtained organic phase was dried over anhydrous Na2SO4, and the Na2SO4 solid was removed by suction filtration to obtain a light red reaction solution; (3) The light red reaction solution obtained in step (2) was dried using a rotary evaporator to dry the reaction solvent, and the obtained primary product was dried in a vacuum drying oven at room temperature for 24 to 48 hours to obtain an intermediate; (4) Under N2 atmosphere and ice-water bath conditions, weigh the intermediate obtained in step (3) and pour it into a reaction flask, and add anhydrous THF to dissolve it; (5) Add triethylamine to the reaction flask, then slowly add the acylating agent to the reaction flask. After 10 to 40 minutes, remove the ice-water bath and stir the reaction at room temperature for 5 to 10 hours. (6) The solvent was dried using a rotary evaporator to obtain red solid particles, and then the crude product was dissolved in EtOAc, poured into a separatory funnel, and washed with water to remove water-soluble impurities. The obtained organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was again dried using a rotary evaporator to dry the solvent. The obtained preliminary product was dried in a vacuum drying oven at room temperature to obtain the compound capable of releasing HNO; The acylating agent is acetyl chloride.

3. The method for preparing a compound capable of releasing HNO as claimed in claim 2, wherein: The molar ratio of NH2OH·HCl to 4-dimethylaminophenylazobenzenesulfonyl chloride in step (1) is: NH2OH·HCl:4-dimethylaminophenylazobenzenesulfonyl chloride=2.5-6.0:

1.

4. The method for preparing a compound capable of releasing HNO as claimed in claim 2, wherein: In step (1), at least one of the following conditions is met: (a) the molar ratio of NH2OH·HCl to triethylamine is: NH2OH·HCl:triethylamine=3.2-5:1 mmol / ml; (b) The molar volume ratio of NH2OH·HCl to tetrahydrofuran is: NH2OH·HCl:tetrahydrofuran = 1.01-1.6:1 mmol / ml.

5. The method for preparing a compound capable of releasing HNO as claimed in claim 2, wherein: In step (5), at least one of the following conditions is met: (a) the molar ratio of the acylating agent to the intermediate is: acylating agent: intermediate = 1.1 to 1.3:1; (b) The molar ratio of triethylamine to the intermediate is: triethylamine:intermediate=1.1-1.3:

1.

6. The method for preparing a compound capable of releasing HNO according to claim 4 or 5, wherein: At least one of the following must be met: (1) The molar ratio of NH2OH·HCl to triethylamine in step (1) is: NH2OH·HCl:triethylamine=15.576:3.406; (2) The molar volume ratio of NH2OH·HCl to tetrahydrofuran in step (1) is: NH2OH·HCl:tetrahydrofuran = 15.576:10 mmol / ml; (3) The molar ratio of the acylating agent to the intermediate in step (5) is: acylating agent:intermediate=0.343:0.312; (4) The molar ratio of triethylamine to the intermediate in step (5) is: triethylamine:intermediate=0.343:0.312.

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