Preparation of 4-hydroxythiobenzamide hydrogen sulfide donor derivatives without catalyst based on onium methanesulfonate

Abstract

This invention provides a catalyst-free preparation method for 4-hydroxythiobenzamide hydrogen sulfide donor derivatives based on ononium methanesulfonate, belonging to the field of medicinal chemistry. This TBZ-type hydrogen sulfide donor derivative has the structure shown in Formula I: Formula I. This invention converts 4-hydroxythiobenzamide into methanesulfonate form (TBZ-type hydrogen sulfide donor derivative), and when coupled with other drug molecules, the reaction is convenient, readily available, catalyst-free, and produces products with good stability.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a catalyst-free method for preparing 4-hydroxythiobenzamide hydrogen sulfide donor derivatives based on ononium methanesulfonate. Background Technology

[0002] Hydrogen sulfide (H2S) is a colorless and irritating gas. Studies have found that it is not only endogenously synthesized in mammals but also participates in various physiological and pathological processes. It is the third signal transduction molecule discovered in living organisms after carbon monoxide (CO) and nitric oxide (NO). At appropriate concentrations, H2S not only possesses anti-inflammatory, anti-cancer, and antioxidant biological activities but also participates in immune and metabolic regulation, showing therapeutic effects in diseases such as myocardial infarction and diabetes.

[0003] Numerous studies have shown that H2S concentrations in the body are lower than normal physiological levels during disease states, suggesting that maintaining stable H2S concentrations may be an effective strategy for disease prevention and intervention. H2S donors can convert gaseous H2S into a stable solid form, effectively addressing its volatility and difficulty in regulation. Currently, exogenous H2S donors are mainly divided into two types: inorganic and organic. Inorganic H2S donors primarily include Na2S and NaSH, but they have drawbacks such as rapid release over a short period leading to excessively high concentrations and toxicity, and difficulty in mimicking the body's continuous and slow endogenous production of H2S. Therefore, current research mainly focuses on organic H2S donors.

[0004] Considering the therapeutic effects of metabolic stability and slow release on diseases, current research mainly focuses on linking H2S donors with existing drugs to obtain H2S donor conjugates for disease treatment. Representative examples include a series of H2S donor conjugates synthesized by conjugating H2S donors with nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, naproxen, diclofenac, and ketoprofen. These conjugates not only enhance the anti-inflammatory activity of NSAIDs but also reduce or even counteract the gastrointestinal side effects caused by NSAID monomers. For instance, Luo Bilan et al. synthesized H2S donor conjugates by combining tetramethylpyrazine (TMP) with the H2S donor 5-(4-hydroxyphenyl)-3H-1,2-dithiacyclopenten-3-thionone (ADT-OH), which showed in vitro efficacy against AD. P-induced platelet aggregation showed superior inhibitory activity compared to the lead compound tetramethylpyrazine, and one compound even outperformed commonly used clinical drugs butylphthalide, aspirin, and edaravone, making it a promising candidate compound for cardiovascular and cerebrovascular diseases. For example, Hu et al. synthesized an H2O2-responsive doxorubicin (DOX) conjugated prodrug that specifically releases DOX and H2S gas upon H2O2 triggering. This prodrug still exhibited significant toxicity to DOX-sensitive triple-negative breast cancer cells in mice, and reduced cardiotoxicity while retaining antitumor activity, demonstrating potential for clinical translation.

[0005] 4-hydroxythiobenzamide (TBZ) is a type of H2S donor. The phenol at position 4 in its structure is a traditional linker. If you want to couple it with other drug molecules, current synthetic methods require expensive condensing agents, catalysts, or reactions under strong alkaline conditions. Summary of the Invention

[0006] In view of this, the purpose of this invention is to provide a catalyst-free preparation method for 4-hydroxythiobenzamide hydrogen sulfide donor derivatives based on ononium methanesulfonate. This invention converts 4-hydroxythiobenzamide into the form of methanesulfonate (TBZ type hydrogen sulfide donor derivative), and then couples it with other drug molecules. This reaction has the advantages of being convenient and readily available, catalyst-free, and having good product stability.

[0007] To achieve the above objectives, the present invention provides the following technical solution: One of the technical solutions of this invention is a TBZ-type hydrogen sulfide donor derivative having the structure shown in Formula I: Formula I.

[0008] The second technical solution of the present invention is a method for preparing the above-mentioned TBZ-type hydrogen sulfide donor derivative, comprising the following steps: In organic solvent 1, 1,1'-carbonyldiimidazole and 4-hydroxythiobenzamide were mixed and reacted to obtain 4-thiocarbamoylbenzene 1H-imidazol-1-carboxylate. In organic solvent 2, the 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate and methanesulfonic acid are mixed and reacted to obtain the TBZ-type hydrogen sulfide donor derivative.

[0009] The third technical solution of this invention is a method for preparing an H2S donor conjugate. When the drug is mexiletine hydrochloride, the preparation method includes the following steps: In organic solvent 3, mexiletine hydrochloride was mixed with a compound with the structure shown in Formula I and reacted to obtain product compound II. The structural formula of compound II is: ; When the drug is propranolol hydrochloride, the preparation method includes the following steps: (1) In an alkaline aqueous system, propranolol hydrochloride and ditert-butyl dicarbonate were reacted to obtain compound III; (2) In dichloromethane, N-fluorenylmethoxycarbonyl-β-alanine, 4-dimethylaminopyridine (DMAP), compound III, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were reacted for 5 to obtain compound IV; (3) Compound IV and piperidine were reacted in ethyl acetate to give compound V; (4) In DMF, the compound shown in Formula I and compound V are reacted to obtain compound VI; (5) After mixing compound VI and TFA in DCM solution, reaction 8 was carried out to obtain target compound VII; The structural formula of compound III is: ; The structural formula of compound IV is: ; The structural formula of compound V is: ; The structural formula of compound VI is: ; The structural formula of compound VII is: .

[0010] The present invention discloses the following technical effects: This invention converts 4-hydroxythiobenzamide into a methanesulfonate form (TBZ-type hydrogen sulfide donor derivative), and then couples it with other drug molecules. This reaction has the advantages of being convenient and readily available, having no catalyst dependence, and producing products with good stability.

[0011] 4-hydroxythiobenzamide is converted into a methanesulfonate form (TBZ-type hydrogen sulfide donor derivative), and then coupled with other drug molecules. The resulting compound can slowly release hydrogen sulfide into cells to produce a therapeutic effect. Attached Figure Description

[0012] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0013] Figure 1 This is the FTIR spectrum of compound I in Example 1 of the present invention.

[0014] Figure 2 This is the general reaction formula for compound A in Example 1.

[0015] Figure 3 The figure shows the CCK8 cell viability results of compound VII under a cell hypoxia-reoxygenation model.

[0016] Figure 4 This is a fluorescence image of compound VII releasing hydrogen sulfide in living cells.

[0017] Figure 5 The calibration curve for Na2S in Example 4 is shown.

[0018] Figure 6 The amount of hydrogen sulfide released by different compounds. Detailed Implementation

[0019] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0020] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0021] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0022] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0023] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0024] In this invention, room temperature refers to 15~30℃.

[0025] The first aspect of this invention provides a TBZ-type hydrogen sulfide donor derivative having the structure shown in Formula I: Formula I.

[0026] The method for preparing the TBZ-type hydrogen sulfide donor derivative with the structure shown in Formula I includes the following steps: In organic solvent 1, 1,1'-carbonyldiimidazole and 4-hydroxythiobenzamide were mixed and reacted to obtain 4-thiocarbamoylbenzene 1H-imidazol-1-carboxylate. In organic solvent 2, the 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate ester and methanesulfonic acid are mixed and reacted to obtain the TBZ-type hydrogen sulfide donor derivative. The general reaction formula is: .

[0027] Specifically: 1,1'-carbonyldiimidazole is dissolved in a first organic solvent to obtain a first solution; 4-hydroxythiobenzamide is dissolved in a second organic solvent to obtain a second solution; The second solution was added dropwise to the first solution to carry out reaction 1, yielding 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate; The 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate ester was dissolved in a third organic solvent to obtain a third solution; Methanesulfonic acid was added dropwise to the third solution to carry out reaction 2, thereby obtaining the TBZ-type hydrogen sulfide donor derivative.

[0028] In a preferred embodiment of the present invention, the first organic solvent and the second organic solvent are the same, both being acetonitrile. The present invention does not impose any special limitation on the amount of the first organic solvent and the second organic solvent, as long as the amount is sufficient to fully dissolve the reactants and satisfy the reaction requirements.

[0029] In a preferred embodiment of the present invention, the molar ratio of 1,1'-carbonyldiimidazole to 4-hydroxythiobenzamide is (50~52):(25~26).

[0030] In a preferred embodiment of the present invention, the temperature of reaction 1 is room temperature, the reaction is monitored by thin-layer chromatography, 4-hydroxythiobenzamide disappears, and the reaction ends.

[0031] In a preferred embodiment of the present invention, after reaction 1 is completed, the process further includes a step of filtration, washing and drying the resulting filter cake; the washing agent used is ethyl acetate, and the drying method is natural air drying.

[0032] In a preferred embodiment of the present invention, the molar ratio of 4-thiocarbamoylbenzene 1H-imidazol-1-carboxylate to methanesulfonic acid is 1:(0.9~1.1).

[0033] In a preferred embodiment of the present invention, the third organic solvent is DMF. The present invention does not impose a special limitation on the amount of the third organic solvent, as long as it is sufficient to fully dissolve the reactants and allow the reaction to proceed.

[0034] In a preferred embodiment of the present invention, the temperature of reaction 2 is room temperature, the time is 2-3 hours, and the reaction is stirred.

[0035] In a preferred embodiment of the present invention, the reaction is further further comprising the steps of filtration, washing and drying the resulting filter cake; the washing agent is ethyl acetate, and the drying method is natural air drying.

[0036] A second aspect of this invention provides a method for preparing an H2S donor conjugate. When the drug is mexiletine hydrochloride, the preparation method includes the following steps: In organic solvent 3, mexiletine hydrochloride was mixed with a compound with the structure shown in Formula I and reacted to obtain product compound II. The structural formula of compound II is: .

[0037] The general reaction formula is: .

[0038] Specifically: When the compound with the structure shown in Formula I is mixed with the fourth organic solvent, a fourth solution is obtained; Dissolve mexiletine hydrochloride in the fifth organic solvent to obtain the fifth solution; The fifth solution was added dropwise to the fourth solution to carry out reaction 3, yielding product compound II.

[0039] In a preferred embodiment of the present invention, the fourth organic solvent and the fifth organic solvent are the same, both being DMF. The present invention does not impose any special limitation on the amount of the fourth and fifth organic solvents, as long as their amounts are sufficient to fully dissolve the reactants and allow the reaction to proceed.

[0040] In a preferred embodiment of the present invention, the molar ratio of mexiletine hydrochloride to the compound with the structure shown in Formula I is 1:(0.9~1.1); the reaction 3 is carried out at room temperature for 17~19 hours.

[0041] In a preferred embodiment of the present invention, after reaction 3 is completed, the process further includes extracting the organic phase, then adding anhydrous sodium sulfate to the organic phase for drying, filtering, rotary evaporation, and rapid column chromatography purification of the crude product (using dichloromethane and methanol as the developing solvent, 4% methanol).

[0042] When the drug is propranolol hydrochloride, the preparation method includes the following steps: (1) In an alkaline aqueous system, propranolol hydrochloride and ditert-butyl dicarbonate were reacted to obtain compound III; (2) In organic solvent 4, N-fluorenylmethoxycarbonyl-β-alanine, 4-dimethylaminopyridine (DMAP), compound III, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were reacted to obtain compound IV; (3) In organic solvent 5, compound IV and piperidine are reacted to obtain compound V; (4) In organic solvent 6, the compound shown in formula I and compound V are reacted to obtain compound VI; (5) After mixing compound VI and TFA in DCM solution, reaction 8 was carried out to obtain target compound VII; The structural formula of compound III is: ; The structural formula of compound IV is: ; The structural formula of compound V is: ; The structural formula of compound VI is: ; The structural formula of compound VII is: .

[0043] The general reaction formula is: .

[0044] In a preferred embodiment of the present invention, in step (1), the alkaline aqueous phase system is a sodium bicarbonate solution; the molar ratio of propranolol hydrochloride and ditert-butyl dicarbonate is 1:(1.1~1.3), the reaction temperature is room temperature, and the reaction is monitored by TLC.

[0045] In a preferred embodiment of the present invention, after reaction 4 is completed, the process further includes the steps of extracting the system with ethyl acetate, collecting the organic phase, and then drying, filtering, and evaporating the organic phase.

[0046] In a preferred embodiment of the present invention, in step 2, the molar ratio of N-fluorenemethoxycarbonyl-β-alanine, 4-dimethylaminopyridine, compound III, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride is (1.0~1.2):(4~5):1.0:(2~3); the reaction temperature in step 5 is room temperature, and the reaction is monitored by TLC.

[0047] In a preferred embodiment of the present invention, the organic solvent 4 is dichloromethane.

[0048] In a preferred embodiment of the present invention, after reaction 5 is completed, the process further includes adding HCl to the system to quench the reaction, then adjusting the pH to <3, extracting, drying the organic phase with anhydrous sodium sulfate, filtering, evaporating under reduced pressure, purifying by column chromatography with a V(petroleum ether):V(ethyl acetate) ratio of 60:40, and evaporating under reduced pressure.

[0049] In a preferred embodiment of the present invention, in step (3), the molar ratio of compound IV to piperidine is 1:(4~6), the reaction temperature is room temperature, and the reaction is monitored by TLC.

[0050] In a preferred embodiment of the present invention, the organic solvent 5 is ethyl acetate.

[0051] In a preferred embodiment of the present invention, after reaction 6 is completed, the process further includes the steps of sequentially removing the solvent under reduced pressure, purifying by column chromatography with a ratio of V(dichloromethane):V(methanol) of 95:5, and then evaporating under reduced pressure.

[0052] In a preferred embodiment of the present invention, in step (4), the molar ratio of the compound shown in formula I to compound V is (1.5~2):(2~2.5), the temperature of reaction 7 is 50°C, and the time is 4h.

[0053] In a preferred embodiment of the present invention, the organic solvent 6 is DMF.

[0054] In a preferred embodiment of the present invention, after reaction 7 is completed, the reaction solution is further dissolved in ethyl acetate, extracted with water, then extracted with saturated sodium chloride, the organic phase is dried with anhydrous sodium sulfate, filtered, evaporated under reduced pressure, and the crude product is purified by rapid column chromatography [V(petroleum ether):V(ethyl acetate)=50:50].

[0055] In step (5), the molar ratio of compound VI to TFA is 1:(5-10); the reaction 8 is carried out under ice bath conditions and the reaction is monitored by TLC.

[0056] In a preferred embodiment of the present invention, after reaction 8 is completed, the process further includes the steps of decompressing the solvent under reduced pressure, adding ethyl acetate and decompressing to remove TFA, followed by column chromatography purification, dry loading, preparing V(dichloromethane):V(methanol) = 98:2 under medium pressure, a second dry mixing, preparing D:M (5% M product) under medium pressure, and decompressing.

[0057] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.

[0058] To better understand the present invention, the following embodiments further illustrate the content of the present invention, but the content of the present invention is not limited to the following embodiments.

[0059] Example 1 1,1'-carbonyldiimidazole (51.18 mmol) was dissolved in 80 mL of ultra-dry acetonitrile by sonication in a 500 mL round-bottom flask. Separately, 25.59 mmol of 4-hydroxythiobenzamide was dissolved in 120 mL of ultra-dry acetonitrile by sonication and added to a constant-pressure dropping funnel. This solution was slowly added dropwise to the round-bottom flask over 0.5 h with continuous stirring at room temperature. The reaction was monitored by thin-layer chromatography. The reaction was considered complete when the 4-hydroxythiobenzamide disappeared. After the reaction was complete, the mixture was filtered, and the filter cake was washed with 100 mL of cold ethyl acetate and dried to obtain 6.15 g of a yellow powder, namely the product 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate, with a yield of 97.23% and a melting point of 180.5-181.5℃. 1 H NMR (400 MHz, DMSO) δ 10.00 (s, 1H), 9.61 (s, 1H), 8.50 (d,J = 4.1 Hz, 1H), 7.99 (d, J = 8.7 Hz, 2H), 7.80 (d, J = 4.1 Hz, 1H), 7.49 (d,J = 8.6 Hz, 2H), 7.17 (s, 1H); 13 C NMR (101 MHz, DMSO) δ 199.37, 152.18, 147.24, 138.53, 138.37, 131.09, 129.39, 121.58, 118.51. HR-MS, m / Z :C 11 H 10 N3O2S [M+H] + Theoretical value: 248.0488; measured value: 248.0494.

[0060] Weigh 32.35 mmol of 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate into a 250 mL reaction flask and sonicate it into 80 mL of ultra-dry DMF. Slowly add 32.35 mmol of methanesulfonic acid (DMSA) dropwise to the DMF solution of 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate over 30 minutes, and stir continuously at room temperature for 3 h. After the reaction is complete, filter the solution. Wash the filter cake with 150 mL of cold ethyl acetate and air dry to obtain 8.6 g of a pale yellow powder, which is the product 1-((4-thiocarbamoylphenoxy)carbonyl)-1H-imidazolium-3-onium methanesulfonate (compound I). Yield: 77.41%, melting point: 182.1–183.0 °C.

[0061] Because compound I has an onium methanesulfonate structure, its solubility is extremely poor. Conventional mass spectrometry analysis is insufficient to obtain an effective signal. Therefore, its structure was indirectly confirmed by infrared spectroscopy characterization and derivatization with methanol to generate downstream product (A).

[0062] The FTIR spectrum of compound I is as follows: Figure 1 As shown. By Figure 1 It can be known that 3301.4 cm -1 Corresponding peak for the NH stretching vibration of thioamide; 3156 cm⁻¹ -1 Corresponding CH stretching vibration peaks for the benzene and imidazole rings; 1804.2 cm⁻¹ -1 The wavenumber of the C=O stretching vibration peak corresponding to the ester group is higher than that of the conventional ester carbonyl group (~1735 cm⁻¹). -1 The enhanced electron-deficient effect of the imidazole ring forming a salt with the sulfonate group increases the polarity of the C=O bond and the force constant, resulting in a characteristic blue shift in the absorption peaks. This phenomenon strongly supports the existence of this unique structure; 1654.4, 1594.4 cm⁻¹ -1 The skeletal stretching vibration peak corresponding to the episial disubstituted benzene ring is 829.1 cm⁻¹. -1 The out-of-plane bending vibration peak corresponding to the type of benzene ring substitution is 1351.2 cm⁻¹. -1 With 1224.9 cm -1 Corresponding to sulfonate (-SO3) - The asymmetric and symmetric stretching vibration peaks of the instrument; 1272.5 cm⁻¹ -1 Corresponding to the C=S stretching vibration peak, 1148.6 cm⁻¹ -1 Corresponding CO stretching vibration peaks of the ester group; 1035.8, 895.8 cm⁻¹ -1 The isofinite absorption peak is a unique characteristic peak resulting from the combined effects of intramolecular functional groups, benzene ring and imidazole ring conjugation, and steric effects. In summary, the infrared spectral characteristic absorption matches the functional groups and electronic environment of compound I, which can verify its structure.

[0063] The specific reaction steps for product A are as follows: In a 50 mL round-bottom flask, compound I (0.5 g, 1.46 mmol) was added, followed by excess methanol, and the mixture was stirred continuously at room temperature for 17 h. After the reaction was complete, the reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by rapid column chromatography under medium pressure [V(petroleum ether):V(ethyl acetate) = 13:7], yielding 251 mg of a yellow solid, namely the product methyl 4-thiocarbamoylphenyl carbonate (A), with a yield of 81.6% and a melting point of 183.5–184.5 °C. ¹H NMR (400 MHz, DMSO) δ 9.94 (s, 1H), 9.56 (s, 1H), 7.99–7.88 (m, 2H), 7.35–7.23 (m, 2H), 3.84 (s, 3H); ¹³C NMR (101 MHz, DMSO) δ 199.40, 153.72, 153.22, 137.84, 129.32, 121.20, 56.10.HR-MS, m / Z: C9H9NO3S [M+H]+ Theoretical value 212.0376, measured value 212.0372.

[0064] Compound A like Figure 2 In reaction one, compound I reacts with methanol under the above reaction conditions to obtain product A in 81.6% yield without a basic catalyst. However, in reaction two, when 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate reacts with methanol under the same reaction conditions, almost no product is formed.

[0065] The above facts demonstrate that 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate reacts with methanesulfonic acid to form a novel onium salt, whose structure and reactivity differ from those of 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate. This is indirect evidence. Due to its extremely poor solubility, compound I cannot be detected by mass spectrometry and nuclear magnetic resonance.

[0066] Example 2 Mexiletine hydrochloride was extracted with saturated sodium bicarbonate solution and ethyl acetate to obtain a colorless oily mexiletine. Compound I (2.91 mmol) was weighed into a clean, dry 50 mL flask, and 12 mL of ultra-dry DMF was added. Mexiletine (2.91 mmol) was weighed and dissolved in 3 mL of ultra-dry DMF, then added dropwise to the flask. The mixture was stirred continuously at room temperature for 17 h. After the reaction was complete, the mixture was extracted with water and ethyl acetate. The organic phase was dried with anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by rapid column chromatography (using dichloromethane and methanol as the developing solvent, 4% methanol) to give 154 mg of a yellow solid, namely the product 4-thiocarbamoylphenyl N-(1-(2,6-dimethylphenoxy)propyl-2-yl)carbamate (compound II), with a yield of 14.75% at 100.5–101.5 °C. 1 H NMR (400 MHz, DMSO) δ 9.88 (s, 1H), 9.51 (s,1H), 8.05 (d, J = 8.1 Hz, 1H), 7.98 – 7.88 (m, 2H), 7.17 – 7.12 (m, 2H), 7.02(d, J = 7.5 Hz, 2H), 6.91 (dd, J = 8.2, 6.8 Hz, 1H), 4.00 – 3.90 (m, 1H), 3.71 (d, J = 5.5 Hz, 2H), 2.24 (s, 6H), 1.30 (d, J = 6.8 Hz, 3H); 13 C NMR (101MHz, DMSO) δ 199.53, 170.82, 155.52, 153.82, 136.58, 130.84, 130.80, 129.25,129.22, 129.15, 124.25, 124.16, 121.45, 74.30, 60.24, 47.66, 39.33, 17.61,16.35, 14.56; HR-MS, m / z, C 19 H 22 N₂O₃S [M+H] + Theoretical value: 359.1424, measured value: 359.1429.

[0067] Example 3 (1) Compound III Take a 250 mL round-bottom flask, add sodium bicarbonate (3.08 g, 36.44 mmol, 2.2 eq), and dissolve it in 80 mL of water. Then add propranolol hydrochloride (5 g, 16.56 mmol, 1.0 eq). After the system is clear, add di-tert-butyl dicarbonate (4.66 mL, 19.88 mmol, 1.2 eq) dropwise. Stir at room temperature for 18 h. Monitor the reaction by TLC. After the reaction is complete, transfer the system to a separatory funnel and extract with ethyl acetate. Combine the organic phases, dry with anhydrous sodium sulfate, filter, evaporate to dryness, and then vacuum to obtain 5.87 g of white solid compound III, yield 94.81%. 1 H NMR (400 MHz, CDCl3) δ 8.35– 8.00 (m, 1H), 7.90 – 7.70 (m, 1H), 7.57 – 7.41 (m, 3H), 7.38 (t, J = 7.9Hz, 1H), 6.85 (d, J = 7.5 Hz, 1H), 4.31 – 3.98 (m, 4H), 3.53 (d, J = 5.2 Hz, 2H), 1.51 (s, 9H), 1.25 (d, J = 6.9 Hz, 3H), 1.16 (d, J = 6.7 Hz, 3H); 13 C NMR(101 MHz, CDCl3) δ 154.17, 134.52, 127.62, 126.43, 125.93, 125.46, 125.27,121.68, 120.62, 104.79, 80.79, 69.87, 48.70, 47.11, 28.50, 27.44, 21.04,20.52. HR-MS, m / Z: C 21 H 29 NO4[M+H] + Theoretical value: 360.2169, measured value: 360.2205.

[0068] (2) Compound IV Take a 500 mL round-bottom flask, add N-fluorenylmethoxycarbonyl-β-alanine (4.76 g, 15.3 mmol, 1.1 eq), and dissolve it in 200 mL ultra-dry dichloromethane. Then add DMAP (4.4 g, 61.2 mmol, 4.4 eq), and after it dissolves completely, place the system in an ice-salt bath. Add compound III (5 g, 13.91 mmol, 1.0 eq), then add EDCI (5.93 g, 30.6 mmol, 2.2 eq), replace the gas with nitrogen for protection, stir in the ice-salt bath for 1 h, then remove the ice bath and stir at room temperature for 14 h. The reaction was monitored by TLC. Upon completion, 0.1 M HCl was added to quench the reaction, and 0.3 M HCl was added to adjust the pH to <3. Extraction was performed, and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, evaporated under reduced pressure, and purified by column chromatography under medium pressure (V(petroleum ether):V(ethyl acetate) = 60:40). Evaporation under reduced pressure yielded 8.096 g of a pale yellow oily liquid compound IV, with a yield of 89.16%. 1 H NMR(400 MHz, CDCl3) δ 8.11 (dd, J = 6.5, 3.3 Hz, 1H), 7.66 (dd, J = 14.7, 8.6Hz, 3H), 7.46 – 7.11 (m, 11H), 6.69 (d, J = 7.6 Hz, 1H), 5.54 (s, 2H), 4.18 (p, J = 8.0 Hz, 4H), 4.02 (d, J = 7.0 Hz, 2H), 3.48 (s, 1H), 3.39 (d, J = 6.4Hz, 3H), 1.39 (s, 9H), 1.10 (dd, J = 10.9, 6.7 Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 171.95, 156.37, 154.13, 143.95, 141.27, 134.52, 127.67, 127.58,127.04, 126.57, 125.78, 125.53, 125.49, 125.12, 121.73, 120.92, 119.96,104.78, 80.20, 67.97, 66.82, 47.16, 36.63, 34.77, 28.51, 21.04. HR-MS, m / Z:C 39 H 44 N₂O₇[M+H] + Theoretical value: 653.3227; measured value: 653.3198.

[0069] (3) Compound V Take a 250 mL round-bottom flask, add compound IV (7.96 g, 12.19 mmol, 1 eq), dissolve in 55 mL ethyl acetate, and add piperidine (6.02 mL, 60.94 mmol, 5.0 eq) dropwise. Under nitrogen protection, purge the mixture and stir at room temperature for 2 h. Monitor the reaction by TLC. After the reaction is complete, remove the solvent under reduced pressure and purify by column chromatography under medium pressure. The solution is V(dichloromethane):V(methanol) = 95:5. The solution is evaporated to dryness under reduced pressure to give 3.2287 g of white solid compound V, yield 61.53%. HR-MS, m / Z:C 24 H 34 N₂O₅[M+H] + The theoretical value is 431.2546, and the measured value is 431.2244. The product is unstable and easily decomposes, so it is immediately used in the next reaction. Therefore, there is no NMR data.

[0070] (4) Compound VI Weigh 0.6 g (1.75 mmol) of 1-((4-thiocarbamoylphenoxy)carbonyl)-1H-imidazolium-3-onium methanesulfonate (compound I) in a 50 mL round-bottom flask, add 5 mL of anhydrous DMF, and separately dissolve compound V (1.05 g, 2.45 mmol) in 5 mL of anhydrous DMF and add it dropwise to the flask. Stir continuously at 50 °C for 4 h. After the reaction is complete, dissolve the reaction solution in 40 mL of ethyl acetate, extract with water (100 mL × 2), and then extract with 10 mL of saturated sodium chloride. Dry the organic phase with anhydrous sodium sulfate, filter, evaporate to dryness under reduced pressure, and purify the crude product by rapid column chromatography [V(petroleum ether):V(ethyl acetate) = 50:50] to obtain 0.437 g of pale yellow solid, namely product compound VI, with a yield of 41.02% and a melting point of 75-76 °C. 1H NMR (400 MHz, DMSO) δ10.38 (s, 1H), 10.01 (s, 1H), 8.69 – 8.62 (m, 1H), 8.47 (t, J = 5.6 Hz, 1H), 8.39 (dd, J = 9.0, 6.6 Hz, 3H), 8.07 – 7.97 (m, 3H), 7.92 (t, J = 7.9 Hz, 1H), 7.62 – 7.56 (m, 2H), 7.46 (d, J = 7.6 Hz, 1H), 4.86 – 4.72 (m, 2H), 4.54(q, J = 7.1 Hz, 1H), 4.02 (d, J = 4.4 Hz, 1H), 3.89 – 3.83 (m, 2H), 3.11 (t,J = 7.0 Hz, 2H), 3.01 (p, J = 1.8 Hz, 2H), 1.91 (s, 9H), 1.65 (dt, J = 12.9,6.7 Hz, 6H); 13 C NMR (101 MHz, DMSO) δ 199.48, 171.12, 154.22, 154.06, 153.72,136.52, 134.49, 129.09, 127.96, 127.01, 126.58, 125.94, 125.33, 121.79,121.33, 120.85, 105.64, 79.47, 68.12, 60.23, 55.39, 37.01, 34.46, 28.51,21.24, 14.56. HR-MS, m / Z:C 32 H 39 N3O7S [M+H] + Theoretical value: 610.2509; measured value: 610.2593.

[0071] (5) Compound VII Take a 50 mL reaction flask, weigh compound VI (0.27 g, 0.44 mmol, 1.0 eq), and slowly add 0.41 mL of 80% TFA in DCM solution under ice bath conditions. Stir continuously for 1 h, monitor the reaction by TLC, and determine the reaction is complete. Reduce the solvent under reduced pressure, add ethyl acetate and evaporate three times (to remove TFA), purify by column chromatography, load the sample dry, and prepare a D:M ratio (V(dichloromethane):V(methanol) = 98:2) under medium pressure. Mix the sample dry for the second time, and prepare a D:M ratio (5% M yielding the product) under medium pressure. Evaporate to obtain a pale yellow solid, i.e., 0.222 g of target compound VII, yield 79.66%. 1 H NMR (400 MHz, DMSO) δ 9.87 (s, 1H), 9.52(s, 1H), 8.23 ​​– 8.12 (m, 1H), 8.02 (t, J = 5.6 Hz, 1H), 7.89 (d, J = 8.9 Hz,3H), 7.52 (td, J = 6.8, 3.3 Hz, 3H), 7.41 (t, J = 7.9 Hz, 1H), 7.08 (d, J =8.7 Hz, 2H), 6.96 (d, J = 7.6 Hz, 1H), 5.58 (dd, J = 7.5, 4.4 Hz, 1H), 4.45 –4.28 (m, 2H), 3.44 (dd, J = 47.0, 6.2 Hz, 6H), 2.77 – 2.57 (m, 2H), 1.27 (d,J = 6.5 Hz, 6H); 13 C NMR (101 MHz, DMSO) δ 199.46, 171.25, 154.33, 153.81,153.71, 136.55, 134.48, 129.11, 127.97, 127.07, 126.54, 125.99, 125.23,121.92, 121.74, 121.34, 121.09, 105.81, 68.64, 50.80, 44.66, 36.88, 34.49,19.08, 18.79. HR-MS, m / Z:C 27 H 31 N3O5S [M+H] + Theoretical value: 510.2063; measured value: 510.1495.

[0072] Efficacy validation: Bioactivity data (CCK8 immunofluorescence) 1. To evaluate the therapeutic effect of compound VII, a cardiomyocyte hypoxia-reoxygenation model was used to simulate ischemia-reperfusion injury caused by myocardial infarction.

[0073] Establishment of the hypoxia / reoxygenation (H / R) injury model: 2nd-3rd generation cardiomyocytes were obtained, digested, and then injected with 1×10⁻⁶ cells. 6 The culture plates were seeded at a density of 1 / 2 well and cultured for 24 h. Then, they were placed in a three-gas incubator (94% N2, 5% CO2, and 1% O2) at 37°C for 6 h of hypoxia. The plates were then removed from the hypoxia incubator, washed three times with PBS, and replaced with DMEM complete medium containing 20% ​​FBS (i.e., reoxygenation solution). The plates were then returned to a normal incubator (95% air, 5% CO2) at 37°C for 4 h of reoxygenation to establish a hypoxia-reoxygenation injury model.

[0074] The CCK-8 assay showed that after administration of compound VII, compared with the hypoxia / reoxygenation model (H / R group), the 20 μM / L concentration significantly improved cell viability. Figure 3 ).

[0075] 2. Cells were seeded in 24-well plates, with 1 ml of DMEM medium containing 10% fetal bovine serum added to each well. The plates were incubated at 37°C and 5% CO2 for 24 hours. After cell confluence, the cells were washed with PBS, followed by incubation with WSP-1 (5.00 μM) and Hoechst dye (10.0 μg / mL) for 5 minutes. The cells were washed again with PBS and cultured in FBS-free DMEM medium. One group received compound VII (20.0 μM), while the control group did not. Incubation lasted 30 minutes. Before imaging, the cells were washed with PBS and infiltrated with 2 mL of PBS.

[0076] Compound VII incubation group showed a clear green fluorescence with the hydrogen sulfide probe WSP-1, while the control group showed no fluorescence reaction, demonstrating that the compound can release hydrogen sulfide into cells and produce a corresponding therapeutic effect. Figure 4 ).

[0077] Example 4 This invention also explores the connection of compounds having the structure of Formula I (Compound I) with aromatic amines, aliphatic amines, or aliphatic alcohols. When Compound I is connected to aromatic amines, aliphatic amines, or aliphatic alcohols, the reaction does not require a catalyst, condensing agent, or strongly basic and drastic conditions. Specifically: Method A: Taking the synthesis of 4-thiocarbamoylphenyl N-phenylethylcarbamate (Va) as an example, the specific steps are as follows: In a clean, dry 50 mL round-bottom flask, compound I (1.46 mmol) was weighed and 10 mL of ultra-dry DMF was added. Phenethylamine (1.46 mmol) was added dropwise, and the mixture was stirred continuously at room temperature for 17 h. After the reaction was completed, the mixture was extracted with a large amount of water and ethyl acetate. The organic phase was dried with anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The crude product was purified by rapid column chromatography under medium pressure (dichloromethane and methanol as the developing solvent, 1% methanol) to obtain 318 mg of yellow solid, which is the product 4-thiocarbamoylphenyl N-phenylethylcarbamate (Va), with a yield of 72.7% and a temperature range of 137.0-138.5.0℃. 1 H NMR (400 MHz, DMSO) δ 9.88(s, 1H), 9.51 (s, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.95 – 7.87 (m, 2H), 7.35 –7.19 (m, 5H), 7.14 – 7.08 (m, 2H), 3.31 (dt, J = 7.9, 6.2 Hz, 2H), 2.80 (dd,J = 8.2, 6.6 Hz, 2H); 13 C NMR (101 MHz, DMSO) δ 199.51, 154.27, 153.87, 139.55,136.48, 129.20, 129.12, 128.84, 126.67, 121.40, 42.51, 35.66; HR-MS, m / z, C 16 H 16 N₂O₂S [M+H] + Theoretical value: 301.1005; measured value: 301.1011.

[0078] Method B: Same as Method A, except that stirring continuously at room temperature for 17 h is replaced with stirring continuously at 60°C for 17 h.

[0079] Method C: An excess of alcohol (248 mmol) was added to compound I (1.46 mmol), and the mixture was stirred continuously at room temperature for 17 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the crude product was purified by rapid column chromatography under medium pressure.

[0080] Method D: Same as Method C, except that after the two raw materials are added, the continuous stirring at room temperature for 17 hours is replaced with continuous stirring at 60°C for 17 hours.

[0081] The following compounds were synthesized using the methods described above: 4-Thiocarbamoylphenyl N-methyl-N-phenylcarbamate (Vb) Method A: 260 mg yellow solid, yield 62.36%, 167.5-169.0℃; 1 H NMR (400 MHz, DMSO) δ 9.89 (s, 1H), 9.52 (s, 1H), 7.92 (s, 2H), 7.60 – 7.00 (m, 7H), 3.35 (s, 3H); 13 C NMR (101 MHz, DMSO) δ 199.43, 153.83, 153.19, 143.11, 136.94,129.50, 129.14, 127.03, 126.47, 121.67, 38.50; HR-MS, m / z, C 15 H 14 N₂O₂S [M+H] + Theoretical value: 287.0849; measured value: 287.0798.

[0082] 4-Thiocarbamoylphenyl N-butylcarbamate (Vc) Method A: 63 mg pale yellow solid, yield 17.17%, 144.5-145.5℃; 1 H NMR (400 MHz, DMSO) δ 9.87 (s, 1H), 9.50 (s, 1H), 7.97 – 7.89 (m, 2H), 7.86 (t, J = 5.7 Hz, 1H), 7.13 (d, J = 8.8 Hz, 2H), 3.06 (td, J = 7.0, 5.7 Hz, 2H), 1.50 – 1.40(m, 2H), 1.32 (ddd, J = 10.0, 7.6, 6.0 Hz, 2H), 0.89 (t, J = 7.3 Hz, 3H); 13 CNMR (101 MHz, DMSO) δ 199.53, 154.29, 153.93, 136.44, 129.11, 121.43, 31.76,19.91, 19.84, 14.14; HR-MS, m / z, C 12 H 16 N₂O₂S [M+H] + Theoretical value: 253.1005; Measured value: 253.1011.

[0083] 4-Thiocarbamoylphenyl N,N-diethylcarbamate (V d) Method A, 80 mg yellow solid, yield 21.77%, 1H NMR (400 MHz, DMSO) δ 9.88 (s, 1H), 9.51 (s, 1H), 8.02 – 7.85 (m, 2H), 7.24 – 7.08 (m, 2H), 3.45 – 3.25 (m, 4H), 1.15 (dt, J = 32.0, 7.0 Hz, 6H); 169.2–171.3 °C; 1 H NMR (400 MHz, DMSO) δ9.88 (s, 1H), 9.51 (s, 1H), 8.02 – 7.85 (m, 2H), 7.24 – 7.08 (m, 2H), 3.45 –3.25 (m, 4H), 1.15 (dt, J = 32.0, 7.0 Hz, 6H); 13 C NMR (101 MHz, DMSO) δ199.45, 154.06, 153.33, 136.60, 129.09, 121.63, 42.26, 42.02, 14.66, 13.73; HR-MS, m / z, C 12 H 16 N₂O₂S [M+H] + Theoretical value: 253.1005; Measured value: 253.1000.

[0084] 4-Thiocarbamoylphenylphenylethyl carbonate (Ve) Method B, 217 mg yellow solid, yield 49.45%, 105.0-106.5℃; 1 H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 9.57 (s, 1H), 7.93 (d, J = 8.8 Hz, 2H), 7.37 – 7.20 (m,8H), 4.44 (t, J = 6.7 Hz, 2H), 3.00 (t, J = 6.7 Hz, 2H); 13 C NMR (101 MHz, DMSO) δ 199.37, 153.15, 153.02, 137.89, 137.79, 129.42, 129.32, 128.93,127.02, 121.17, 69.53, 34.63; HR-MS, m / z, C 16 H15 NO3S [M+H] + Theoretical value: 302.0845, measured value: 302.0839.

[0085] 4-Thiocarbamoylphenyl(1-phenylethyl) carbonate (Vf) Method B: 104 mg of yellow oily liquid, yield 11.85%, 115.0-116.5℃; 1 H NMR (400 MHz, DMSO) δ 9.94 (s, 1H), 9.55 (s, 1H), 8.06 – 7.89 (m, 2H), 7.53 – 7.16 (m, 7H), 5.80 (d, J = 6.6 Hz, 1H), 1.60 (d, J = 6.6 Hz, 3H); 13 C NMR (101 MHz, DMSO) δ199.33, 153.13, 152.45, 140.93, 137.77, 129.33, 129.08, 128.76, 126.49,121.17, 77.74, 22.45; HR-MS, m / z, C 16 H 15 NO3S [M+H] + Theoretical value: 302.0845, measured value: 302.0844.

[0086] Benzyl 4-thiocarbamoylphenyl carbonate (V g) Method B, 417 mg of orange-yellow solid, yield 49.83%, 114.5-116.0℃; 1 H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 9.57 (s, 1H), 7.95 (d, J = 8.7 Hz, 2H), 7.48 – 7.39 (m,5H), 7.34 – 7.28 (m, 2H), 5.29 (s, 2H); 13 C NMR (101 MHz, DMSO) δ 199.37,153.20, 153.06, 137.86, 137.77, 135.35, 129.36, 129.11, 129.07, 128.91,121.20, 70.52; HR-MS, m / z, C 15 H 13 NO3S [M+H] + Theoretical value: 288.0689; measured value: 288.0704.

[0087] Cyclohexyl 4-thiocarbamoylphenyl carbonate (V h) Method B, 291 mg orange-yellow solid, yield 35.77%, 143.5-145.0℃; 1 H NMR (400 MHz, DMSO) δ 9.94 (s, 1H), 9.55 (s, 1H), 8.07 – 7.86 (m, 2H), 7.41 – 7.19 (m, 2H), 4.66 (td, J = 8.9, 4.4 Hz, 1H), 1.91 (dd, J = 12.4, 5.4 Hz, 2H), 1.77 – 1.63 (m, 2H), 1.57 – 1.44 (m, 3H), 1.42 – 1.23 (m, 3H); 13 C NMR (101 MHz, DMSO) δ199.37, 153.22, 152.50, 137.68, 129.30, 121.20, 77.89, 31.29, 25.16, 25.12,23.46; HR-MS, m / z, C 14 H 17 NO3S [M+H] + Theoretical value 280.1002, measured value 280.1011.

[0088] Methyl 4-thiocarbamoylphenyl carbonate (V i) Method C, 251 mg yellow solid, yield 81.6%, 183.5-184.5℃; 1 H NMR (400 MHz, DMSO)δ 9.94 (s, 1H), 9.56 (s, 1H), 7.99 – 7.88 (m, 2H), 7.35 – 7.23 (m, 2H), 3.84 (s, 3H); 13 C NMR (101 MHz, DMSO) δ 199.40, 153.72, 153.22, 137.84, 129.32,121.20, 56.10; HR-MS, m / z, C9H9NO3S [M+H] + Theoretical value: 212.0376; Measured value: 212.0372.

[0089] Isopropyl 4-thiocarbamoylphenyl carbonate (Vj) Method D, 105 mg yellow solid, yield 30.13%, 170-171.5℃;1 H NMR (400 MHz, DMSO)δ 9.94 (s, 1H), 9.56 (s, 1H), 8.07 – 7.79 (m, 2H), 7.37 – 7.18 (m, 2H), 4.88(p, J = 6.2 Hz, 1H), 1.31 (d, J = 6.2 Hz, 6H); 13 C NMR (101 MHz, DMSO) δ199.38, 153.21, 152.56, 137.68, 129.30, 121.22, 73.65, 21.86; HR-MS, m / z, C 11 H 13 NO3S [M+H] + Theoretical value: 240.0689; Measured value: 240.0697.

[0090] 2,5,8,11,14-pentahexadecyl 4-thiocarbamoylphenyl carbonate (Vk) Method B, yellow oil, 416 mg, yield 33.1%. 1 H NMR (400 MHz, DMSO) δ 9.94 (s,1H), 9.55 (s, 1H), 7.93 (d, J = 8.8 Hz, 2H), 7.29 (d, J = 8.8 Hz, 2H), 4.36 –4.30 (m, 2H), 3.72 – 3.66 (m, 2H), 3.58 – 3.47 (m, 16H), 3.22 (s, 3H); 13 C NMR(101 MHz, DMSO) δ 199.36, 153.18, 153.15, 137.79, 129.32, 121.16, 71.73,70.27, 70.25, 70.04, 68.45, 68.42, 58.51; HR-MS, m / z, C 19 H 29 NO8S [M+H] + Theoretical value: 432.1687; Measured value: 432.1361.

[0091] 1,1,1,3,3,3-Hexafluoroprop-2-yl-4-thiocarbamoylphenyl carbonate (V l) Method C, 251 mg pale yellow solid, yield 54.0%, 121-122℃; 1H NMR (400 MHz, DMSO) δ10.01 (s, 1H), 9.62 (s, 1H), 8.06 – 7.92 (m, 2H), 7.51 – 7.39 (m, 2H), 6.98 (p, J = 6.1 Hz, 1H); 13 C NMR (101 MHz, DMSO) δ 199.21, 152.46, 150.90, 138.84,129.57, 120.87, 70.40; HR-MS, m / z, C 11 H7F6NO3S [M+H] + Theoretical value: 348.0124, measured value: 348.0101.

[0092] .

[0093] The reaction of compound I with primary amines, secondary amines and alcohols with different substitutions was investigated. The target compound (Vb ~ Vl) could be successfully isolated in good yield. The product obtained by reacting with phenol was too reactive and could not exist stably.

[0094] Of the four carbamate derivatives generated, the yields of amines containing benzene ring substitution were significantly higher than those of aliphatic amines (Va, Vb > Vc, Vd), demonstrating that the presence of a benzene ring favors the reaction. The yields of electron-rich aromatic ring-substituted amines were higher than those of electron-deficient aromatic ring-substituted amines (Va > Vb), indicating that the higher electron cloud density of the aromatic ring substitution environment better suits the electron requirements of the reaction.

[0095] Of the eight ester derivatives generated, alcohols containing benzene ring substitutions showed higher yields, and the smaller the steric hindrance, the higher the yield (Ve, Vg > Vf), demonstrating that the conjugation effect and electron delocalization of the aromatic ring favor the reaction, while larger steric hindrance is detrimental. Shorter aliphatic alkyl chains and fewer branches resulted in higher yields, indicating that smaller steric hindrance favors the reaction (Vi > Vk, Vi > Vj > Vh). Alcohols substituted with strong electron-withdrawing groups had higher yields than aliphatic alcohols other than methanol (Vl > Vh, Vl, Vj, Vk), suggesting that strong electron-withdrawing groups may stabilize the electrophilic center, thus favoring the reaction.

[0096] Determination of hydrogen sulfide release: (1) Plotting the sodium sulfide calibration curve Prepare a 5 mM Na₂S sodium phosphate buffer solution (100 mM, pH 7.2-7.4) as a stock solution. (Preparation method: Dissolve 39.25 mg of Na₂S, M=78.04, in a 100 mL volumetric flask). Take 5, 10, 20, 40, 60, 80, 100, and 150 μL of Na₂S stock solution and add them to 5 mL volumetric flasks. Dissolve in sodium phosphate buffer to obtain standard solutions of 5, 10, 20, 40, 60, 80, 100, and 150 μM, respectively. Transfer each solution to a 15 mL brown light-proof EP tube. Then, add 2.5 mL of methylene blue (MB+) mixture to each group for reaction: 30 mM FeCl₃ (200 μL) dissolved in 1.2 M HCl, 20 mM N,N-dimethyl-p-phenylenediamine C₈H₂O. 12 N2 (200 μL) was dissolved in 7.2 M HCl, and 1% (w / v) zinc acetate Zn(CH3COO)2 (100 μL) was dissolved in deionized water. The reaction was carried out in the dark at room temperature for 20 minutes (each reaction was performed in triplicate). The absorbance of methylene blue in 150 μL of the mixture was measured at λmax = 670 nm using a 96-well microplate reader (BioTek). A Na2S calibration curve was plotted. Figure 5 ).

[0097] y = 0.0046x + 0.0258; R 2 = 0.9961 (2) Determination of hydrogen sulfide release from the compound Prepare a 10 mM L-cysteine ​​sodium phosphate buffer (100 mM, pH 7.2-7.4).

[0098] (Preparation method: L-cysteine, M=121.16, 12 mg dissolved in a 10 mL volumetric flask).

[0099] Prepare a 10 mM DMSO solution of the compound as a stock solution, taking compound Va as an example: (Preparation method: Compound Va, M=300.38, 15 mg dissolved in a 5 mL volumetric flask). Prepare methylene blue mixture in advance; then take 50 μL of the compound stock solution and add it to 3.33 mL of sodium phosphate buffer (100 mM, pH 7.2-7.4), then add 1.66 mL of methylene blue mixture, add 50 μL of L-cysteine, shake well to obtain 100 μM compound working solution (total 5 mL). Measure the absorbance of methylene blue in 150 μL of the mixture at λmax = 670 nm using a 96-well microplate reader (BioTek) every 20 minutes. Substitute the absorbance into the calibration curve to obtain the hydrogen sulfide release of each compound. Figure 6 ).

[0100] Depend on Figure 6It can be seen that compound Vb releases the most significantly. The curves of amine-substituted derivatives show similar trends, all increasing rapidly from 0 to 20 min and then stabilizing, except for Vd and compound II, which decrease from 160 to 180 min, but the hydrogen sulfide concentration ranges are different; the hydrogen sulfide concentrations of alcohol-substituted derivatives are basically concentrated in the 15-25 μM range, with similar fluctuation trends.

[0101] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A TBZ-type hydrogen sulfide donor derivative, characterized in that, It has the structure shown in Equation I: Formula I.

2. The method for preparing the TBZ-type hydrogen sulfide donor derivative as described in claim 1, characterized in that, Includes the following steps: In organic solvent 1, 1,1'-carbonyldiimidazole and 4-hydroxythiobenzamide were mixed and reacted to obtain 4-thiocarbamoylbenzene 1H-imidazol-1-carboxylate. In organic solvent 2, the 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate and methanesulfonic acid are mixed and reacted to obtain the TBZ-type hydrogen sulfide donor derivative.

3. The preparation method according to claim 2, characterized in that, The molar ratio of 1,1'-carbonyldiimidazole to 4-hydroxythiobenzamide is (50~52):(25~26); the reaction temperature is room temperature, the reaction is monitored by thin-layer chromatography, and the reaction ends when 4-hydroxythiobenzamide disappears.

4. The preparation method according to claim 2, characterized in that, The molar ratio of 4-thiocarbamoylbenzene 1H-imidazolium-1-carboxylate to methanesulfonic acid is 1:(0.9~1.1); the reaction 2 is carried out at room temperature for 2~3 hours with stirring.

5. A method for preparing an H2S donor conjugate, characterized in that, When the drug is mexiletine hydrochloride, the preparation method includes the following steps: In organic solvent 3, mexiletine hydrochloride was mixed with a compound with the structure shown in Formula I and reacted to obtain product compound II. The structural formula of compound II is: ; When the drug is propranolol hydrochloride, the preparation method includes the following steps: (1) In an alkaline aqueous system, propranolol hydrochloride and ditert-butyl dicarbonate were reacted to obtain compound III; (2) In organic solvent 4, N-fluorenylmethoxycarbonyl-β-alanine, 4-dimethylaminopyridine, compound III, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride were reacted to obtain compound IV; (3) In organic solvent 5, compound IV and piperidine are reacted to obtain compound V; (4) In organic solvent 6, the compound shown in formula I and compound V are reacted to obtain compound VI; (5) After mixing compound VI and TFA in DCM solution, reaction 8 was carried out to obtain target compound VII; The structural formula of compound III is: ; The structural formula of compound IV is: ; The structural formula of compound V is: ; The structural formula of compound VI is: ; The structural formula of compound VII is: .

6. The preparation method according to claim 5, characterized in that, The molar ratio of mexiletine hydrochloride to the compound with the structure shown in Formula I is 1:(0.9~1.1); the reaction 3 is carried out at room temperature for 17~19 h.

7. The preparation method according to claim 5, characterized in that, In step (1), the molar ratio of propranolol hydrochloride and ditert-butyl dicarbonate is 1:(1.1~1.3), the reaction temperature is room temperature, and the reaction is monitored by TLC.

8. The preparation method according to claim 5, characterized in that, In step (2), the molar ratio of N-fluorenemethoxycarbonyl-β-alanine, 4-dimethylaminopyridine, compound III, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride is (1.0~1.2):(4~5):1.0:(2~3); the reaction temperature is room temperature, and the reaction is monitored by TLC.

9. The preparation method according to claim 5, characterized in that, In step (3), the molar ratio of compound IV to piperidine is 1:(4~6), the reaction temperature is room temperature, and the reaction is monitored by TLC.

10. The preparation method according to claim 5, characterized in that, In step (4), the molar ratio of compound I to compound V is (1.5~2):(2~2.5), the temperature of reaction 7 is 50℃, and the time is 4h; In step (5), the molar ratio of compound VI to TFA is 1:(5-10); the reaction 8 is carried out under ice bath conditions and the reaction is monitored by TLC.

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