Isothiocyanate derivative, preparation method and application thereof

Abstract

The present invention discloses an isothiocyanate derivative, its preparation method, and application. The structural formula of the isothiocyanate derivative of the present invention is shown in formula (I): #imgabs0# wherein X is a sulfoxide or sulfone group, R is -S-R1 or -NH-R2, and R1 and R2 are each independently a saturated alkyl group, an aromatic hydrocarbon group, or a heterocycle. By structurally modifying and optimizing the isothiocyanate, not only can the formation of unstable compounds caused by the thiocyanate structure's susceptibility to reaction with the protic solvent water in the body be reduced, but the level of inflammatory factors and oxidative stress damage can also be reduced. The isothiocyanate derivative has the characteristics of high efficiency, low toxicity, and structural stability, and can be widely used in the preparation of drugs for treating ischemic cardiomyopathy or myocardial ischemia-reperfusion injury.

Description

Technical Field

[0001] The present invention relates to the technical field of organic synthesis, and more particularly to an isothiocyanate derivative and a preparation method and application thereof. Background Art

[0002] In recent years, with the development of coronary intervention techniques and antithrombotic drugs, the mortality rate of ischemic cardiovascular disease has greatly increased. However, myocardial ischemia-reperfusion injury (MIRI) after myocardial infarction leads to post-MI ventricular remodeling, seriously affecting patients' quality of life and even causing death. MIRI is primarily caused by a variety of factors that reduce coronary blood flow, leading to obstructed myocardial blood supply, insufficient nutrient supply, and reduced clearance of metabolic products, resulting in myocardial cell damage and even death.

[0003] Numerous studies have shown that the occurrence of MIRI may be related to mechanisms such as calcium overload, increased oxygen free radicals, impaired myocardial fiber energy metabolism, inflammation, and acidosis. It is a complex pathophysiological process involving a series of mechanisms, including inflammation, oxidative stress, apoptosis, and autophagy. Furthermore, myocardial tissue includes cardiomyocytes, endothelial cells, vascular smooth muscle cells, fibroblasts, and macrophages, which account for approximately 75% of the myocardial volume. Studies have shown that cardiac microvascular endothelial cells far outnumber cardiomyocytes and can directly affect cardiomyocytes by secreting active substances, causing myocardial hypoxic injury.

[0004] Isothiocyanates (ITCs), enzymatic degradation products of glucosinolates (glucosinolates), are widely and abundantly present in Brassica vegetables of the cruciferous family. In recent years, in-depth research has revealed that these ITCs also possess pharmacological effects such as oxidative stress reduction, anti-inflammatory and antibacterial properties, and immune enhancement. However, the thiocyanate structure readily reacts with the protic solvent water in the body to form unstable compounds, limiting their further clinical application. Summary of the Invention

[0005] The present invention aims to overcome the defects and shortcomings of existing isothiocyanates, that is, the thiocyanate structure easily reacts with the protic solvent water in the body to form unstable compounds, and to provide an isothiocyanate derivative.

[0006] Another object of the present invention is to provide a method for preparing the above-mentioned isothiocyanate derivatives.

[0007] Another object of the present invention is to provide the use of the above-mentioned isothiocyanate derivatives in the preparation of drugs for resisting myocardial hypoxia-reoxygenation injury.

[0008] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0009] An isothiocyanate derivative, the structural formula of which is shown in formula (I):

[0010]

[0011] Wherein, X is a sulfoxide group or a sulfone group, R is -S-R1 or -NH-R2, and R1 and R2 are each independently a saturated alkyl group, an aromatic hydrocarbon group or a heterocycle.

[0012] Preferably, the saturated alkyl group is any one of ethyl, propyl and isobutyl.

[0013] Preferably, the aromatic hydrocarbon group is any one of p-methylphenyl, m-methylphenyl, p-ethylphenyl, p-methoxyphenyl, p-bromophenyl and p-bromobenzyl.

[0014] Preferably, when R is -NH-R2, R2 is an aromatic hydrocarbon group, and the aromatic hydrocarbon group is p-methylphenyl or p-bromophenyl.

[0015] Preferably, the heterocycle is 2-methyl-3-furyl or 2-furylmethylene.

[0016] A method for preparing the above-mentioned isothiocyanate derivative comprises the following steps:

[0017] After fully reacting the reaction monomer I with the reaction monomer II at room temperature, the isothiocyanate derivative can be obtained through extraction, separation and purification;

[0018] Wherein, the monomer I is or

[0019]

[0020] The monomer II is HS-R1 or H2N-R2, and R1 and R2 are each independently a saturated alkyl group, an aromatic hydrocarbon group or a heterocycle.

[0021] Specifically, monomer Ia can be prepared by the following preparation method:

[0022] In an ice-salt bath, the raw materials (recorded as formula II, 1.3300g,

[0023] 9.0323mmol) was dissolved in 30mL of dichloromethane and transferred to a 250mL round-bottom flask. 85% MCPBA (1.5618g, 9.0502mmol) dissolved in dichloromethane was slowly added dropwise to the round-bottom flask containing the raw material formula II. The reaction temperature was maintained for more than 5min and the solution reaction temperature was always below 0°C. The reaction progress was monitored by TLC (developing solvent V 二氯甲烷 :V 甲醇=20:1), stirred and reacted for 1 hour, extracted with saturated NaHCO3 solution, combined the organic layers, and distilled under reduced pressure to obtain a residue, which was separated and purified by column chromatography with gradient elution (eluent V 石油醚 :V 乙酸乙酯 =20:1~1:1), and drying to obtain pure monomer Ia.

[0024] Monomer Ib is obtained by the following preparation method:

[0025] The raw materials (denoted as Formula II) is dissolved in an organic solvent and transferred to a reaction apparatus, and then an oxidant is added. The mixture is stirred and reacted for more than 15 minutes at room temperature for 3.5 hours, and then extracted with an alkaline solution. The monomer Ib is separated, purified, and dried. The molar ratio of the raw material Formula II to the oxidant is 1:4.

[0026] Preferably, the molar ratio of monomer I to monomer II is 1:(1-1.5).

[0027] Preferably, when the monomer II is HS-R1, the isothiocyanate derivative can be obtained by reacting the monomer I and the monomer II in an alkaline solution at room temperature for 0.5 to 10 hours.

[0028] Specifically, under the action of an alkaline solution, monomer I and HS-R1 (monomer II) are dissolved in an organic solvent and mixed in a round-bottom flask, stirred and reacted at room temperature for 0.5 to 10 hours, filtered, washed, and distilled under reduced pressure to remove the organic solvent, the residue is extracted with an alkaline solution, and the solvent is again distilled under reduced pressure. The residue is purified and separated by column chromatography to obtain an isothiocyanate derivative; wherein the molar ratio of monomer I to the alkaline solution is 1:1.05; and the molar ratio of monomer I to HS-R1 (monomer II) is 1:(1 to 1.5);

[0029] Preferably, when the monomer II is H2N-R2, the isothiocyanate derivative can be obtained after the monomer I and monomer II react at room temperature for 24 hours;

[0030] The molar ratio of the monomer I to the monomer II is 1:(1.1-1.5).

[0031] Specifically, at room temperature, monomer I is dissolved in an organic solvent (acetonitrile solution) and transferred to a round-bottom flask. H2N-R2 (monomer II) is added under stirring. After stirring and reacting for 24 hours, the mixture is filtered and the organic solvent is removed by distillation under reduced pressure to obtain a residue. The residue is separated and purified by column chromatography and dried to obtain an isothiocyanate derivative.

[0032] The use of the above-mentioned isothiocyanate derivative in the preparation of a drug for resisting myocardial hypoxia-reoxygenation injury is also within the scope of protection of the present invention.

[0033] Compared with the prior art, the present invention has the following beneficial technical effects:

[0034] The present invention provides an isothiocyanate derivative. By structurally modifying and optimizing the isothiocyanate, not only can the generation of unstable compounds due to the susceptibility of the thiocyanate structure to react with the protic solvent water in the body be reduced, but the level of inflammatory factors and oxidative stress damage can also be reduced. The isothiocyanate derivative has the characteristics of high efficiency, low toxicity and structural stability, and can be widely used in the preparation of drugs for treating ischemic cardiomyopathy or myocardial ischemia-reperfusion injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The graph shows the stability of 3-(methylsulfinyl)propyl isothiocyanate and 3-(methylsulfonyl)propyl isothiocyanate derivatives of the present invention in protic solvents and aprotic solvents.

[0036] Figure 2 3-(methylsulfinyl)propyl isothiocyanate and 3-(methylsulfonyl)propyl isothiocyanate derivatives of the present invention at different concentrations of cell viability graph.

[0037] Figure 3 This is the effect of compound 2-3, compound 2-9, compound 3-3, and compound 3-9 of the present invention on the reactive oxygen species in CMECs cells. DETAILED DESCRIPTION

[0038] The present invention will be further described below in conjunction with specific embodiments, but the examples do not limit the present invention in any form. Unless otherwise specified, the raw materials and reagents used in the examples of the present invention are conventionally purchased raw materials and reagents.

[0039] Monomer Ia

[0040] The structural formula of monomer Ia is: It can be prepared by the following preparation method:

[0041] In an ice-salt bath, the raw materials (denoted as formula II, 1.3300 g, 9.0323 mmol) was dissolved in 30 mL of dichloromethane and transferred to a 250 mL round-bottom flask. 85% MCPBA (1.5618 g, 9.0502 mmol) dissolved in dichloromethane was slowly added dropwise to the round-bottom flask containing the raw material formula II. The reaction temperature was maintained for more than 5 minutes and the solution reaction temperature was always below 0°C. The reaction progress was monitored by TLC (developing solvent V 二氯甲烷 :V 甲醇 =20:1), stirred and reacted for 1 hour, extracted with saturated NaHCO3 solution, combined the organic layers, and distilled under reduced pressure to obtain a residue, which was separated and purified by column chromatography with gradient elution (eluent V石油醚 :V 乙酸乙酯 =20:1~1:1), and drying to obtain pure monomer Ia.

[0042] Monomer Ib

[0043] The structural formula of monomer Ib is: It can be prepared by the following preparation method:

[0044] At room temperature, the raw materials The product (denoted as Formula II, 1.3349 g, 9.0655 mmol) was dissolved in 30 mL of dichloromethane and transferred to a 250 mL round-bottom flask. 85% MCPBA (6.2125 g, 36.067 mmol) dissolved in dichloromethane was slowly added dropwise to the round-bottom flask containing the raw material Formula II. The reaction was maintained for more than 15 min. The reaction progress was monitored by TLC (developing agent DCM). After stirring and reacting for 3.5 h, the product was extracted with a saturated NaHCO solution. The organic layers were combined and distilled under reduced pressure to obtain a residue. The obtained residue was separated and purified by column chromatography using a gradient elution method (eluent V 石油醚 :V 乙酸乙酯 =10:1~1:1), and drying to obtain pure monomer Ib.

[0045] Example 1

[0046] A 3-(methylsulfinyl)propyl isothiocyanate derivative is [3-(methylsulfinyl)propyl]aminodithiocarbamate (denoted as compound 2-1), and its structural formula is as follows:

[0047]

[0048] The above compound 2-1 can be prepared by the following preparation method:

[0049] At room temperature, alkali metal salt NaHCO3 was dissolved in a 50 mL round-bottom flask containing 5 mL of water, and 10 mL of anhydrous ethanol was added. Monomer I-a (0.1634 g, 1.002 mmol) and ethanethiol (0.0641 g, 1.03 mmol) were added to the round-bottom flask in sequence under stirring. The reaction progress was monitored by TLC (developer V 乙酸乙酯 :V 甲醇 =10:1), react for 3 hours, filter, remove ethanol by distillation under reduced pressure, extract the aqueous layer with dichloromethane, take the organic layer, distill under reduced pressure to obtain a residue, and the obtained residue is separated and purified by column chromatography with gradient elution (eluent V 石油醚 :V 乙酸乙酯 =95:5~5:95), and dried to obtain pure compound 2-1 as a white solid with a melting point of 88.3~88.6°C and a yield of 100.00%.

[0050] Compound 2-1 was subjected to nuclear magnetic resonance (NMR) detection using a Bruker Avance NEO 600 NMR spectrometer. The results are as follows:

[0051] 1 H NMR (600 MHz, DMSO): δ = 9.95 (s, 1H), 3.69 (dd, J = 12.5, 7.0 Hz, 2H), 3.15 (q, J = 7.3 Hz, 2H), 2.82–2.65 (m, 2H), 2.53 (s, 3H), 1.94 (td, J = 13.6, 7.0 Hz,2H),1.23(t,J=7.3 Hz,3H);

[0052] 13 C NMR (600MHz, DMSO): δ = 196.96, 51.00, 45.91, 38.44, 28.73, 21.45, 14.91.

[0053] MS (ESI) (m / z): 225; (M+H + ):226.

[0054] Example 2

[0055] A 3-(methylsulfinyl)propyl isothiocyanate derivative is 4-bromophenyl-[3-(methylsulfinyl)propyl]carbamyldithioate (denoted as compound 2-2), and its structural formula is as follows:

[0056]

[0057] The above compound 2-2 can be prepared by the following preparation method:

[0058] At room temperature, alkali metal salt NaHCO3 was dissolved in a 50 mL round-bottom flask containing 5 mL of water, and 10 mL of anhydrous ethanol was added. Monomer Ia (0.1645 g, 1.009 mmol) and 4-bromobenzenethiol (0.1892 g, 1.001 mol) were added to the round-bottom flask in sequence under stirring. The reaction progress was monitored by TLC (developer V 甲醇 :V 乙酸乙酯 =1:10), react for 24 hours, filter, remove ethanol by distillation under reduced pressure, extract the aqueous layer with dichloromethane, take the organic layer, distill under reduced pressure to obtain a residue, and the obtained residue is separated and purified by column chromatography with gradient elution (eluent V 甲醇 :V 乙酸乙酯 =0:1~1:20), and dried to obtain pure compound 2-2 as a white solid with a melting point of 113.4~113.6°C and a yield of 42.25%.

[0059] Compound 2-2 was subjected to nuclear magnetic resonance (NMR) detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0060] 1 H NMR (400MHz, DMSO): δ = 9.77 (s, 1H), 7.68 (d, J = 8.4Hz, 2H), 7.44 (d, J = 8.4Hz, 2H), 3.67(dd,J=12.0,6.3Hz,2H),2.82–2.62(m,2H),2.53(s,3H),1.99–1.88(m,2H);

[0061] 13 C NMR (101MHz, DMSO): δ = 194.59, 138.45, 132.90, 129.45, 124.65, 50.89, 46.15, 38.43, 21.37.

[0062] MS (ESI) (m / z): 351; (M+H + ):352.

[0063] Example 3

[0064] A 3-(methylsulfinyl)propyl isothiocyanate derivative is p-tolyl-[3-(methylsulfinyl)propyl]carbamyldithioate (denoted as compound 2-3), and its structural formula is as follows:

[0065]

[0066] The above compound 2-3 can be prepared by the following preparation method:

[0067] At room temperature, alkali metal salt NaHCO3 was dissolved in a 100 mL round-bottom flask containing 5 mL of water, and 10 mL of anhydrous ethanol was added. Monomer I-a (0.1688 g, 1.036 mmol) and 4-methylthiophenol (0.1262 g, 1.016 mmol) were added to the round-bottom flask in sequence under stirring. The reaction progress was monitored by TLC (developer V 乙酸乙酯 :V 甲醇 =5:1), react for 4 hours, filter, remove ethanol by distillation under reduced pressure, extract the aqueous layer with dichloromethane, take the organic layer, distill under reduced pressure to obtain a residue, and the obtained residue is separated and purified by column chromatography with gradient elution (eluent V 二氯甲烷 :V 甲醇 =1:0~20:1), and dried to obtain pure compound 2-3 as a white solid with a melting point of 113.0~113.6°C and a yield of 68.22%.

[0068] Compound 2-3 was detected by nuclear magnetic resonance using a Bruker Avance NEO 400 nuclear magnetic resonance spectrometer. The results are as follows:

[0069] 1 H NMR (400MHz, DMSO): δ = 9.50 (s, 1H), 7.39 (d, J = 8.0Hz, 2H), 7.30 (d, J = 8.0Hz, 2H), 3.67 ( dd,J=12.5,6.4Hz,2H),2.82–2.60(m,2H),2.53(s,3H),2.37(s,3H),1.99–1.86(m,2H);

[0070] 13 C NMR (101MHz, DMSO): δ=191.96,137.19,130.88,126.51,120.29,46.63,39.96,33.97,17.28,16.75.

[0071] MS (ESI) (m / z): 287; (M+H + ):288.

[0072] Example 4

[0073] A 3-(methylsulfinyl)propyl isothiocyanate derivative is 1-(4-bromophenyl)-3-[3-(methylsulfinyl)propyl]thiourea (denoted as compound 2-4), and its structural formula is as follows:

[0074]

[0075] The above compound 2-4 can be prepared by the following preparation method:

[0076] At room temperature, monomer Ia (0.6500 g, 3.988 mmol) was dissolved in 30 mL of acetonitrile and transferred to a 100 mL round-bottom flask. 4-bromoaniline (0.7588 g, 4.411 mmol) was added under stirring. The reaction progress was monitored by TLC (developing solvent V 二氯甲烷 :V 甲醇 =20:1), stirred and reacted for 24 hours, filtered, and the organic solvent was removed by distillation under reduced pressure to obtain a residue, which was then separated and purified by column chromatography using a gradient elution (eluent V 二氯甲烷 :V 甲醇 =1:0~20:1), and dried to obtain pure compound 2-4 as a white solid with a melting point of 149.3~149.7°C and a yield of 91.28%.

[0077] Compounds 2-4 were subjected to NMR detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0078] 1 H NMR (400MHz, DMSO): δ = 9.60 (s, 1H), 7.95 (s, 1H), 7.50 (d, J = 11.7Hz, 2H), 7.39 (d, J = 8 .8Hz,2H),3.59(d,J=5.5Hz,2H),2.82–2.66(m,2H),2.55(s,3H),1.96–1.88(m,2H);

[0079] 13 C NMR (101MHz, DMSO): δ = 180.94, 139.13, 131.82, 125.53, 116.53, 51.12, 43.34, 38.50, 22.36.

[0080] MS (ESI) (m / z): 334; (M+H + ):335.

[0081] Example 5

[0082] A 3-(methylsulfinyl)propyl isothiocyanate derivative is 1-[3-(methylsulfoxy)propyl-3-(p-tolyl)]thiourea (denoted as compound 2-5), and its structural formula is as follows:

[0083]

[0084] The above-mentioned isothiocyanate derivatives can be prepared by the following preparation method:

[0085] At room temperature, monomer Ia (0.6600 g, 4.049 mmol) was dissolved in 30 mL of acetonitrile and transferred to a 100 mL round-bottom flask. p-Toluidine (0.4766 g, 4.448 mmol) was added under stirring. The reaction progress was monitored by TLC (developing solvent V 二氯甲烷 :V 甲醇 =20:1), stirred and reacted for 24 hours, filtered, and the organic solvent was removed by distillation under reduced pressure to obtain a residue, which was then separated and purified by column chromatography using a gradient elution (eluent V 石油醚 :V 乙酸乙酯 =1:1~0:1), and dried to obtain pure compound 2-5 as a white solid with a melting point of 122.5~124.3°C and a yield of 96.33%.

[0086] Compounds 2-5 were subjected to NMR detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0087] 1 H NMR (400MHz, DMSO): δ = 9.43 (s, 1H), 7.72 (s, 1H), 7.24 (d, J = 8.3Hz, 2H), 7.14 (d, J = 8.3Hz, 2H), 3 .59(d,J=5.9Hz,2H),2.85–2.62(m,2H),2.54(s,3H),2.28(s,3H),1.92(dq,J=10.2,6.9Hz,2H);

[0088] 13 C NMR (101MHz, DMSO): δ = 180.94, 136.77, 134.13, 129.66, 124.24, 51.18, 43.37, 38.48, 22.48, 20.98.

[0089] MS (ESI) (m / z): 270; (M+H + ):271.

[0090] Example 6

[0091] A 3-(methylsulfinyl)propyl isothiocyanate derivative is 4-methoxyphenyl-[3-(methylsulfoxy)propyl]carbamyldithioate (denoted as compound 2-6), and its structural formula is as follows:

[0092]

[0093] The preparation method of the above-mentioned isothiocyanate derivatives 2-6 is basically the same as that of Example 3, except that the raw material is p-methoxythiophenol, and the reaction progress is tracked by TLC (developing solvent V 甲醇 :V 乙酸乙酯 =1:5), the residue obtained by post-treatment was separated and purified by column chromatography under the elution conditions of dichloromethane → V 乙酸乙酯 :V 甲醇 =10:1, the product is a white solid with a melting point of 87.6-88.2°C and a yield of 88.54%.

[0094] Compounds 2-6 were subjected to NMR detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0095] 1H NMR (400MHz, DMSO): δ = 9.37 (s, 1H), 7.42 (d, J = 6.9Hz, 2H), 7.04 (d, J = 7.0Hz, 2H), 3.82 (s, 3H) ,3.66(d,J=5.9Hz,2H),2.70(ddd,J=43.9,13.1,6.2Hz,2H),2.52(s,3H),1.96–1.86(m,2H);

[0096] 13 C NMR (101MHz, DMSO): δ = 196.34, 161.38, 138.22, 120.42, 115.62, 55.83, 50.94, 45.88, 38.44, 21.45.

[0097] MS (ESI) (m / z): 303; (M+H + ):304.

[0098] Example 7

[0099] A 3-(methylsulfinyl)propyl isothiocyanate derivative is 4-bromobenzyl-[3-(methylsulfinyl)propyl]carbamyldithioate (denoted as compound 2-9), and its structural formula is as follows:

[0100]

[0101] The preparation method of the above-mentioned isothiocyanate derivative 2-9 is basically the same as that of Example 4, except that the starting material is p-bromobenzyl mercaptan, the reaction progress is tracked by TLC (developing solvent ethyl acetate), and the elution conditions for the post-treatment column chromatography separation and purification are V 石油醚 :V 乙酸乙酯 =20:1→ethyl acetate→V 乙酸乙酯 :V 甲醇 =20:1, the product is a white solid with a melting point of 96.2-97.0°C and a yield of 98.50%.

[0102] Compounds 2-9 were subjected to NMR detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0103] 1 H NMR (400MHz, DMSO): δ = 10.10 (t, J = 4.5Hz, 1H), 7.51 (d, J = 8.4Hz, 2H), 7.33 (d, J = 8.4Hz, 2H), 4 .50(s,2H),3.70(dd,J=12.3,6.9Hz,2H),2.81–2.65(m,2H),2.53(s,3H),1.99–1.92(m,2H);

[0104] 13 C NMR (101MHz, DMSO): δ = 196.11, 137.60, 131.76, 131.54, 120.70, 50.96, 46.35, 38.46, 37.75, 21.41.

[0105] MS (ESI) (m / z): 365; (M+H + ):366.

[0106] Example 8

[0107] A 3-(methylsulfonyl)propyl isothiocyanate derivative is ethyl 3-(methylsulfonyl)propylaminodithiocarbamate (denoted as compound 3-1), and its structural formula is as follows:

[0108]

[0109] The above compound 3-1 can be prepared by the following preparation method:

[0110] At room temperature, alkali metal salt NaHCO3 was dissolved in a 50 ml round-bottom flask containing 5 mL of water, and 10 mL of anhydrous ethanol was added. Monomer Ib (0.1802 g, 1.007 mmol) and ethanethiol (0.1000 g, 1.610 mmol) were added to the round-bottom flask in sequence under stirring. The reaction progress was monitored by TLC (developing solvent V 石油醚 :V 乙酸乙酯 =1:1), react for 2 hours, filter, remove ethanol by distillation under reduced pressure, extract the aqueous layer with dichloromethane, take the organic layer, distill under reduced pressure to obtain a residue, and the obtained residue is separated and purified by column chromatography with gradient elution (eluent V 石油醚 :V 乙酸乙酯 =10:1~1:1), and dried to obtain pure compound 3-1 as a yellow liquid with a yield of 84.50%.

[0111] Compound 3-1 was subjected to nuclear magnetic resonance (NMR) detection using a Bruker Avance NEO 600 NMR spectrometer. The results are as follows:

[0112] 1 H NMR (600MHz, DMSO): δ = 9.95 (s, 1H), 3.69 (dd, J = 12.3, 6.9Hz, 2H), 3.15 (dt, J = 13.3, 6.6Hz, 4H), 2.98 (s, 3H), 2.02–1.98 (m, 2H), 1.23 (t, J = 7.3Hz, 3H);

[0113] 13C NMR (151MHz, DMSO): δ = 197.22, 51.73, 45.34, 40.58, 28.76, 21.35, 14.87.

[0114] MS (ESI) (m / z): 241; (M+H + ):242.

[0115] Example 9

[0116] A 3-(methylsulfonyl)propyl isothiocyanate derivative is 4-bromophenyl-[3-(methylsulfonyl)propyl]aminodithiocarbamate (denoted as compound 3-2), and its structural formula is as follows:

[0117]

[0118] The above compound 3-2 can be prepared by the following preparation method:

[0119] At room temperature, alkali metal salt NaHCO3 was dissolved in a 100 mL round-bottom flask containing 10 mL of water, and 20 mL of anhydrous ethanol was added. Monomer Ib (0.7170 g, 4.005 mmol) and 4-bromothiophenol (0.7573 g, 4.005 mmol) were added to the round-bottom flask in sequence under stirring. The reaction progress was monitored by TLC (developing solvent V 石油醚 :V 乙酸乙酯 =1:1), react for 4 hours, filter, remove ethanol by distillation under reduced pressure, extract the aqueous layer with dichloromethane, take the organic layer, distill under reduced pressure to obtain a residue, and the obtained residue is separated and purified by column chromatography with gradient elution (eluent V 石油醚 :V 乙酸乙酯 =1:0~1:1), and dried to obtain pure compound 3-2 as a white solid with a melting point of 127.6~129.5℃ and a yield of 82.20%.

[0120] Compound 3-2 was subjected to nuclear magnetic resonance (NMR) detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0121] 1 H NMR (400MHz, DMSO): δ = 9.75 (t, J = 5.1Hz, 1H), 7.68 (d, J = 8.4Hz, 2H), 7.44 (d, J = 8.4Hz, 2H), 3.67(dd,J=12.6,6.9Hz,2H),3.16–3.10(m,2H),2.98(s,3H),1.99(dt,J=14.9,7.4Hz,2H);

[0122] 13C NMR (101MHz, DMSO): δ = 194.88, 138.44, 132.91, 129.45, 124.67, 51.64, 45.62, 40.59, 21.24.

[0123] MS (ESI) (m / z): 367; (M+H + ):368.

[0124] Example 10

[0125] A 3-(methylsulfonyl)propyl isothiocyanate derivative is p-tolyl-[3-(methylsulfonyl)propyl]aminodithiocarbamate (denoted as compound 3-3), and its structural formula is as follows:

[0126]

[0127] The preparation method of the above compound 3-3 is basically the same as that of Example 9, except that the raw material is p-bromothiophenol, and the reaction progress is tracked by TLC (developing solvent V 乙酸乙酯 :V 石油醚 =2:1), the residue obtained by post-treatment was separated and purified by column chromatography under the elution conditions of V 石油醚 :V 乙酸乙酯 =1:0~1:2, the product is a white solid, melting point 97.9~99.2℃, and the yield is 91.58%.

[0128] Compound 3-3 was subjected to nuclear magnetic resonance (NMR) detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0129] 1 H NMR (400MHz, DMSO): δ = 9.48 (t, J = 5.0Hz, 1H), 7.39 (d, J = 8.0Hz, 2H), 7.30 (d, J = 8.0Hz, 2H), 3 .67(dd,J=12.7,6.8Hz,2H),3.14–3.09(m,2H),2.98(s,3H),2.37(s,3H),2.03–1.94(m,2H);

[0130] 13 C NMR (101MHz, DMSO): δ = 196.00, 140.62, 136.36, 130.70, 126.47, 51.66, 45.42, 40.57, 21.42, 21.29.

[0131] MS (ESI) (m / z): 286; (M+H + ):304.

[0132] Example 11

[0133] A 3-(methylsulfonyl)propyl isothiocyanate derivative is 1-[3-(methylsulfonyl)propyl]-3-(p-tolyl)thiourea (denoted as compound 3-4), and its structural formula is as follows:

[0134]

[0135] The above compound 3-5 can be prepared by the following preparation method:

[0136] At room temperature, monomer Ib (0.5694 g, 3.017 mmol) was dissolved in 30 mL of acetonitrile and transferred to a 100 mL round-bottom flask. p-Toluidine (0.3621 g, 3.379 mmol) was added under stirring. The reaction progress was monitored by TLC (developing solvent V 乙酸乙酯 :V 石油醚 =2:1), stirred and reacted for 24 hours, filtered, and the organic solvent was removed by distillation under reduced pressure to obtain a residue, which was then separated and purified by column chromatography using a gradient elution (eluent V 乙酸乙酯 :V 石油醚 =1:2~1:0), and dried to obtain pure compound 3-5 as a white solid with a melting point of 88.5~89.0°C and a yield of 73.99%.

[0137] Compounds 3-5 were detected by nuclear magnetic resonance using a Bruker Avance NEO 400 nuclear magnetic resonance spectrometer. The results are as follows:

[0138] 1 H NMR (400MHz, DMSO): δ = 9.45 (s, 1H), 7.72 (s, 1H), 7.23 (d, J = 8.3Hz, 2H), 7.14 (d, J = 8.2Hz, 2H), 3.58(dd,J=12.3,6.2Hz,2H),3.16–3.10(m,2H),2.98(s,3H),2.28(s,3H),2.01–1.93(m,2H);

[0139] 13 C NMR (101MHz, DMSO): δ = 181.02, 136.70, 134.23, 129.68, 124.31, 51.85, 42.83, 40.56, 22.35, 20.97.

[0140] MS (ESI) (m / z): 286; (M+H + ):287.

[0141] Example 12

[0142] A 3-(methylsulfonyl)propyl isothiocyanate derivative is 1-(4-bromophenyl)-3-[3-(methylsulfonyl)propyl]thiourea (denoted as compound 3-5), and its structural formula is as follows:

[0143]

[0144] The preparation method of the above compound 3-4 is basically the same as that of Example 11, except that the raw material is p-bromoaniline, and the reaction progress is tracked by TLC (developing solvent V 石油醚 :V 乙酸乙酯 =1:2), the residue obtained by post-treatment was separated and purified by column chromatography under the elution conditions of V 石油醚 :V 乙酸乙酯 =4:1~0:1, the product is a white solid, melting point 149.6~150.3℃, yield 75.00%.

[0145] Compound 3-4 was subjected to NMR detection using a Bruker Avance NEO 400 NMR spectrometer. The results are as follows:

[0146] 1H NMR (400MHz, DMSO): δ = 9.61 (s, 1H), 7.94 (s, 1H), 7.53–7.47 (m, 2H), 7.42–7.37 (m, 2H) ,3.60(d,J=5.7Hz,2H),3.17–3.12(m,2H),2.99(s,3H),1.98(dt,J=14.8,5.3Hz,2H);

[0147] 13C NMR (101MHz, DMSO): δ = 181.03, 139.07, 131.85, 125.63, 116.61, 51.81, 42.79, 40.59, 22.20.

[0148] MS (ESI) (m / z): 350; (M+H + ):351.

[0149] Example 13

[0150] A 3-(methylsulfonyl)propyl isothiocyanate derivative is 4-methoxyphenyl-[3-(methylsulfonyl)propyl]carbamyldithioate (denoted as compound 3-6), and its structural formula is as follows:

[0151]

[0152] The preparation method of the above compound 3-6 is basically the same as that of Example 9, except that the starting material is p-methoxythiophenol, the reaction progress is tracked by TLC (developing solvent: dichloromethane), and the residue obtained by post-treatment is separated and purified by column chromatography under the elution conditions of V石油醚 :V 乙酸乙酯 =1:0~1:2, the product is a white solid, melting point 117.2~117.9℃, and the yield is 98.66%.

[0153] Compounds 3-6 were detected by nuclear magnetic resonance using a Bruker Avance NEO 400 nuclear magnetic resonance spectrometer. The results are as follows:

[0154] 1 H NMR (400MHz, DMSO): δ = 9.37 (t, J = 5.3Hz, 1H), 7.43 (d, J = 8.8Hz, 2H), 7.04 (d, J = 8.8Hz, 2H), 3.82 (s ,3H),3.66(dd,J=12.7,6.9Hz,2H),3.15–3.07(m,2H),2.97(s,3H),1.97(dt,J=14.9,7.5Hz,2H);

[0155] 13 C NMR (101MHz, DMSO): δ = 196.62, 161.39, 138.21, 120.40, 115.63, 55.83, 51.65, 45.35, 40.56, 21.31.

[0156] MS (ESI) (m / z): 319; (M+H + ):320.

[0157] Example 14

[0158] A 3-(methylsulfonyl)propyl isothiocyanate derivative is 4-bromobenzyl-[3-(methylsulfonyl)propyl]carbamyldithioate (denoted as compound 3-9), and its structural formula is as follows:

[0159]

[0160] The preparation method of the above compound 3-9 is basically the same as that of Example 9, except that the starting material is 4-bromobenzyl mercaptan, the reaction progress is tracked by TLC (developing solvent: dichloromethane), and the residue obtained by post-treatment is separated and purified by column chromatography under the elution conditions of V 石油醚 :V 乙酸乙酯 =10:1→dichloromethane, the product is a white solid, melting point 110.5~111.0℃, yield 93.37%.

[0161] Compounds 3-9 were detected by nuclear magnetic resonance using a Bruker Avance NEO 400 nuclear magnetic resonance spectrometer. The results are as follows:

[0162] 1H NMR (400MHz, DMSO): δ = 10.09 (s, 1H), 7.51 (d, J = 8.4Hz, 2H), 7.33 (d, J = 8.4Hz, 2H), 4.50 (s, 2H) ),3.71(dd,J=11.2,6.8Hz,2H),3.18–3.13(m,2H),2.98(s,3H),2.01(dt,J=14.9,7.4Hz,2H);

[0163] 13 C NMR (101MHz, DMSO): δ = 196.36, 137.55, 131.77, 131.54, 120.72, 51.70, 45.80, 40.61, 37.78, 21.29.

[0164] MS (ESI) (m / z): 381; (M+H + ):382.

[0165] Performance Testing

[0166] 1. Compound stability test

[0167] Compound 2, compound 2-1, compound 2-4 and compound 2-6 of the present invention, as well as compound 3, compound 3-1, compound 3-2 and compound 3-5 were selected to prepare pure samples of isothiocyanates and their derivatives at a concentration of 2.0 mg / mL. The solvents were aprotic solvent acetonitrile and protic solvent methanol: water = 1:1 (v:v), and stored in sealed sample bottles. Each sample was shaken in a constant temperature water bath at 50°C. Every 2 hours, a small amount of sample was taken out and quickly cooled in a cold water bath (-5°C). The samples were subjected to HPLC analysis using Shimadzu LCMS-2020 to determine the degradation rate. The stability curves of the above compounds in protic solvents and aprotic solvents are shown in Figure 2. Figure 1 Shown are a) degradation rates of 3-(methylsulfinyl)propyl isothiocyanate derivatives in a protic (methanol) solvent; b) degradation rates of 3-(methylsulfinyl)propyl isothiocyanate derivatives in an aprotic (acetonitrile) solvent; c) degradation rates of 3-(methylsulfonyl)propyl isothiocyanate derivatives in a protic (methanol:water=1:1) solvent; d) degradation rates of 3-(methylsulfonyl)propyl isothiocyanate derivatives in an aprotic (acetonitrile) solvent; values ​​are described as mean ± SEM (standard error of the mean, n=3).

[0168] from Figure 1a) It can be seen that as the time of the 3-(methylsulfinyl)propyl isothiocyanate derivative in the protic solvent increases, compound 2 gradually degrades. At the last sampling, the compound is degraded to less than 60%. Under the same experimental conditions, compounds 2-1, 2-4, and 2-6 are almost not degraded or degraded very little, indicating that their stability is greatly increased compared to compound 2. The structure of compound 2-4 is an aromatic substitution of the thiourea type, and its stability is comparable to that of the aromatic substituted aminodithiocarboxylate compound 2-6. The stability of compounds 2-4 and 2-6 is slightly higher than that of the chain substituted aminodithiocarboxylate compound 2-1. Figure 1 b) It can be found that the above four compounds are relatively stable in aprotic solvents, with little degradation at the final sampling time, and degradation is less than 2%, and the difference is not statistically significant compared with the parent compound 2. In summary, the structural modification and optimization of 3-(methylsulfinyl)propyl isothiocyanate derivatives significantly improves the stability of 3-(methylsulfinyl)propyl isothiocyanate in protic solvents.

[0169] from Figure 1 c) It can be seen that compound 3 gradually degrades as the time of the compound in the protic solvent increases. At the last sampling, compound 3 degraded to less than 20%; under the same experimental conditions, compound 3-1, compound 3-2 and compound 3-5 almost did not degrade or degraded very little, indicating that their stability was greatly increased compared to compound 3. The structure of compound 3-5 is an aromatic substitution of thiourea, and its stability is higher than the structure of the aminodithiocarboxylate compound 3-2 substituted with an aromatic group, and the structural stability of compound 3-2 is higher than the structure of the aminodithiocarboxylate compound 3-1 substituted with a chain hydrocarbon group; Figure 1 d) It can be found that the above four compounds are relatively stable in aprotic solvents, among which the stability of compound 3-2 is reduced compared with that of compound 3, compound 3-1, and compound 3-5, but the overall degradation rate is extremely low, and the degradation rate of the last sampling is less than 5%. The degradation rate of the other three compounds is still that the thiourea aromatic substituted compound 3-5 is better than the chain alkyl substituted aminodithiocarboxylate 3-1, and compound 3-1 is better than the parent compound 3; In summary, the derivatives of 3-(methylsulfonyl)propyl isothiocyanate of the present invention significantly improve the stability of the 3-(methylsulfonyl)propyl isothiocyanate structure in protic solvents through structural modification and optimization.

[0170] 2. Cell Viability Test

[0171] Rat myocardial microvascular endothelial cells (CMECs) were subjected to hypoxia / reoxygenation treatment using a hypoxia / anaerobic workstation Invivo2 400 (Ruskinn, UK), and the effects of the test compounds on their protection against hypoxia / reoxygenation injury were observed.

[0172] Specific operation method:

[0173] (1) Cell culture: The experimental cell line, rat myocardial microvascular endothelial cells (CMECs), was cultured in low-glucose DMEM containing endothelial cell growth supplement (ECGS) (7.5 mg / 500 ml) and 10% fetal bovine serum, and passaged after routine digestion with 0.25% trypsin.

[0174] (2) Establishment and grouping of H / R model: Endothelial cells that had grown and fused into a monolayer at passages 3-5 were randomly divided into four groups: normal control group (Control), hypoxia / reoxygenation group (H / R), experimental group, and positive control drug (Nifedipine, NI) group. Before the experiment, the control group was replaced with fresh 10% FBS low-glucose DMEM complete medium and cultured under conventional culture conditions for 6 hours; the other groups were all subjected to H / R model: after washing the cells twice with PBS, hypoxic solution that had been pre-saturated with high-purity nitrogen for 30 minutes was added. A certain amount of pure nitrogen was filled according to the size of the culture container to expel air, and the cells were quickly placed in a hypoxic box filled with high-purity nitrogen at 37°C for 3 hours; after hypoxia, the cells were washed twice with PBS, replaced with 10% FBS low-glucose DMEM medium (i.e., reoxygenation solution), and cultured under conventional culture conditions for 1 hour to cause reoxygenation damage.

[0175] (3) Detection of rat myocardial microvascular endothelial cell survival rate using Cell Counting Kit-8: Endothelial cells grown into a monolayer at passages 3-5 were digested into single cells and then incubated at 5×10 4 Cells were seeded into 96-well plates at a density of 100 μL / mL, and 100 μL of the diluted cell suspension was added to each well. The cells were cultured in an incubator containing 5% CO2 for 36 h. The experimental group effect was performed according to the experimental steps for establishing and grouping the H / R model. The concentrations of the experimental groups were set as 0.1 μM, 0.5 μM, 1 μM, and 5 μM. The concentrations of the positive control drug nifedipine were set at 1 μM and 5 μM. Each drug concentration corresponded to 3 replicate wells. After the model was established, 10 μM CCK-8 was added to each well. The cells were protected from light and incubated in a 37°C incubator containing 5% CO2 for 2 h. The absorbance value (OD value) at 450 nm was detected with a microplate reader to calculate the cell survival rate.

[0176] The cell survival rate was calculated as the evaluation index using the following method:

[0177] Cell survival rate (%) = [mean OD of experimental group - mean OD of blank group / (mean OD of control group - mean OD of blank group)] × 100%.

[0178] The cell viability test was performed on the 3-(methylsulfinyl)propyl isothiocyanate derivatives and 3-(methylsulfinyl)propyl isothiocyanate in Examples 1 to 7 and the 3-(methylsulfonyl)propyl isothiocyanate derivatives and 3-(methylsulfonyl)propyl isothiocyanate in Examples 8 to 14. The results are as follows: Figure 2 As shown, a) 3-(methylsulfinyl)propyl isothiocyanate derivatives affect the proliferation rate of CMECs cells under H / R; b) 3-(methylsulfonyl)propyl isothiocyanate derivatives affect the proliferation rate of CMECs cells under H / R; #p<0.05versus H / R, ##p<0.01versus H / R*p<0.05versuscontrol, **p<0.01versuscontrol, values ​​are described as mean ± SEM (standard error of the mean, n=3).

[0179] Depend on Figure 2 a) As shown, the survival rate of CMECs in the control group was 100.00%, and the survival rate of CMECs in the H / R group was 50.36%±2.26%. When the concentrations of compound 2-1 were 0.1μM, 0.5μM, 1μM, and 5μM, the survival rates of CMECs were 52.95%±1.83%, 49.62%±5.94%, 53.44%±2.72%, and 44.36%±3.19%, respectively. When the concentrations of compound 2-2 were 0.1μM, 0.5μM, 1μM, and 5μM, the survival rates of CMECs were 51.25%±2.10%, 52.42%±4.59%, 51.07%±1.31%, and 52.84%±2.53%, respectively.

[0180] Depend on Figure 2 a) It can be seen that the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 50.36%±2.26%, and when the concentrations of compounds 2-3 were 0.1μM, 0.5μM, 1μM and 5μM, the survival rates of CMECs were 55.58%±1.86%, 56.49%±0.96%, 70.02%±4.52% and 49.35%±4.20%, respectively; when the concentrations of compounds 2-4 were 0.1μM, 0.5μM, 1μM and 5μM, the survival rates of CMECs were 49.87%±1.58%, 53.41%±0.84%, 48.55%±3.20% and 50.58%±0.84%, respectively.

[0181] Depend on Figure 2a) It can be seen that the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 48.64%±1.64%, and when the concentrations of compounds 2-5 were 0.1μM, 0.5μM, 1μM and 5μM, the survival rates of CMECs were 47.80%±1.67%, 49.76%±4.34%, 46.01%±0.05% and 46.34%±0.61%, respectively; when the concentrations of compounds 2-6 were 0.1μM, 0.5μM, 1μM and 5μM, the survival rates of CMECs were 50.76%±1.22%, 50.00%±0.74%, 48.43%±1.02% and 41.28%±3.19%, respectively.

[0182] Depend on Figure 2 a) As shown, the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 48.64% ± 1.64%, and when the concentrations of compound 2-9 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the survival rates of CMECs were 52.25% ± 1.45%, 53.77% ± 3.93%, 54.15% ± 3.78%, and 58.73% ± 1.56%, respectively. When the concentrations of compound 2 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the survival rates of CMECs were 50.42% ± 2.42%, 50.04% ± 3.25%, 50.20% ± 2.87%, and 48.44% ± 3.64%, respectively.

[0183] Depend on Figure 2 b) As shown, the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 59.97% ± 5.80%, and when the concentrations of compound 3-1 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the survival rates of CMECs were 55.19% ± 5.55%, 60.51% ± 8.84%, 59.87% ± 5.16%, and 58.76% ± 5.80%, respectively. When the concentrations of compound 3-2 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the survival rates of CMECs were 62.31% ± 8.99%, 63.90% ± 7.52%, 67.47% ± 6.01%, and 56.42% ± 6.85%, respectively.

[0184] Depend on Figure 2b) As shown, the CMECs survival rate in the control group was 100.00%, and that in the H / R group was 59.97% ± 5.80%. When the concentrations of compound 3-3 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the CMECs survival rates were 63.31% ± 8.50%, 68.75% ± 6.82%, 73.89% ± 9.30%, and 59.12% ± 11.99%, respectively. When the concentration of compound 3-4 was 0.1 μM, the CMECs survival rate was 59.56% ± 4.63%. When the concentrations of compound 3-4 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, the CMECs survival rates were 59.28% ± 5.24%, 63.75% ± 5.05%, and 56.90% ± 3.73%.

[0185] Depend on Figure 2 b) As shown, the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 53.85% ± 2.14%, and the survival rates of CMECs were 55.10% ± 2.97%, 40.28% ± 3.83%, 47.66% ± 2.86%, and 38.70% ± 2.83% when the concentrations of compound 3-5 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, respectively. The survival rates of CMECs were 54.42% ± 3.52%, 55.28% ± 1.89%, 61.26% ± 3.15%, and 56.60% ± 0.70% when the concentrations of compound 3-6 were 0.1 μM, 0.5 μM, 1 μM, and 5 μM, respectively.

[0186] Depend on Figure 2b) It can be seen that the survival rate of CMECs in the control group was 100.00%, the survival rate of CMECs in the H / R group was 55.32%±1.30%, and when the concentrations of compound 3-9 were 0.1μM, 0.5μM, 1μM and 5μM, the survival rates of CMECs were 55.14%±1.92%, 56.06%±1.18%, 59.18%±1.45%, and 64.50%±1.06%; after modeling, compound 3-9 at 5μM had significant differences in protecting microvascular endothelial cells from hypoxia-reoxygenation injury compared with the modeling group, and the values ​​were statistically significant. When the concentrations of compound 3 were 0.1μM, 0.5μM, 1μM, and 5μM, the survival rates of CMECs were 60.18%±7.82%, 61.41%±6.68%, 61.78%±4.98%, and 60.24%±3.84%. In summary, after modeling, compound 3-3 showed significant differences in protecting against hypoxia-reoxygenation injury in microvascular endothelial cells at a concentration of 1μM, and 0.5μM showed statistically significant protection against hypoxia-reoxygenation injury in microvascular endothelial cells. The survival rate of compound 3-9 at a concentration of 5μM was also significantly different from that of the H / R group, and the comparison was statistically significant at 1μM. Both compounds were significantly superior to the parent compound 3.

[0187] 3. Detection of intracellular ROS levels

[0188] Specific test method: After the endothelial cells that have grown and fused into a monolayer for 3-5 generations are digested into single cells, they are seeded into 6-well plates at a density of 20×104 / mL, 1 mL of diluted cell suspension is added to each well, and cultured in a 5% CO2 incubator for 36 hours; according to the experimental steps for establishing and grouping the H / R model, the action concentration of compound 2-3 is 1 μM, the action concentration of 2-9 is 5 μM; the action concentration of compound 3-3 is 1 μM, the action concentration of 3-9 is 5 μM; the action concentration of NI is 5 μM. After the model is established, DCFH-DA is prepared with serum-free low-glucose DMEM culture medium to a concentration of 5 μM, 1 mL is added to each well for staining, and the cells are incubated in a CO2 incubator for 30 minutes. Fluorescence photography is performed under a phase contrast inverted fluorescence microscope (ZEISS Observer A1) to detect the level of ROS in the cells. The experimental results are as follows. Figure 3 As shown, a) Effects of 3-(methylsulfinyl)propyl isothiocyanate derivatives on reactive oxygen species in CMECs cells; b) Effects of 3-(methylsulfonyl)propyl isothiocyanate derivatives on reactive oxygen species in CMECs cells; **p<0.01versus Control, #p<0.05versus H / R, ##p<0.01versus H / R, values ​​are described as mean ± SEM (standard error of the mean, n=3).

[0189] Depend on Figure 3 a) It can be seen that in CMECs cells damaged by hypoxia and reoxygenation (H / R), the ROS level was significantly increased compared with the control group. When the cells were treated with compounds 2-3, compound 2-9 and the positive control NI group, the intracellular ROS level caused by damage was significantly reduced compared with the H / R group.

[0190] Depend on Figure 3 b) It can be seen that in CMECs cells damaged by hypoxia and reoxygenation (H / R), the ROS level was significantly increased compared with the control group (control). When the cells were treated with compounds 3-3, compound 3-9 and the positive control NI group, the intracellular ROS level caused by damage was significantly reduced compared with the H / R group.

[0191] Obviously, the above embodiments of the present invention are merely examples for the purpose of clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Those skilled in the art will appreciate that other variations or modifications can be made based on the above description. It is not necessary and impossible to enumerate all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the claims of the present invention.

Claims

1. Use of an isothiocyanate derivative in the preparation of a drug for preventing myocardial hypoxia and reoxygenation injury, characterized in that: The structural formula of the isothiocyanate derivative is shown in formula (I): Formula (I) Wherein, X is a sulfoxide group or a sulfone group, R is -S-R1, and R1 is p-tolyl or p-bromobenzyl.

2. The use according to claim 1, characterized in that The isothiocyanate derivative is prepared by the following steps: After fully reacting the reaction monomer I with the reaction monomer II at room temperature, the isothiocyanate derivative can be obtained through extraction, separation and purification; Wherein, the monomer I is or ; The monomer II is HS-R1, and R1 is p-tolyl or p-bromobenzyl.

3. The use according to claim 2, characterized in that The molar ratio of the monomer I to the monomer II is 1:(1-1.5).

4. The use according to claim 2, characterized in that When the monomer II is HS-R1, the monomer I and the monomer II are reacted in an alkaline solution at room temperature for 0.5 to 10 hours to obtain an isothiocyanate derivative.

Images