Hydrogen sulfide prodrug of coumarin compound and application of hydrogen sulfide prodrug

Abstract

The invention discloses a hydrogen sulfide prodrug of a coumarin compound and application of the hydrogen sulfide prodrug, the hydrogen sulfide prodrug of the coumarin compound is shown as a formula (I), the coumarin compound is used as a lead compound, the structure of the coumarin compound is modified and transformed, and release of hydrogen sulfide can be monitored through the fluorescence property of the coumarin compound; according to the invention, release-monitoring is integrated, hydrogen sulfide can be released while the hydrogen sulfide is converted into an active compound in vivo, and the physiological function of the hydrogen sulfide is exerted to regulate oxidative stress of a body and reduce toxic and side effects of the drug while the drug effect of the active compound is exerted, so that the effects of reducing toxicity and enhancing efficacy are achieved. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and particularly relates to a hydrogen sulfide prodrug of a coumarin compound and its application. Background Art

[0002] Coumarin compounds have unique properties and reactivity, and have been utilized in numerous fields. The properties and applications of related compounds have been widely explored in the academic community. This class of compounds is a type of substance with significant biological activity, characterized by a small molecular weight, simple synthesis, high bioavailability, wide pharmacological effects, and low toxicity. They have been widely used in research on anti-tumor, antioxidant, anti-inflammatory, etc. Currently, many coumarin compounds in clinical practice have certain toxic and side effects. For example, the coumarin anticoagulant warfarin is prone to cause serious bleeding tendencies when taken in excess, and thrombosis leading to stroke when taken in small amounts; the cholagogue hydroxymethylcoumarin tablets can cause massive bleeding and cholestatic liver injury when taken in excess; long-term use of the Chinese herbal medicine Psoralea corylifolia can damage the liver and kidney functions. Hydrogen sulfide is a colorless, flammable, highly toxic acidic gas under standard conditions, with a rotten egg smell at low concentrations. Research shows that H2S, similar to CO and NO, is an important gas signaling molecule in the body. Based on the physiological functions of hydrogen sulfide in anti-inflammatory, antioxidant, anti-tumor, and regulating ion channels, introducing hydrogen sulfide is an effective method to achieve reducing toxic and side effects while exerting the physiological functions of hydrogen sulfide itself. However, since hydrogen sulfide gas cannot be directly used for in vivo treatment, how to rationally design H2S prodrugs and enable them to release H2S under appropriate conditions is the key to current research.

[0003] Currently, a variety of hydrogen sulfide prodrugs have been reported, such as the hydrolysis-triggered hydrogen sulfide prodrug demethylthiotriacetate; the thiol-activated hydrogen sulfide prodrug tert-butyl persulfide; the photo-activated hydrogen sulfide prodrug SPD-1; the esterase-triggered hydrogen sulfide prodrug NSAIDs, etc. In addition, hydrogen sulfide detection methods mainly include methylene blue (MB) colorimetry, ion electrode or polarographic electrode methods, gas chromatography (GC-FPD), monobromodiphenyl ether detection methods, and various fluorescence probe methods, etc. The above methods are all independent of release and monitoring, and do not have the ability to monitor H2S in real time in the physiological environment, and there are certain limitations. Summary of the Invention

[0004] Aiming at the problems of existing drugs and hydrogen sulfide "release-monitoring", the purpose of the present invention is to use thioesters as carriers of hydrogen sulfide prodrugs to achieve the integration of "release-monitoring", and exert the physiological functions of hydrogen sulfide. The hydrogen sulfide prodrug design strategy of the present invention has great significance.

[0005] The purpose of the present invention can be achieved by the following measures:

[0006] In one aspect, the present invention provides a hydrogen sulfide prodrug of a coumarin compound as shown in formula (I) or a pharmaceutically acceptable salt thereof.

[0007]

[0008] Wherein: R1, R2 are independently selected from H, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, Or R1 and R2 form with their adjacent carbon atoms

[0009] R3, R4, R5, R6 are independently selected from H, hydroxyl, amino, nitro, cyano,

[0010] Or R3 and R4 form an unsubstituted or R-substituted carbon atom with its adjacent carbon atom. A The substituted aromatic ring is either unsubstituted or R B or R4 and R5 form an unsubstituted or substituted heteroaromatic ring with its adjacent carbon atom. A The substituted aryl group may be unsubstituted or R B or R5 and R6 form an unsubstituted or substituted heteroaromatic ring with its adjacent carbon atom. A The substituted aryl group may be unsubstituted or R B Substituted heteroaromatic ring; the substituent R A , R B Each is independently selected from H, hydroxyl, amino, nitro, cyano, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl;

[0011] m is 0, 1, or 2;

[0012] R a , R b Independently selected from hydroxyl, C1-C6 alkyl.

[0013] In some specific embodiments, R1 is H.

[0014] In some specific examples, R2 is selected from H, hydroxyl, amino, nitro, cyano, hydroxylamine, C1-C4 alkyl, C1-C4 haloalkyl; preferably, R2 is selected from H, hydroxyl, amino, nitro, cyano, methyl, ethyl, n-propyl, isopropyl, C1-C3 haloalkyl,

[0015] In some specific examples, R3 is selected from H.

[0016] In some specific examples, R4 is selected from H, hydroxyl, amino, nitro, and cyano.

[0017] In some specific examples, R5 is selected from H, hydroxyl, amino, nitro, cyano,

[0018] In some specific examples, R6 is selected from H.

[0019] In some specific examples, the aromatic ring in the present invention is a benzene ring or a naphthalene ring, preferably a benzene ring; in some examples, R A is preferably H, hydroxyl, amino, nitro, methyl, ethyl, methoxy, ethoxy, trifluoromethyl.

[0020] In some specific examples, the heteroaromatic ring in the present invention is a 5- to 6-membered heteroaromatic ring having 1 to 3 selected from N, O or S, preferably, the heteroaromatic ring is selected from furan ring, thiophene ring, pyrrole ring, pyrazole ring, imidazole ring, oxazole ring, thiazole ring, pyridine ring, pyrimidine ring; in some examples, R B is preferably H, hydroxyl, amino, nitro, methyl, ethyl, methoxy, ethoxy, trifluoromethyl; more preferably, R B is H.

[0021] In some specific examples, m is 0 or 1.

[0022] In some specific examples, R a and R b are independently selected from hydroxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0023] On the one hand, the present invention provides a hydrogen sulfide prodrug represented by formula (II) or a pharmaceutically acceptable salt thereof,

[0024]

[0025] wherein, R is selected from hydroxyl or C1-C6 alkyl; preferably, R is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0026] The present invention also provides hydrogen sulfide prodrugs of some specific coumarin compounds represented by the following structures or pharmaceutically acceptable salts thereof:

[0027]

[0028]

[0029] The hydrogen sulfide prodrug of the coumarin compound described in the present invention can release hydrogen sulfide and the original drug under the action of in vivo enzymes, thereby exerting the physiological function of hydrogen sulfide, increasing the efficacy of the drug and reducing its toxic and side effects.

[0030] The enzyme described in the present invention may include any enzyme that can directly hydrolyze an ester bond or a thioester and directly release H2S or release hydrogen sulfide through chemical allosterism. Through various in vivo and in vitro experiments, the inventors found that a variety of enzymes in the body can achieve this function.

[0031] Mechanism of the hydrogen sulfide prodrug of the coumarin compound shown in formula (I) of the present invention:

[0032]

[0033] Mechanism of the hydrogen sulfide prodrug of the compound shown in formula (II) of the present invention:

[0034]

[0035] The present invention also provides a preparation method of the coumarin compound shown in formula (I):

[0036]

[0037] The present invention also provides a preparation method of the hydrogen sulfide prodrug of the compound shown in formula (II):

[0038]

[0039] The present invention also provides a pharmaceutical composition, comprising the hydrogen sulfide prodrug described in the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0040] The hydrogen sulfide prodrug described in the present invention can be administered to a subject with the compound of the present invention or a pharmaceutical composition containing the composition of the present invention in an environment sufficient to form hydrogen sulfide or persulfide and the parent drug.

[0041] The present invention also provides the use of the hydrogen sulfide prodrug described above in the preparation of a drug for treating stroke; especially ischemic stroke.

[0042] The compound of the present invention or a pharmaceutically acceptable salt thereof can be used alone as a single therapeutic drug or in combination with other active drugs.

[0043] Advantages of the present invention:

[0044] The present invention uses coumarin compounds as lead compounds, modifies and transforms their structures, and can monitor the release of hydrogen sulfide through their own fluorescence properties. It not only integrates "release - monitoring" but also releases hydrogen sulfide while being transformed into the parent drug in vivo. While exerting the efficacy of the parent drug, it exerts the physiological function of hydrogen sulfide to regulate the body's oxidative stress and reduce the drug's toxic and side effects, achieving the effect of reducing toxicity and increasing efficacy. Brief Description of the Drawings

[0045] Figure 1 Pharmacodynamic evaluation of ZYSO02 (A: infarct area; B: analysis of serum LDH level; C: analysis of MDA level in brain tissue);

[0046] Figure 2 Pharmacodynamic evaluation of ZYSO03 (A: infarct area; B: analysis of serum LDH level; C: analysis of MDA level in brain tissue);

[0047] Figure 3 Determination of serum biochemical indexes 24 hours after administration of high-dose Psoralen (800 mg / kg, i.p.), high-dose ZYSO03 (869 mg / kg, i.p.), low-dose Psoralen (400 mg / kg, i.p.), and low-dose ZYSO03 (435 mg / kg, i.p.) (A, B: organ index; C, E: serum ALT level; D, F: serum AST level; G, H: serum ALP level; I, J: serum TP level; K, L: γ-GT level in liver tissue; M, N: GSH level in kidney tissue; O, P: serum ALB level);

[0048] Figure 4 Anticoagulation evaluation of ZYSO02 and ZYSO03 (A: bleeding time; B: prothrombin time; C: activated partial thromboplastin time);

[0049] Figure 5 Animal fluorescence imaging results of ZYSO20. Detailed implementation manners

[0050] The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples are all obtained from regular biochemical reagent stores unless otherwise specified.

[0051] Example 1: Compound ZYSO01

[0052]

[0053] Coumarin (1.46 g, 10 mmol) and Lawesson's reagent (8.09 g, 20 mmol, 2 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After purging with N2, toluene (50 mL) was added by syringe as the solvent, and the mixture was refluxed and stirred at 110 °C for 12 h. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) to give the yellow solid ZYSO01 (1 g, 60%).

[0054] 1 H NMR (300 MHz, CDCl3) δ 7.59 - 7.47 (m, 2H), 7.42 (dd, J = 8.7, 3.1 Hz, 2H), 7.34 - 7.26 (m, 1H), 7.18 (d, J = 9.3 Hz, 1H).

[0055] 13 C NMR (300 MHz, CDCl3) δ 198.00, 156.62, 134.66, 132.28, 129.64, 127.91, 125.57, 120.45, 116.73

[0056] Example 2 Compound ZYSO02

[0057]

[0058] 1-1A (1.76 g, 10 mmol), tert-butyldimethylchlorosilane (2.26 g, 15 mmol, 1.5 eq), and imidazole (2.04 g, 30 mmol, 3 eq) were added to a 250 mL two-necked flask. After purging with N2, anhydrous DMF (50 mL) was added by syringe as the solvent, and the mixture was stirred overnight at room temperature. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with PE (30 mL × 3). The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 8:1) to give the white solid 1-2A (2.67 g, 92%).

[0059] 1-2A (290 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After purging with N2, toluene (5 mL) was added as a solvent using a syringe, and the mixture was refluxed and stirred at 110 °C for 6 h. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) and eluted to obtain yellow solid 1-3A (287 mg, 94%).

[0060] 1-3A (306 mg, 1.0 mmol) and tetrabutylammonium fluoride trihydrate (694 mg, 2.2 mmol, 2.2 eq) were added to an eggplant-shaped flask containing a magnetic stir bar. Under an ice bath, THF (5 mL) was added as a solvent using a syringe, and the mixture was stirred at room temperature for 12 h. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) and eluted to obtain yellow solid ZYSO02 (162 mg, 84%).

[0061] 1 H NMR (300 MHz, DMSO-d6) δ 10.87 (s, 1H), 7.70 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 0.9 Hz, 1H), 6.90 (dt, J = 4.9, 2.2 Hz, 2H), 2.34 (d, J = 0.8 Hz, 3H).

[0062] 13 C NMR (300 MHz, DMSO-d6) δ 196.30, 161.96, 157.52, 147.12, 126.94, 124.90, 114.74, 113.85, 101.85, 17.53.

[0063] ESI-HRMS: calcd. for C 10 H8O2S [M+H] + 193.23, found 193.03279.

[0064] Example 3 Compound ZYSO03

[0065]

[0066] With reference to the synthesis of ZYSO01, psoralen (186 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain a yellow solid ZYSO03 (100 mg, 50%).

[0067] 1 H NMR (300 MHz, CDCl3) δ 7.77 - 7.68 (m, 2H), 7.62 (s, 1H), 7.54 (d, J = 9.3 Hz, 1H), 7.18 (d, J = 9.3 Hz, 1H), 6.84 (dd, J = 2.2, 0.9 Hz, 1H).

[0068] 13 C NMR (300 MHz, CDCl3) δ 198.10, 156.66, 154.57, 147.73, 135.53, 127.77, 126.01, 119.72, 117.06, 106.56, 99.82.

[0069] ESI-HRMS: calcd. for C 11 H6O2S [M+H] + 203.23, found 203.01579.

[0070] Example 4 Compound ZYSO04

[0071]

[0072] With reference to the synthesis of ZYSO02, aesculetin (1.78 g, 10 mmol) was used as the starting material to synthesize a yellow solid ZYSO04 (162 mg, 84%).

[0073] 1 H NMR (300 MHz, DMSO-d6) δ 10.21 (s, 2H), 7.75 (d, J = 9.2 Hz, 1H), 7.05 (d, J = 9.4 Hz, 2H), 6.95 (s, 1H).

[0074] 13 C NMR (300 MHz, DMSO-d6) δ 196.12, 152.26, 151.87, 144.53, 136.96, 125.19, 113.30, 111.65, 102.36

[0075] Example 5 Compound ZYSO05

[0076]

[0077] For the synthesis of ZYSO01, 7-hydroxycoumarin (160 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain yellow solid ZYSO05 (70 mg, 40%).

[0078] 1 H NMR (300 MHz, DMSO-d6) δ 10.93 (s, 1H), 7.80 (d, J = 8.9 Hz, 1H), 7.67 - 7.62 (m, 1H), 7.06 (d, J = 9.2 Hz, 1H), 6.93 - 6.87 (m, 2H).

[0079] 13 C NMR (300 MHz, DMSO-d6) δ 197.19, 162.24, 158.15, 136.99, 130.05, 125.21, 115.03, 113.30, 101.90

[0080] Example 6 Compound ZYSO06

[0081]

[0082] For the synthesis of ZYSO01, 7-amino-4-methylcoumarin (175 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain yellow solid ZYSO06 (90 mg, 46%).

[0083] 1 H NMR (300 MHz, CDCl3) δ 7.36 (d, J = 9.0 Hz, 1H), 6.58 (dd, J = 7.8, 1.8 Hz, 2H), 6.02 (d, J = 1.0 Hz, 1H), 4.16 (s, 2H), 2.35 (d, J = 1.0 Hz, 3H).

[0084] 13 C NMR (300 MHz, CDCl3) δ 195.46, 158.43, 154.02, 147.81, 126.67, 122.39, 113.22, 110.98, 97.61, 17.43

[0085] Example 7 Compound ZYSO07

[0086]

[0087] For the synthesis of ZYSO01, 7-hydroxy-4-trifluoromethylcoumarin (229 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain a yellow solid ZYSO07 (40 mg, 16%).

[0088] 1 H NMR (300 MHz, CH3OD) δ 7.65 - 7.59 (m, 1H), 7.27 (d, J = 0.7 Hz, 1H), 6.92 (q, J = 2.3 Hz, 2H).

[0089] 13 C NMR (300 MHz, CH3OD) δ 197.11, 164.25, 127.41, 124.82, 116.21, 111.64, 108.84, 104.37, 103.92, 101.39

[0090] Example 8 Compound ZYSO08

[0091]

[0092] For the synthesis of ZYSO01, 7-dimethylamino-4-methylcoumarin (203 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain a yellow solid ZYSO08 (40 mg, 20%).

[0093] 1 H NMR (300 MHz, CDCl3) δ 7.46 (d, J = 8.7 Hz, 1H), 6.95 (s, 1H), 6.73 - 6.65 (m, 2H), 3.08 (s, 6H), 2.30 (s, 3H).

[0094] Example 9 Compound ZYSO09

[0095]

[0096] For the synthesis of ZYSO01, 7-dimethylamino-4-methylcoumarin (230 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain a yellow solid ZYSO09 (49 mg, 20%).

[0097] 11H NMR (300 MHz, DMSO-d6) δ 7.48 - 7.41 (m, 1H), 7.08 (s, 1H), 6.86 (s, 2H), 6.77 (dd, J = 8.9, 2.2 Hz, 1H), 6.65 (d, J = 2.1 Hz, 1H).

[0098] 13 13C NMR (300 MHz, DMSO-d6) δ 194.87, 159.58, 154.83, 132.45, 132.03, 125.95, 119.26, 114.16, 104.19, 98.34

[0099] Example 10 Compound ZYSO10

[0100]

[0101] Referring to the synthesis of ZYSO01, isopsoralen (186 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain yellow solid ZYSO10 (100 mg, 50%).

[0102] 1 1H NMR (300 MHz, CDCl3) δ 7.81 (d, J = 9.6 Hz, 1H), 7.69 (d, J = 2.2 Hz, 1H), 7.41 (dt, J = 17.7, 4.7 Hz, 2H), 7.12 (dd, J = 2.2, 0.8 Hz, 1H), 6.39 (d, J = 9.6 Hz, 1H).

[0103] 13 13C NMR (300 MHz, CDCl3) δ 197.40, 157.58, 151.32, 146.33, 136.01, 128.14, 123.80, 116.71, 115.71, 110.23, 104.50

[0104] Example 11 Compound ZYSO11

[0105]

[0106] 7-Methoxy-2-naphthol (200 mg, 1.2 mmol, 1 eq) was suspended in concentrated sulfuric acid (1.5 mL). After adding ethyl 4-chloroacetoacetate (365 μL, 2.5 mmol, 2.5 eq) at 0 °C, the mixture was stirred at room temperature for 24 h. After quenching with ice water, water (10 mL) was added and the mixture was extracted with DCM (30 mL × 3). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 5:1) and eluted to obtain the yellow solid 1-1C (100 mg, 30%).

[0107] 1-1C (274 mg, 1.0 mmol, 1 eq) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After displacing N2, toluene (5 mL) was added as a solvent using a syringe, and the mixture was refluxed and stirred at 110 °C for 1 h. The system changed from turbid to clear, and TLC monitoring showed that the reaction was basically complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added and the mixture was extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) and eluted to obtain the yellow solid ZYS011 (174 mg, 60%).

[0108] 1 H NMR (300 MHz, DMSO-d6) δ 8.25 (d, J = 8.9 Hz, 1H), 8.04 (s, 1H), 7.92 (d, J = 1.8 Hz, 1H), 7.65 - 7.55 (m, 2H), 7.36 (dd, J = 8.9, 2.2 Hz, 1H), 5.40 (s, 2H), 3.99 (s, 3H).

[0109] Example 12 Compound ZYSO12

[0110]

[0111] In a 250 mL two-necked flask, 2,7-dihydroxynaphthalene (1.6 g, 10 mmol, 1 eq), tert-butyldimethylchlorosilane (2.26 g, 15 mmol, 1.5 eq), and imidazole (2.04 g, 30 mmol, 3 eq) were added. After replacing N₂, anhydrous DMF (50 mL) was added by syringe as the solvent, and the mixture was stirred overnight at room temperature. The system changed from turbid to clear, and TLC monitoring showed that the reaction was basically complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with PE (30 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 8:1) and eluted to obtain white solid 1-2D (1.1 g, 40%).

[0112] 1-2D (274 mg, 1.0 mmol, 1 eq) was added to ethyl 4-chloroacetoacetate (380 μL, 2.5 mmol, 2.5 eq) at 0 °C. After 30 min, concentrated sulfuric acid (1 mL) was added, and the mixture was stirred at room temperature for 24 h. After quenching with ice water, water (10 mL) was added, and the mixture was extracted with DCM (30 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 5:1) and eluted to obtain yellow solid 1-3D (68 mg, 26%).

[0113] 1-3D (260 mg, 1.0 mmol, 1 eq) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After replacing N₂, toluene (5 mL) was added by syringe as the solvent, and the mixture was refluxed and stirred at 110 °C for 1 h. The system changed from turbid to clear, and TLC monitoring showed that the reaction was basically complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) and eluted to obtain yellow solid ZYSO12 (193 mg, 70%).

[0114] 1 H NMR (300 MHz, DMSO-d6) δ 10.24 (s, 1H), 8.09 (d, J = 8.9 Hz, 1H), 7.92 (d, J = 8.8 Hz, 1H), 7.82 (d, J = 2.0 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.17 (dd, J = 8.8, 2.1 Hz, 1H), 6.77 (s, 1H), 5.30 (s, 2H).

[0115] Example 13 Compound ZYSO13

[0116]

[0117] With reference to the synthesis of ZYSO11, yellow solid ZYSO13 (25 mg, 10%) was synthesized using 7-methoxy-2-naphthol (200 mg, 1.2 mmol, 1 eq) and ethyl acetoacetate (365 μL, 2.5 mmol, 2.5 eq) as starting materials.

[0118] 1 H NMR (300 MHz, CDCl3) δ 8.04 (s, 1H), 7.82 (d, J = 9.1 Hz, 1H), 7.73 (s, 1H), 7.17 (dd, J = 9.1, 2.4 Hz, 1H), 7.13 (s, 2H), 3.97 (s, 3H), 2.46 (d, J = 0.9 Hz, 3H).

[0119] Example 14 Compound ZYSO14

[0120]

[0121] With reference to the synthesis of ZYSO11, yellow solid ZYSO14 (25 mg, 10%) was synthesized using 7-methoxy-2-naphthol (200 mg, 1.2 mmol, 1 eq) and ethyl acetoacetate (365 μL, 2.5 mmol, 2.5 eq) as starting materials.

[0122] 1 H NMR (300 MHz, CDCl3) δ 7.93 (d, J = 9.1 Hz, 2H), 7.83 (d, J = 9.0 Hz, 1H), 7.45 (d, J = 8.8 Hz, 1H), 7.24 (s, 2H), 3.95 (s, 3H), 2.86 (s, 3H).

[0123] Example 15 Compound ZYSO15

[0124]

[0125] With reference to the synthesis of ZYSO12, yellow solid ZYSO15 (25 mg, 10%) was synthesized using 2,7-dihydroxynaphthalene (1.6 g, 10 mmol, 1 eq) as the starting material.

[0126] 1 H NMR (300 MHz, DMSO-d6) δ 10.35 (s, 1H), 8.38 (s, 1H), 8.01 (s, 1H), 7.83 (s, 1H), 7.21 (s, 1H), 7.17 (s, 2H), methyl overlapping DMSO ∼ 2.5 (3H).

[0127] Example 16 Compound ZYSO16

[0128]

[0129] Referring to the synthesis of ZYSO12, using 2,7-dihydroxynaphthalene (1.6 g, 10 mmol, 1 eq) as the raw material, yellow solid ZYSO16 (25 mg, 10%) was synthesized.

[0130] 1 H NMR (300 MHz, DMSO-d6) δ 10.31 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 8.06 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.47 (d, J = 8.9 Hz, 1H), 7.34 (s, 1H), 7.20 (dd, J = 8.8, 2.1 Hz, 1H), 2.84 (s, 3H).

[0131] Example 17 Compound ZYSO17

[0132]

[0133] 4-Hydroxycoumarin (2 g, 12 mmol), benzylideneacetone (3.6 g, 24 mmol, 2 eq), and triethylbenzylammonium chloride (420 mg, 1.8 mmol, 0.15 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After replacing N2, water (60 mL) was added by syringe as the solvent, and the mixture was refluxed and stirred at 100 °C for 7 h. The system changed from turbid to clear, and TLC monitoring showed that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1) and eluted to obtain white solid warfarin (2.4 g, 65%).

[0134] Warfarin (3.08 g, 10 mmol), tert-butyldimethylchlorosilane (2.26 g, 15 mmol, 1.5 eq), and imidazole (2.04 g, 30 mmol, 3 eq) were added to a 250 mL two-necked flask. After replacing N2, anhydrous DMF (3 mL) was added by syringe as the solvent, and the mixture was stirred at 80 °C for 24 h. The system changed from turbid to clear, and TLC monitoring showed that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with PE (30 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 8:1) and eluted to obtain white solid 1-1g-TBS (1.8 g, 43%).

[0135] 1-1g-TBS (422 mg, 1 mmol) and Lawesson's reagent (809 mg, 2 mmol, 2 eq) were added to a two-necked round-bottom flask containing a magnetic stir bar. After purging with N2, toluene (5 mL) was added as a solvent using a syringe, and the mixture was refluxed and stirred at 110 °C for 12 h. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure. The crude product obtained after concentration was directly used in the next step. Tetrabutylammonium fluoride trihydrate (694 mg, 2.2 mmol, 2.2 eq) was added to an eggplant-shaped flask containing a magnetic stir bar. THF (5 mL) was added as a solvent using a syringe under an ice bath, and the mixture was stirred at room temperature for 12 h. The system changed from turbid to clear, and TLC monitoring indicated that the reaction was almost complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with EA (30 mL × 3). The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1), and elution gave a yellow solid ZYSO17 (40 mg, 10%).

[0136] 1 H NMR (300 MHz, DMSO-d6) δ 7.86 (t, J = 6.9 Hz, 1H), 7.73 (t, J = 7.8 Hz, 1H), 7.58 (d, J = 8.2 Hz, 1H), 7.48 (t, J = 7.6 Hz, 1H), 7.16 (ddd, J = 16.5, 14.4, 7.0 Hz, 5H), 4.91 (dd, J = 13.6, 5.4 Hz, 1H), 2.56 (d, J = 6.5 Hz, 1H), 2.37 (dd, J = 14.1, 8.7 Hz, 1H), 1.68 (s, 1H), 1.47 (s, 2H).

[0137] 13 C NMR (300 MHz, DMSO-d6) δ 192.62, 153.66, 147.43, 143.81, 132.99, 130.15, 128.79, 128.36, 128.16, 128.06, 126.34, 126.09, 124.44, 119.88, 116.86, 82.72, 48.42, 43.45, 30.84.

[0138] Example 18 Compound ZYSO18

[0139]

[0140] For the synthesis of ZYSO01, methyl benzoate (136 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain the yellow solid ZYSO18 (75 mg, 50%).

[0141] 1 H NMR (300 MHz, CDCl3) δ 8.23 - 8.18 (m, 2H), 7.58 - 7.51 (m, 1H), 7.40 (dd, J = 10.9, 4.5 Hz, 2H), 4.31 (s, 3H).

[0142] 13 C NMR (75 MHz, CDCl3) δ 212.40, 138.31, 132.84, 128.87, 128.20, 59.43, 29.83.

[0143] Example 19 Compound ZYSO20

[0144]

[0145] For the synthesis of ZYSO01, 7 - diethylamino - 4 - methylcoumarin (231 mg, 1.0 mmol) and Lawesson's reagent (809 mg, 2.0 mmol, 2 eq) were refluxed in toluene at 110 °C to obtain the yellow solid ZYSO20 (120 mg, 50%).

[0146] 1 H NMR (400 MHz, DMSO - d6) δ 7.58 (d, J = 9.1 Hz, 1H), 6.89 - 6.78 (m, 2H), 6.67 (d, J = 2.4 Hz, 1H), 3.45 (q, J = 7.0 Hz, 4H), 2.30 (s, 3H), 1.12 (t, J = 7.0 Hz, 6H).

[0147] 13 C NMR (400 MHz, DMSO - d6) δ 196.01, 158.94, 151.41, 147.89, 126.97, 122.96, 111.02, 110.86, 96.46, 44.64, 44.64, 17.79, 12.77, 12.77.

[0148] Example 20 Compound ZYSO21

[0149]

[0150] 1-1R (406 mg, 1.0 mmol) was added to a 100 mL eggplant-shaped flask, and then CF3COOH and DCM were added to the flask in a ratio of 1:3 as solvents. The reaction was carried out at room temperature for 20 h, and the reaction was monitored by TLC and was basically complete. The solvent was removed by distillation under reduced pressure, water (10 mL) was added, and the mixture was extracted with PE (30 mL * 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The crude product obtained after concentration was separated by silica gel column chromatography (mobile phase: PE:EA = 4:1), and elution gave a dark brown solid ZYSO21 (120 mg, 50%).

[0151] 1 H NMR (400 MHz, DMSO-d6) δ 8.16 (s, 1H), 7.71 (d, J = 9.1 Hz, 1H), 7.01 (s, 1H), 6.82 (dd, J = 9.2, 2.5 Hz, 1H), 6.70 (d, J = 2.5 Hz, 1H), 3.81 (s, 2H), 3.47 (d, J = 7.0 Hz, 4H), 2.64 (s, 3H), 1.13 (t, J = 7.0 Hz, 6H).

[0152] Activity experiment

[0153] Determination of the enzymatic hydrolysis half-life of the compound in Example 21

[0154] 1. Detection of PLE hydrolysis of ZYSO series thioesters and formation of corresponding esters by high performance liquid chromatography

[0155] The ZYSO series solution (100 μL, 500 μM), PBS buffer (4900 μL, pH = 7.4) and PLE (1 mg) were added to a 5 mL sample vial. After sealing with a sealing film, the vial was incubated in a 37 °C water bath. At regular intervals, 200 μL of the solution was taken and transferred to a 2 mL centrifuge tube containing 200 μL of chromatographic methanol. After centrifugation at 13000 rpm for 5 min, the solution was filtered through a 0.22 μm filter membrane and analyzed by HPLC mobile phase elution (chromatographic column: Agilent, 4.6 mm × 150 mm; mobile phase: ACN acetonitrile (A) and 0.01% HCOOH formic acid / water H2O (B); flow rate: 1.0 mL / min; injection volume: 5 μL. Linear gradient elution: 0 - 15 min, 10% - 90% A.) to analyze the components in the above solution. The original drug released at each time point was calculated according to the standard curve, and the release half-life was calculated therefrom.

[0156] 2. Detection of plasma hydrolysis of ZYSO series thioesters and formation of corresponding esters by enzyme-linked immunosorbent assay

[0157] Add blank rat plasma (100 μL), Tris-HCl buffer solution (350 μL, 50 mmol / L, pH = 7.4), and 20 μL of the ZYSO series stock solution (300 μg / mL, methanol) to a 5-mL sample vial to start the reaction. Incubate the mixture at 37 °C, terminate the reaction using saturated EDTA-2Na solution (80 μL), and pipette 150 μL of the solution into a 96-well plate. Measure the fluorescence intensity at a specific wavelength, calculate the amount of the original drug released at each time point according to the standard curve, and thereby calculate its release half-life.

[0158] The results are shown in Table 1, indicating that the formation of the relevant original drugs can be detected for all the above-mentioned compounds, and fluorescence can be basically released at a certain rate to generate hydrogen sulfide under the conditions of esterase and plasma.

[0159] Table 1 Thioester structures, names, and half-lives of enzymatic hydrolysis in the examples of this patent

[0160]

[0161]

[0162] Among them, for the compounds with a half-life greater than 120 h, it does not mean no release. It was found during the detection process that all of them had the behavior of releasing H2S, but the release rate was extremely slow (only 120 h was detected in this experiment), and there is a certain development prospect for slow monitoring.

[0163] Example 22 Pharmacodynamic study in an ischemia-reperfusion injury model

[0164] 1. Establishment of an animal ischemia-reperfusion injury model:

[0165] Male C57BL / 6JNifdc mice weighing 25 - 30 g were randomly divided into 5 groups: sham operation group, model group, and different drug administration groups, with 8 mice in each group. The sham operation group was not inserted with a suture, and the middle cerebral artery occlusion model was applied to the remaining groups. Anesthesia was induced with 3% isoflurane and maintained with 0.6% isoflurane after anesthesia. After disinfection, a median incision was made in the neck. Blunt dissection was performed to expose the left common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). The proximal end of the CCA was separated from the distal end of the ECA and ligated. The CCA was ligated distally, and a suture (Beijing Xinnong Biotechnology Co., Ltd.) was inserted and passed through the ICA until the middle cerebral artery (MCA) was blocked. We first made the brains of C57BL / 6JNifdc male mice ischemic for 1 h 20 min, and injected the corresponding drugs at a dose of 100 μmol / kg via the tail vein during ischemia, and then reperfused for 24 h. Other treatments were the same as above. Blood was collected after 24 h, and after blood collection, the mice were sacrificed by spinal cord transection, and the brain tissues were taken for further evaluation. First, the infarct area of the brain tissue was evaluated by TTC staining; subsequently, we further measured the serum LDH level and the brain tissue MDA level to evaluate the drug efficacy.

[0166] 2. Activity data of compound ZYSO02

[0167] (1) Determination of infarct area

[0168] The mouse brains were frozen at -20 °C for 20 min and cut into 5 pieces, each 1 mm thick. Each slice was immediately stained with 1% 2,3,5-triphenyltetrazolium chloride (TTC) solution at 37 °C for 30 min and fixed in 4% paraformaldehyde (PFA) solution. The infarcted area was not stained, and the normal area was stained red. Then, each pair of slices was photographed, and the cerebral infarct area was calculated by ImageJ. The formula is as follows: infarct volume = sum of infarct areas / total area ( Figure 1 A). The research results showed that after administration of 100 μmol / kg ZYSO02, the drug efficacy was significantly better than that of hymecromone after equimolar administration.

[0169] (2) Determination of serum LDH level and brain tissue MDA level

[0170] After blood collection, serum was collected by centrifugation at 3000 g for 10 min at 4 °C. The method for measuring LDH is described in the kit (solarbio, Beijing) instruction manual. Brain tissues were taken to measure the MDA level in the tissues, and the measurement method is described in the MDA kit (solarbio, Beijing) instruction manual. The LDH level reflects the damage of cell membranes caused by ischemia-reperfusion injury, and the MDA level reflects the peroxidation of membrane lipids caused by ischemia-reperfusion injury. The study found that ZYSO02 showed greater potential in the MCAO model after administration of 100 μmol / kg, such as Figure 1B, Figure 1 C.

[0171] (3) Test of PLE enzymatic hydrolysis of ZYSO series compounds

[0172] Taking the release of the original drug hymecromone by ZYSO02 as an example. Add ZYSO02 solution (100 μL, 500 μM), PBS buffer (4900 μL, pH = 7.4) and PLE (1 mg) into a 5 mL sample bottle. After sealing with a sealing film, incubate it in a 37 °C water bath. At regular intervals, take 200 μL of the solution each time, filter it through a 0.22 μm filter membrane, and then analyze the components in the above solution using a suitable HPLC mobile phase and elution (chromatographic column: Agilent, 4.6 mm × 150 mm; mobile phase: ACN acetonitrile (A) and 0.01% HCOOH formic acid / water H2O (B); flow rate: 1.0 mL / min; injection volume: 5 μL. Linear gradient elution: 0 - 15 min, 10% - 90% A.). Calculate the hymecromone released at each time point according to the standard curve of hymecromone, and calculate its release t 1 / 2 It was 57.59 ± 0.15 h.

[0173] (4) Test of plasma enzymatic hydrolysis of ZYSO series compounds

[0174] Taking the release of the original drug hymecromone by ZYSO02 as an example. Add blank rat plasma (100 μL), Tris - HCl buffer (350 μL, 50 mM, pH = 7.4) and 20 μL of the drug mother liquor (300 μg / mL, methanol) into a 5 mL sample bottle to start the reaction. Incubate the mixture at 37 °C, terminate the reaction using saturated EDTA - 2Na solution (80 μL), and pipette 150 μL of the solution into a 96 - well plate. Measure the fluorescence intensity at an excitation wavelength of 360 nm and an emission wavelength range of 400 - 600 nm. Calculate the hymecromone released at each time point according to the standard curve of hymecromone, and calculate its release t 1 / 2 It was 2.52 ± 0.13 h.

[0175] (5) Determination of coagulation function - related indicators

[0176] Select C57BL / 6JNifdc male mice, randomly divide them into 3 groups: Normal group, hymecromone group (40 mg / kg), and ZYSO02 group (44 mg / kg). Administer the drugs again 6 h after intraperitoneal administration. Then, make an incision with a blade 2.5 mm away from the tail, adsorb the blood with filter paper every 30 s until the blood stops oozing, and record the bleeding time ( Figure 4 A); In addition, after anesthesia with isoflurane, collect blood from the eyeballs, and measure the prothrombin time and activated partial thromboplastin time ( Figure 4B, C), and the measurement method is described in the instruction manual of the kit (Regen Biotech).

[0177] 3. Activity data of compound ZYSO03

[0178] (1) Measurement of infarct area

[0179] The mouse brain was frozen at -20 °C for 20 min and cut into 5 pieces, each 1 mm thick. Each slice was immediately stained with 1% 2,3,5-triphenyltetrazolium chloride (TTC) solution at 37 °C for 30 min and fixed in 4% paraformaldehyde (PFA) solution. The infarcted area was unstained, and the normal area was stained red. Then, each pair of slices was photographed, and the cerebral infarct area was calculated using ImageJ. The formula is as follows: infarct volume = sum of infarct areas / total area ( Figure 2 A). The research results showed that after administration of 100 μmol / kg ZYSO03, the drug efficacy was significantly better than that of psoralen after equimolar administration.

[0180] (2) Measurement of serum LDH level and brain tissue LDA level

[0181] After blood collection, the serum was collected by centrifugation at 3000 g for 10 min at 4 °C. The measurement method of LDH is described in the instruction manual of the kit (Solarbio, Beijing). The brain tissue was taken to measure the MDA level, and the measurement method is described in the instruction manual of the MDA kit (Solarbio, Beijing). The LDH level reflects the damage of cell membranes caused by ischemia-reperfusion injury, and the MDA level reflects the peroxidation of membrane lipids caused by ischemia-reperfusion injury. It was found that ZYSO03 showed greater potential in the MCAO model after administration of 100 μmol / kg, such as Figure 2 B, Figure 2 C.

[0182] (3) Acute liver and kidney injury of ZYSO03

[0183] Male C57BL / 6JNifdc mice were selected and fasted for 12 h before drug administration while having normal access to water. The mice were randomly divided into 5 groups, namely the normal saline control group, the high-dose Psoralen group, the high-dose ZYSO03 group, the low-dose Psoralen group, and the low-dose ZYSO03 group. The administration dose of the high-dose Psoralen group was 800 mg / kg, and the high-dose ZYSO03 group was administered an equimolar dose. The administration dose of the low-dose Psoralen group was 400 mg / kg, and the low-dose ZYSO03 group was administered an equimolar dose. There were 6 mice in each group. The administration method was intraperitoneal injection, and the administration cycle was 24 h. After 24 h, blood was collected from the eye socket and the mice were sacrificed by cervical dislocation. The organ indices and liver toxicity such as serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP), and gamma-glutamyl transpeptidase (γ-GT) were measured; the kidney toxicity such as albumin (ALB) and reduced glutathione (GSH) levels ( Figure 3 A-P).

[0184] (4) Testing of the enzymatic hydrolysis of ZYSO series compounds by PLE

[0185] Taking the release of the original drug Psoralen from ZYSO03 as an example. Add ZYSO03 solution (100 μL, 500 μM), PBS buffer (4900 μL, pH = 7.4), and PLE (1 mg) into a 5 mL sample vial. Seal it with a sealing film and incubate it in a 37 °C water bath. At regular intervals, take 200 μL of the solution each time, filter it through a 0.22 μm filter membrane, and then analyze the components in the above solution using a suitable HPLC mobile phase and elution (Chromatographic column: Agilent, 4.6 mm × 150 mm; Mobile phase: ACN acetonitrile (A) and 0.01% HCOOH formic acid / water H2O (B); Flow rate: 1.0 mL / min; Injection volume: 5 μL. Linear gradient elution: 0 - 15 min, 10% - 90% A.). Calculate the amount of Psoralen released at each time point according to the standard curve of Psoralen, and then calculate its release t 1 / 2 was 62.03 h.

[0186] (5) Testing of the enzymatic hydrolysis of ZYSO series compounds by plasma

[0187] Taking the release of the original drug Psoralen by ZYSO03 as an example. Add blank rat plasma (100 μL), Tris-HCl buffer solution (350 μL, 50 mmol / L, pH = 7.4), and 20 μL of the drug mother liquor (300 μg / mL, methanol) to a 5 mL sample bottle to start the reaction. Incubate the mixture at 37 °C, terminate the reaction with saturated EDTA-2Na solution (80 μL), and pipette 150 μL of the solution into a 96-well plate. Measure the fluorescence intensity at an excitation wavelength of 360 nm and an emission wavelength of 400 - 600 nm. Calculate the Psoralen released at each time point according to the standard curve of Psoralen, and calculate its release t 1 / 2 was 3.95 ± 0.45 h.

[0188] (6) Determination of coagulation function-related indicators

[0189] Select C57BL / 6JNifdc male mice, randomly divide them into 3 groups: Normal group, Psoralen group (40 mg / kg), and ZYSO03 group (44 mg / kg). Administer the drug intraperitoneally again 6 h after the first administration. Then, make an incision with a blade 2.5 mm away from the tail, absorb the blood with filter paper every 30 s until the blood no longer oozes, and record the bleeding time ( Figure 4 A); in addition, after anesthesia with isoflurane, collect blood from the eyeballs, and measure the prothrombin time and activated partial thromboplastin time ( Figure 4 B, C). The measurement method is shown in the instruction manual of the kit (Regen Biotech).

[0190] 4. Activity data of compound ZYSO20

[0191] (1) Test of PLE enzymatic hydrolysis of ZYSO series compounds

[0192] Taking the release of the original drug 7-diethylamino-4-methylcoumarin by ZYSO20 as an example. Add ZYSO20 solution (100 μL, 500 μM), PBS buffer solution (4900 μL, pH = 7.4), and PLE (1 mg) to a 5 mL sample bottle. Seal it with a sealing film and incubate it in a 37 °C water bath. At regular intervals, take 200 μL of the solution, filter it through a 0.22 μm filter membrane, and analyze the components in the above solution using a suitable HPLC mobile phase and elution (Chromatographic column: Agilent, 4.6 mm × 150 mm; Mobile phase: ACN acetonitrile (A) and 0.01% HCOOH formic acid / water H2O (B); Flow rate: 1.0 mL / min; Injection volume: 5 μL. Linear gradient elution: 0 - 15 min, 10% - 90% A.). Calculate the Psoralen released at each time point according to the standard curve of 7-diethylamino-4-methylcoumarin, and calculate its release t 1 / 2 was greater than 120 h.

[0193] (2) C57BL / 6JNifdc male mice were selected and randomly divided into 3 groups. They were injected via the tail vein into a mouse model with middle cerebral artery occlusion (MCAO). The dose was 100 μmol / kg. The mice were sacrificed by cervical dislocation at 30 min, 1 h, and 2 h respectively. Organs such as the mouse brain, liver, kidney, heart, and spleen were taken, and fluorescence monitoring was immediately carried out on them using an in vivo imaging system for animals (Tanon, ABLX6). The results showed that at 30 min, 1 h, and 2 h, the ZYSO20 group, which was originally non-fluorescent, showed fluorescence compared with the blank group, and showed stronger fluorescence signals in multiple organs compared with the original drug group, indicating that the strategy of fluorescence recovery caused by the release of hydrogen sulfide is indeed somewhat feasible. Figure 5 )

Claims

1. A hydrogen sulfide prodrug of a coumarin compound represented by formula (I) or a pharmaceutically acceptable salt thereof, Wherein: R1 and R2 are each independently selected from H, hydroxy, amino, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, or R1 and R2 together with the adjacent carbon atom form R3, R4, R5, and R6 are each independently selected from H, hydroxy, amino, nitro, cyano, Or R3 and R4 form an unsubstituted or R-substituted carbon atom with its adjacent carbon atom. A The substituted aromatic ring is either unsubstituted or R B or R4 and R5 form an unsubstituted or substituted heteroaromatic ring with its adjacent carbon atom. A The substituted aryl group may be unsubstituted or R B or R5 and R6 form an unsubstituted or substituted heteroaromatic ring with its adjacent carbon atom. A The substituted aryl group may be unsubstituted or R B Substituted heteroaromatic ring; the substituent R A , R B Each is independently selected from H, hydroxyl, amino, nitro, cyano, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl; m is 0, 1 or 2; preferably, m is 0 or 1; R a and R b are independently selected from hydroxy, C1-C6 alkyl; preferably, R a and R b are independently selected from hydroxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

2. The hydrogen sulfide prodrug or a pharmaceutically acceptable salt thereof according to claim 1, wherein R1 is H; preferably, R2 is selected from H, hydroxyl, amino, nitro, cyano, hydroxylamine, C1-C4 alkyl, C1-C4 haloalkyl; more preferably, R2 is selected from H, hydroxyl, amino, nitro, cyano, methyl, ethyl, n-propyl, isopropyl, C1-C3 haloalkyl, 3. The hydrogen sulfide prodrug or a pharmaceutically acceptable salt thereof according to claim 1, wherein R3 is selected from H; preferably, R4 is selected from H, hydroxyl, amino, nitro, cyano; preferably, R5 is selected from H, hydroxyl, amino, nitro, cyano, Preferably, R6 is selected from H.

4. The hydrogen sulfide prodrug or a pharmaceutically acceptable salt thereof according to claim 1, wherein The aromatic ring is a benzene ring or a naphthalene ring, preferably a benzene ring; more preferably, R A is H, hydroxyl, amino, nitro, methyl, ethyl, methoxy, ethoxy, trifluoromethyl; preferably, the heteroaromatic ring is a 5- to 6-membered heteroaromatic ring having 1 to 3 members selected from N, O or S, preferably, the heteroaromatic ring is selected from a furan ring, a thiophene ring, a pyrrole ring, a pyrazole ring, an imidazole ring, an oxazole ring, a thiazole ring, a pyridine ring, a pyrimidine ring; more preferably, R B is preferably H, hydroxyl, amino, nitro, methyl, ethyl, methoxy, ethoxy, trifluoromethyl; more preferably, R B is H.

5. A hydrogen sulfide prodrug represented by formula (II) or a pharmaceutically acceptable salt thereof, Among them, R is selected from a hydroxyl group or a C1-C6 alkyl group; preferably, R is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

6. A hydrogen sulfide prodrug of a coumarin compound represented by the following specific structure or a pharmaceutically acceptable salt thereof:

7. The hydrogen sulfide prodrug according to any one of claims 1 to 6 can release hydrogen sulfide and the original drug under the action of an enzyme in vivo.

8. A preparation method of the hydrogen sulfide prodrug according to claim 1 or claim 5: A preparation method of the coumarin compound represented by formula (I): Among them, R1, R2, R3, R4, R5, R6 are as described in claim 1; A preparation method of the hydrogen sulfide prodrug of the compound represented by formula (II): wherein, R is as described in claim 5.

9. A pharmaceutical composition comprising the hydrogen sulfide prodrug according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

10. Use of the hydrogen sulfide prodrug according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9 in the preparation of a drug for treating stroke; preferably in the preparation of a drug for treating ischemic stroke.

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