Drug design strategy for releasing hydrogen sulfide based on enzymolysis of thiocarbonyl and application of drug design strategy in indole parent nucleus drug modification

Abstract

The invention discloses a drug design strategy for releasing hydrogen sulfide based on enzymolysis of thiocarbonyl and application of the drug design strategy in indole parent nucleus drug modification, a hydrogen sulfide prodrug is characterized in that an existing drug with a carboxylic acid structure is vulcanized to form a thiocarboxylic acid structure, and the thiocarboxylic acid structure can release H2S under an enzyme catalysis condition; thiocarboxylic acid can be hydrolyzed by esterase and released to obtain hydrogen sulfide, toxic and side effects of existing drugs are improved by means of anti-inflammation, myocardial protection, gastrointestinal tract protection and the like of hydrogen sulfide, physiological functions of hydrogen sulfide are exerted at the same time, and clinical application of hydrogen sulfide is achieved.

Description

[0001] This application is a divisional application of Chinese invention patent No. 202311051861.8, filed on August 21, 2023, entitled "Prodrug Design Strategy Based on Direct Esterase Hydrolysis of Thiocarboxylic Acids to Release Hydrogen Sulfide and Its Application in the Modification of Carboxylic Acid Drugs". Technical Field

[0002] This invention relates to the field of pharmaceutical chemistry, specifically to a drug design strategy based on the release of hydrogen sulfide through enzymatic hydrolysis of thiocarbonyl groups and its application in the modification of indole nuclei drugs. Background Technology

[0003] Hydrogen sulfide (H2S) was previously considered a toxic gas with a rotten egg smell, but recent studies have found that H2S is the third gaseous signaling molecule after CO and NO, participating in various physiological and pathological processes. Research indicates that H2S can dilate blood vessels and lower blood pressure, reduce tissue damage by resisting oxidative stress, regulate gut microbiota, and protect mucous membranes. However, high concentrations of H2S are toxic, and to fully utilize its properties as a gaseous signaling molecule, its release must be controlled and slow; therefore, the development of H2S prodrugs is necessary.

[0004] Currently, various hydrogen sulfide prodrugs have been developed, mainly including naturally hydrolyzed types: inorganic sulfide salts (Na2S, NaHS, CaS, etc.), Lawsson reagent derivatives (GYY4137), and response-triggered types.

[0005] The prodrug based on esterase-responsive trimethyl cyclization releases hydrogen sulfide. Its release mechanism is that the esterase hydrolyzes the ester bond in the structure, and the generated intermediate releases hydrogen sulfide and lactone through cyclization reaction. However, the structure of the esterase-responsive hydrogen sulfide prodrug is too complex, and its pharmacokinetic and pharmacological properties in vivo are unknown, which is not conducive to its clinical application.

[0006] The release mechanism of COS-based hydrogen sulfide prodrugs involves releasing COS in response to signals such as ROS, esterases, pH, thiols in vivo, and light in vitro. COS is further hydrolyzed by carbonic anhydrase to release hydrogen sulfide and CO2. However, COS itself is cytotoxic and has limited prospects for biological applications.

[0007] Hydrogen sulfide prodrugs based on thioester bonds release through the attack of thiols on the thioester bond in vivo, releasing sulfides. The sulfides then undergo a sulfur exchange reaction with the thiols to further release hydrogen sulfide. The metabolic pathways of thiol-responsive hydrogen sulfide prodrugs differ between in vivo and in vitro. Furthermore, under high oxidative stress conditions in vivo, thiols are often oxidized to disulfides, which is detrimental to drug release.

[0008] The pH-sensitive H2S prodrug of thiophosphate involves the nucleophilic carboxylic acid attacking the phosphate center under acidic conditions to form a stable five-membered ring, accompanied by the release of H2S. The complex structure of this compound hinders its application in pharmaceuticals, and its in vivo metabolic pathway is unknown.

[0009] The release mechanism of geminal dithiol hydrogen sulfide prodrug involves protecting the unstable geminal dithiol with a photosensitive group (2-nitrobenzyl) or a carbonyl group. Under ultraviolet light irradiation or when attacked by thiols, the geminal dithiol is released, followed by hydrolysis to release H2S. It requires stimulation by thiols in vivo to release hydrogen sulfide, which means that its effect may be limited by tissues and organs rich in thiols. Under high oxidative stress, thiols may be oxidized to form disulfide, which is not conducive to drug release.

[0010] The release mechanism of the thioamino acid hydrogen sulfide prodrug involves the reaction of thioamino acid with bicarbonate to form a carbamate intermediate, followed by a cyclization reaction to generate N-carboxylic anhydride with the release of H2S. The in vivo drug metabolism may not be consistent with the hypothesis.

[0011] Arylthioamide prodrugs are reported to have mechanisms of slow, spontaneous hydrolysis or accelerated hydrolysis by high concentrations of thiols, releasing hydrogen sulfide. However, they lack organ- or tissue-specific release pathways, exhibit uncertainties in efficacy and dosage, and may produce adverse reactions at high doses. Furthermore, their complex structures make them difficult to integrate with drugs.

[0012] It also includes natural polysulfides with unclear release mechanisms and noranisole (ADT-OH), etc.

[0013] Due to the aforementioned problems with hydrogen sulfide prodrugs, none are currently used clinically. Furthermore, many drugs used clinically suffer from poor efficacy or severe toxic side effects. For example, naturally hydrolyzable prodrugs hydrolyze to generate hydrogen sulfide in aqueous environments, resulting in uncontrollable and unpredictable release, thus limiting their application. Other responsive release prodrugs have overly complex chemical structures, making it difficult to accurately determine whether the efficacy is due to the prodrug carrier or the H2S itself, and their pharmacokinetics and pharmacological properties are unknown. Moreover, for thiol-sensitive H2S prodrugs, research indicates that during ischemia-reperfusion injury or inflammation, the internal environment is in a highly oxidized state, and free thiols are often in the disulfide form, making it difficult to truly achieve thiol-based prodrug activation. The cardiovascular drug butylphthalide, due to its slow onset of action, often requires combination therapy with other drugs in clinical use. Long-term use of nonsteroidal anti-inflammatory drugs such as diclofenac sodium and aspirin can lead to severe gastrointestinal, hepatic, and renal toxicity.

[0014] In summary, the drug-like properties of hydrogen sulfide prodrugs reported in current research are all poor, making it difficult to achieve clinical use. Summary of the Invention

[0015] In view of the problems of existing drugs, the present invention aims to prepare a more stable and effective hydrogen sulfide prodrug.

[0016] In a first aspect, the hydrogen sulfide prodrug provided by the present invention is to sulfide an existing drug with a carboxylic acid structure to form a thiocarboxylic acid structure, which can release H2S under enzyme catalysis.

[0017] Secondly, the present invention also provides a method for forming H2S, wherein the hydrogen sulfide prodrug provided by the present invention is contacted with an enzyme, and the thiocarboxylic acid structure is formed into a carboxylic acid structure under enzyme catalysis and H2S is released.

[0018] The enzymes described in this invention can include any enzyme capable of breaking down thiocarboxylic acid structures into carboxylic acid structures and releasing H2S. Through various in vivo and in vitro experiments, the inventors have discovered that multiple enzymes in vivo can achieve this function. For example, pharmacokinetic studies showed that carboxylic acid formation could be observed when the drug was injected into the tail vein as a prodrug of this invention containing a thiocarboxylic acid structure. This patent embodiment uses porcine liver esterase (PLE) as an example, but is not limited to this enzyme.

[0019] This invention is the first to demonstrate that compounds with a thiocarboxylic acid structure can be hydrolyzed by enzymes to release H2S under enzymatic catalysis. Furthermore, by using thiocarboxylic acid as a novel carrier for hydrogen sulfide, the hydrolysis rate of thiocarboxylic acid can be controlled by esterases, thereby slowly releasing hydrogen sulfide. Compared to some existing naturally hydrolyzed precursors that can hydrolyze to generate hydrogen sulfide in an aqueous environment, the compound described in this invention remains stable in an aqueous environment without enzymes and can only slowly release H2S through enzymatic catalysis. This achieves a slow and controllable release of H2S during treatment, avoiding the uncontrollable risks associated with the localized rapid release of hydrogen sulfide donors during use. This discovery has significant implications for the biomedical field.

[0020] Specifically, the thiocarboxylic acid structure in the prodrug of the present invention releases hydrogen sulfide in the following manner:

[0021] .

[0022] Thirdly, the present invention provides a series of hydrogen sulfide prodrugs to demonstrate the aforementioned points of the present invention.

[0023] This invention is the first to use thiocarboxylic acids in the preparation of hydrogen sulfide prodrugs. The inventors discovered that drugs with a carboxylic acid structure in their active pharmaceutical ingredients can all undergo sulfidation of the carboxylic acid structure to form the thiocarboxylic acid structure of this invention. In some embodiments, the hydrogen sulfide prodrug provided by this invention, wherein the drug before sulfidation can be a drug for treating the following diseases: drugs for treating cardiovascular and cerebrovascular diseases, anti-inflammatory drugs, antipyretic analgesics, drugs for rheumatoid arthritis, anti-gout drugs, drugs for lowering blood pressure, lowering blood lipids, or reducing hepatotoxicity caused by drug-induced GSH consumption or mitochondrial damage; in some examples, such as drugs with a -COOH structure, including: clofibrate lipid-lowering drugs, angiotensin II receptor blockers (ARBs), sulfonamides, indole drugs, nonsteroidal anti-inflammatory drugs (NSAIDs), etc. In some more specific examples, these drugs include, but are not limited to, diclofenac, ketoprofen, indolepropionic acid, indomethacin, telmisartan, gemfibrozil, naproxen, oxaprazin, ibuprofen, flurbiprofen, probenecid, etc. Other drugs with carboxylic acid structures can also have their carboxylic acid structures sulfidated to form the precursor compounds of this invention, all of which can achieve the effect of releasing hydrogen sulfide.

[0024] In some instances, the hydrogen sulfide prodrug of the present invention is a sulfidation of an existing sartan drug with a -COOH structure to form a thiocarboxylic acid structure, which can release H2S under enzyme catalysis. The sartan drug is any sartan drug with a -COOH structure used clinically, such as valsartan, eprosartan, and telmisartan. When multiple -COOH structures are present, any one or more -COOH structures can be sulfided to form a thiocarboxylic acid structure.

[0025] In other instances, the hydrogen sulfide prodrug of the present invention has the structure shown in formula (III):

[0026]

[0027] In compound (III): Z + Selected from Na + K + H + ;

[0028] L2 is selected from covalent bonds, -(CH2). n - or C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4;

[0029] R7 is selected from H and C1-C6 alkyl groups;

[0030] R8 is selected from H, -C(=O)-R 10C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; R 10 Selected from substituted or unsubstituted C6-C 10 The aryl or substituted or unsubstituted C3-C10 heteroaryl group, wherein the substituent is selected from H, halogen, -OH, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, and C1-C4 haloalkyl;

[0031] R9 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy.

[0032] In some specific schemes, L2 is selected from -(CH2). n -; n is selected from 0, 1, 2 or 3.

[0033] In some specific schemes, R7 is selected from H and C1-C4 alkyl groups; in some specific examples, R7 is selected from methyl, ethyl, propyl, isopropyl, butyl, and tert-butyl groups.

[0034] In some specific schemes, R8 is selected from H or -C(=O)-R 10 ;R 10 The following groups, whether substituted or unsubstituted, are selected: phenyl, naphthyl, anthraquinyl, oxazolyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolinyl; the substituents are selected from H, halogen, -OH, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

[0035] In some specific schemes, R9 is selected from H, halogen, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy or C1-C4 haloalkyl.

[0036] In some embodiments, the present invention provides the following specific hydrogen sulfide prodrugs:

[0037] .

[0038] The hydrogen sulfide prodrug described in this invention can be synthesized by sulfidation of the carboxylic acid of existing drugs using commonly used chemical synthesis methods in the prior art.

[0039] The present invention also provides a pharmaceutical composition comprising the hydrogen sulfide prodrug described herein and a pharmaceutically acceptable carrier.

[0040] The hydrogen sulfide prodrug described in this invention can be administered to the subject, either the compound of this invention or a pharmaceutical composition containing the composition of this invention, in an environment sufficient to form hydrogen sulfide. The inventors have experimentally discovered that hydrogen sulfide release can be achieved via oral, intravenous, and other routes.

[0041] The present invention also provides the use of the aforementioned hydrogen sulfide prodrug in the preparation of medicaments for treating cardiovascular and cerebrovascular diseases, anti-inflammatory, antipyretic and analgesic, rheumatoid arthritis, gout, lowering blood pressure, lowering blood lipids, or reducing hepatotoxicity caused by drug-induced GSH consumption or mitochondrial damage.

[0042] The compounds of the present invention can be used alone as a single therapeutic agent or in combination with other active pharmaceutical ingredients.

[0043] The hydrogen sulfide prodrug described in this invention has the following advantages over the prior art:

[0044] (1) Different physiological functions of hydrogen sulfide can be achieved by simply changing the structure and chemical properties of carboxylic acids. In this process, it is not necessary to change the original drug skeleton, but only to make minor changes to the structure of the drug, which can greatly shorten the time and cost of drug development.

[0045] (2) The hydrogen sulfide prodrug of the present invention is derived from thiocarboxylic acids that can be hydrolyzed by esterases to release hydrogen sulfide. The anti-inflammatory, cardioprotective, and gastrointestinal protective properties of hydrogen sulfide are utilized to improve the toxic side effects of existing drugs, while simultaneously leveraging the physiological functions of hydrogen sulfide itself to achieve its clinical application. Based on this characteristic, the hydrogen sulfide prodrug of the present invention has the advantage of improving efficacy while reducing the toxic side effects of drugs. For example, in the treatment of cerebral infarction and inflammation, the use of the thiocarboxylic acid compounds synthesized in this invention can enhance the efficacy of the drug while reducing its adverse effects on human health.

[0046] Unless otherwise specified, the terms used in this invention have the following definitions:

[0047] "alkyl" refers to a saturated aliphatic hydrocarbon group with 1-30 carbon atoms, including straight-chain and branched groups (the numerical range mentioned in this application, such as "1-20", refers to the group, which is an alkyl group and can contain 1, 2, 3 carbon atoms, etc., up to 20 carbon atoms). Examples include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, pentyl, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, pentyl, neopentyl, hexyl, heptyl, octyl, etc.

[0048] "Halogenated alkyl" refers to a halogen-substituted alkyl group, which is replaced by one or more identical or different halogen atoms, such as -CH2Cl, -CF3, -CH2CF3, -CH2CCl3, etc.

[0049] "Alkoxy" indicates -O- (unsubstituted alkyl) or -O- (unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexyloxy.

[0050] “Cycloalkyl” means a monocyclic or fused ring group consisting entirely of carbon (“fused” rings mean that each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system), wherein one or more rings do not have a fully connected π-electron system. Examples of cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cyclohepttriene.

[0051] "Aryl" refers to an all-carbon monocyclic or fused polycyclic group with 1 to 12 carbon atoms and a fully conjugated π-electron system. Non-limiting examples of aryl groups include phenyl, naphthyl, and anthracene. Aryl groups can be substituted or unsubstituted. When substituted, the substituents are preferably one or more, more preferably one, two, or three, and even more preferably one or two.

[0052] "Heteroaryl" refers to a stable monocyclic ring with up to 3-10 atoms or a bicyclic carbon ring with up to 3-10 atoms per ring, wherein at least one ring is an aromatic ring and contains 1-4 heteroatoms selected from O, N, and S. Non-limiting examples of unsubstituted heteroaryl groups include pyrroleyl, furanyl, thiopheneyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, purinyl, acridineyl, carbazoleyl, cenylyl, quinoxolinyl, indolyl, benzotriazolyl, benzothiopheneyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl, and carbazoleyl. Heteroaryl groups can be substituted or unsubstituted. When substituted, the substituents are preferably one or more, more preferably one, two, or three, and even more preferably one or two.

[0053] "Spirocyclic": When a molecule has two rings that share a carbon atom, the system is called a spirocyclic.

[0054] "Bridge ring" refers to a polycyclic hydrocarbon that shares two or more carbon atoms (bridgehead carbons). Based on the number of rings, it is classified into bicyclic hydrocarbons, tricyclic hydrocarbons, tetracyclic hydrocarbons, etc.

[0055] "Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine. Attached Figure Description

[0056] Figure 1 The evaluation of the efficacy of ZYSA01 in cerebral infarction is as follows: A represents the infarct area; B represents the quantitative analysis of the infarct area; C represents the serum LDH level analysis; and D represents the tissue MDA level analysis.

[0057] Figure 2 A represents the activity data of ZYSH06; B represents the infarct area; C represents the quantitative analysis of the infarct area; D represents the serum LDH level analysis; and D represents the tissue MDA level analysis.

[0058] Figure 3 The plasma concentration-time curves of ZYSH21 and DCF interconversion after oral administration (10 mg / kg) are shown.

[0059] Figure 4 The graph shows the changes in paw swelling rate in rats in the DCF and ZYSH21 groups after administration of 10 mg / kg by gavage, n=8.

[0060] Figure 5 The percentage of gastric ulcers in rats in the DCF and ZYSH21 groups after administration of 10 mg / kg by gavage, n=8.

[0061] Figure 6 Serum AST and ALT levels 24 h after administration of DCF (150 mg / kg, ip) and ZYSH21 (157 mg / kg, ip).

[0062] Figure 7 Serum uric acid levels in rats in the DCF and ZYSH21 groups were measured after administration of 10 mg / kg by gavage (n=8).

[0063] Figure 8 The plasma concentration-time curves of ZYSH31 and NPX interconversion after oral administration (10 mg / kg) are shown.

[0064] Figure 9 The graph shows the change in paw swelling rate in rats in the NPX and ZYSH31 groups after administration of 10 mg / kg by gavage, n=6.

[0065] Figure 10 The percentage of gastric ulcers in rats in the NPX and ZYSH31 groups after administration of 10 mg / kg by gavage, n=6.

[0066] Figure 11 Serum AST and ALT levels 24 h after administration of NPX (150 mg / kg, ip) and ZYSH31 (159 mg / kg, ip), n=6.

[0067] Figure 12 Serum albumin levels in rats in the NPX and ZYSH31 groups were measured after administration of 10 mg / kg by gavage (n=6). Detailed Implementation

[0068] The following examples are provided to better understand the present invention, but are not intended to limit the invention. Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were purchased from conventional biochemical reagent stores.

[0069] Example 1 Compound ZYSA01

[0070]

[0071] Thiobenzoic acid (purchased from a reagent supplier) (1.00 g, 7.24 mmol, 0.85 mL) was placed in a round-bottom flask equipped with a magnetic stirrer, dissolved in an appropriate amount of methanol, and cooled in an ice-salt bath. After cooling, a methanol solution of NaOH (0.23 g, 5.80 mmol, 0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After the reaction solution was evaporated to dryness, anhydrous diethyl ether was added, and a white solid precipitated out. The solid was then filtered to obtain sodium thiobenzoate (1.15 g, 99%).

[0072] 1 H NMR (300 MHz, CD3OD) δ 8.15 – 7.70 (m, 2H), 7.47 – 6.94 (m, 3H).

[0073] 13 C NMR (75 MHz, CD3OD) δ 213.44, 144.40, 130.00, 128.90, 127.90,127.40, 126.96.

[0074] Example 2 Compound ZYSA02

[0075]

[0076] Following the synthesis method of target ZYSA01, crude thiophenylacetic acid was synthesized from phenylacetic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium thiophenylacetate, a white solid.

[0077] 1 H NMR (300 MHz, DMSO-d6) δ 7.08-7.19 (m, 5H), 3.68 (s, 2H).

[0078] 13 C NMR (101 MHz, CD3OD) δ 137.89, 128.85, 127.64, 125.64, 57.34.

[0079] Example 3 Compound ZYSA03

[0080]

[0081] Following the synthesis method of target ZYSA01, crude thiophenylpropionic acid was synthesized from phenylpropionic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium thiophenylpropionic acid, a white solid.

[0082] 1 H NMR (500 MHz, DMSO-d6) δ 7.22 (t, J = 10.0 Hz, 2H), 7.10-7.17 (m,3H), 2.79 (t, J = 5.0 Hz, 2H), 2.63 (t, J = 10.0 Hz, 2H).

[0083] 13 C NMR (75 MHz, DMSO-d6) δ 215.98, 143.31, 128.69, 128.55, 125.76, 53.24, 33.75.

[0084] Example 4 Compound ZYSA04

[0085]

[0086] 4-Methylbenzoic acid (1.00 g, 7.34 mmol) and Lawson's reagent (1.63 g, 4.03 mmol, 0.55 eq) were added to a two-necked round-bottom flask equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (30 mL) was added as a solvent using a syringe. The mixture was stirred at 130 °C for 30 min. The system changed from turbid to clear. The reaction was monitored by TLC and found to be almost complete. The toluene was removed by concentration under reduced pressure. The residue was dissolved in DCM and separated by silica gel column chromatography (mobile phase: pure DCM) to obtain 0.89 g of crude product.

[0087] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, dissolved in an appropriate amount of methanol, and cooled in an ice-salt bath. After cooling, a methanol solution of NaOH (0.19 g, 4.70 mmol, 0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After the reaction solution was evaporated to dryness, anhydrous diethyl ether was added, and a white solid precipitated out. After filtration, sodium 4-methylthiobenzoate (770 mg, 60%) was obtained.

[0088] 1 H NMR (500 MHz, DMSO-d6) δ 7.98 (d, J = 10.0 Hz, 2H), 7.05 (d, J = 5.0Hz, 2H), 2.30 (s, 3H).

[0089] 13C NMR (126 MHz, DMSO-d6) δ 207.96, 143.21, 138.62, 128.47, 127.74,21.32.

[0090] Example 5 Compound ZYSA05

[0091]

[0092] Following the synthesis method of ZYSA04, crude 4-methoxythiobenzoic acid was synthesized from 4-methoxybenzoic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium 4-methoxythiobenzoate, a white solid.

[0093] 1 H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J = 10.0 Hz, 2H), 6.77 (d, J =10.0 Hz, 2H), 3.75 (s, 3H).

[0094] 13 C NMR (126 MHz, DMSO-d6) δ 207.50, 160.76, 138.57, 130.16, 112.25,55.57.

[0095] Example 6 Compound ZYSA06

[0096]

[0097] p-Tolueneacetic acid and Lawson's reagent (0.55 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stirrer. A reflux flask was then placed in place. After replacing the nitrogen with nitrogen, the mixture was heated to 110°C and refluxed. The reaction was monitored by TLC. After approximately 1.5 hours of reflux, the system changed from turbid to clear, indicating that the reaction was largely complete. The reaction solution was removed, toluene was removed under vacuum, and the residue was dissolved in DCM. The solution was then loaded onto a silica gel column, and eluted with DCM to obtain the crude product (a yellow oily substance).

[0098] The crude product obtained in the previous step was placed in a round-bottom flask with a magnetic stirrer. After dissolving it with an appropriate amount of MeOH, it was placed in an ice-salt bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After removing the MeOH under vacuum, the residue was dissolved in a small amount of DCM. Anhydrous diethyl ether was added, and a white solid precipitated out. After filtration, sodium thio-p-methylphenylacetate was obtained.

[0099] 1H NMR (300 MHz, DMSO-d6) δ 7.07 (d, J = 8.0 Hz, 2H), 6.99 (d, J = 7.8Hz, 2H), 3.64 (s, 2H), 2.23 (s, 3H).

[0100] 13 C NMR (75 MHz, DMSO) δ 212.98, 136.06, 133.90, 128.99, 128.09, 58.04, 20.70.

[0101] Example 7 Compound ZYSA07

[0102]

[0103] Following the synthesis method of target ZYSA01, crude 4-chlorothiophenylpropionic acid was synthesized from 4-chlorophenylpropionic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium 4-chlorothiophenylpropionic acid, a white solid.

[0104] 1 H NMR (300 MHz, CD3OD) δ 7.17-7.23 (m, 4H), 2.83-2.95 (m, 4H).

[0105] 13 C NMR (101 MHz, CD3OD) δ 140.75, 131.00, 129.63, 127.84, 52.23,32.80.

[0106] Example 8 Compound ZYSA08

[0107]

[0108] Following the synthesis method of target ZYSA01, crude 4-methoxythiophenylpropionic acid was synthesized from 4-methoxyphenylpropionic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium 4-methoxythiophenylpropionic acid, a white solid.

[0109] 1 H NMR (300 MHz, CD3OD) δ 7.07 (d, J = 9.0 Hz, 2H), 6.77-6.80 (m, 2H), 3.70 (s, 3H), 2.69-2.74 (m, 2H), 2.55-2.59 (m, 2H).

[0110] 13C NMR (101 MHz, CD3OD) δ 157.84, 133.94, 128.83, 128.77, 113.28, 54.24, 52.99, 32.76.

[0111] Example 9 Compound ZYSB01

[0112]

[0113] γ-Butyrolactone (2 g, 23.2 mmol) and Lawson's reagent (5.6 g, 13.9 mmol, 0.6 eq) were added to a two-necked round-bottom flask equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (30 mL) was added as a solvent using a syringe. The mixture was stirred at 130 °C for 5 h. The system changed from turbid to clear. The reaction was monitored by TLC and found to be almost complete. The toluene was removed by concentration under reduced pressure. The mixture was then separated by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 5 / 1) to obtain thio-γ-butyrolactone, a yellow oily liquid (647 mg, 30%).

[0114] Thio-γ-butyrolactone (193 mg, 1.9 mmol) was added to a single-necked round-bottom flask equipped with a magnetic stirrer and dissolved in 10 mL of methanol. Sodium hydroxide (91 mg, 2.3 mmol, 1.2 eq) was weighed, dissolved in 3 mL of water, and added to the methanol solution of thio-γ-butyrolactone. The mixture was stirred at room temperature for 3 h, and the reaction was monitored by TLC until complete. The solvent was removed by concentration under reduced pressure to obtain sodium γ-hydroxythiobutyrate, a white solid (183 mg, 70%).

[0115] 1 H NMR (300 MHz, CD3OD) δ 3.57 (t, J = 6.0 Hz, 2H), 2.71 (t, J = 6.0Hz, 2H), 1.82-1.91 (m, 2H) . 13 C NMR (75 MHz, CD3OD) δ 221.88, 61.32, 34.48,30.28.

[0116] Example 10 Compound ZYSB02

[0117]

[0118] Following the synthesis method of ZYSB01, thio-α-methyl-γ-butyrolactone was synthesized from α-methyl-γ-butyrolactone and Lawson's reagent. Subsequently, it was alkaline hydrolyzed to obtain sodium α-methyl-γ-hydroxythiobutyrate, a white solid.

[0119] 1 H NMR (300 MHz, CD3OD) δ 3.54-3.60 (m, 2H), 2.85-2.96 (m, 1H), 1.85-1.98 (m, 1H), 1.49-1.58 (m, 1H), 1.11 (d, J = 6.0 Hz, 3H).

[0120] 13 C NMR (75 MHz, , CD3OD) δ 226.19, 60.17, 50.79, 37.99, 18.63.

[0121] Example 11 Compound ZYSB03

[0122]

[0123] Octadectic acid and Lawson's reagent (0.55 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing with nitrogen, dry THF was added as a solvent using a syringe. The reaction was carried out with stirring at 65°C, and the reaction was monitored by TLC. After about overnight, the system changed from turbid to clear. The reaction solution was removed, and THF was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (white solid).

[0124] The crude product obtained in the previous step was placed in a round-bottom flask with a magnetic stirrer. After dissolving it with an appropriate amount of DCM, it was placed in an ice-salt bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After removing the MeOH under vacuum, the residue was dissolved with a small amount of DCM. Anhydrous diethyl ether was added, and a white solid precipitated out. After filtration, sodium thiooctadecanoate was obtained.

[0125] 1 H NMR (300 MHz, DMSO-d6) δ 2.30 (t, J = 7.5 Hz, 2H), 1.43 (t, J = 7.2Hz, 2H), 1.23 (s, 28H), 0.91 – 0.79 (m, 3H).

[0126] Example 12 Compound ZYSC01

[0127]

[0128] Following the synthesis method of target ZYSA01, crude thionaphthoic acid was synthesized from naphthoic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium thionaphthoic acid, a pale yellow solid.

[0129] 1 H NMR (300 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.26 (d, J = 9.0 Hz, 1H), 7.84-7.96 (m, 2H), 7.74 (d, J = 9.0 Hz, 1H), 7.46-7.48 (m, 2H).

[0130] 13 C NMR (101 MHz, CD3OD) δ 213.07, 141.61, 134.56, 132.63, 128.91,128.00, 127.11, 126.56, 126.27, 125.67, 125.37.

[0131] Example 13 Compound ZYSC02

[0132]

[0133] Following the synthesis method of target ZYSA01, crude thionaphthaleneacetic acid was synthesized from naphthaleneacetic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium thionaphthaleneacetate, a pale yellow solid.

[0134] 1 H NMR (300 MHz, DMSO-d6) δ 7.78-7.83 (m, 2H), 7.73(d, J = 9.0 Hz,1H), 7.62 (s, 1H), 7.36-7.46 (m, 3H), 3.86 (s, 2H).

[0135] 13 C NMR (101 MHz, CD3OD) δ 135.52, 133.67, 132.26, 127.60, 127.20,127.11, 127.08, 127.02, 125.30, 124.76, 57.46.

[0136] Example 14 Compound ZYSD01

[0137]

[0138] Phenylacetylglycine (200 mg, 1.04 mmol, 1 eq), NHS (238 mg, 2.07 mmol, 2 eq), and EDCI HCl (298 mg, 1.55 mmol, 1.5 eq) were added to a two-necked round-bottom flask equipped with a magnetic stirrer. After replacing the N2 concentration, anhydrous THF (5 mL) was added using a syringe under ice bath conditions. The mixture was then brought to room temperature, and the reaction was monitored by TLC. After 12 hours of reaction, the reaction was approximately complete. The reaction solution was removed, concentrated under vacuum, and the residue was dissolved with ethyl acetate (30 mL). The organic phase was washed with water (3 × 10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to remove the solvent, yielding the active ester product (white solid, 265 mg, y = 88%).

[0139] The active ester product obtained in the previous step (200 mg, 0.68 mmol, 1 eq) was dissolved in THF (5 mL) and cooled in an ice bath. NaHS xH2O (68%) (137 mg, 1.37 mmol, 2 eq) was dissolved in water (less than 2 mL). The NaHS xH2O aqueous solution was slowly added dropwise to the THF solution of the active ester in an ice bath. After the addition was complete, the ice bath was removed and the mixture was stirred at room temperature. The reaction was monitored by TLC. After 1 hour of reaction, the reaction solution was removed, diluted under vacuum, and the pH was adjusted to 3 with 1M HCl. The aqueous phase was extracted with ethyl acetate (3 × 10 mL), the organic phases were combined, washed once with saturated brine, dried over anhydrous Na2SO4, and the organic phase was concentrated under vacuum. The residue was dissolved with a small amount of DCM and loaded onto a silica gel column. The column was eluted with DCM containing 1% acetic acid by volume. The eluent was collected, most of the solvent was removed under reduced pressure, and a large amount of PE was added. A white powdery solid precipitated out. Filter and wash the precipitate with PE to obtain the thiolated product (white powder solid, 38 mg, y=20%).

[0140] 1 H NMR (300 MHz, Methanol-d4) δ 7.36 – 7.21 (m, 5H), 4.10 (s, 2H), 3.59 (s, 2H).

[0141] 13 C NMR (75 MHz, MeOD) δ 175.56, 168.55, 140.18, 138.77, 128.96, 126.98, 45.64, 44.73, 40.58, 30.20, 21.41, 17.61.

[0142] Example 15 Compound ZYSD02

[0143]

[0144] Ibuprofenylglycine (400 mg, 1.52 mmol, 1 eq), NHS (350 mg, 3.04 mmol, 2 eq), and EDCI HCl (435 mg, 2.28 mmol, 1.5 eq) were added to a two-necked round-bottom flask equipped with a magnetic stirrer. After replacing the N2 concentration, anhydrous THF (8 mL) was added under an ice bath using a syringe. The mixture was then brought to room temperature, and the reaction was monitored by TLC. After 12 hours, the reaction was approximately complete. The reaction solution was removed, concentrated under vacuum, and the residue was dissolved with ethyl acetate (50 mL). The organic phase was washed with water (3 × 20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to remove the solvent, yielding the active ester product (white solid, 460 mg, y = 84%).

[0145] Take 300 mg of the active ester product obtained in the previous step and place it in a round-bottom flask equipped with a magnetic stirrer. Add 5 mL of THF to dissolve it and stir under ice bath. Dissolve NaHS xH2O (137 mg, 1.66 mmol, 2 eq) in water (less than 1 mL) and slowly add it dropwise to the THF solution of the active ester under ice bath. Monitor the reaction by TLC. After 5 minutes of reaction, remove the reaction solution, concentrate it under vacuum, dilute it with water, and adjust the pH to 3 with 1M HCl. Extract the aqueous phase three times with ethyl acetate (3 × 20 mL), combine the organic phases, wash once with saturated brine, dry with anhydrous sodium sulfate, combine the organic phases, concentrate them under vacuum to remove the solvent, and load the sample onto a silica gel column with a small amount of residual DCM solvent. Elute with DCM containing 1% acetic acid, collect the eluent, remove most of the solvent under reduced pressure, add a large amount of PE, and a white powdery solid precipitates. Filter, wash the precipitate with PE again, and obtain ZYSD02 (white powdery solid, 49 mg, y = 21%).

[0146] 1 H NMR (300 MHz, Methanol-d4) δ 7.26 (d, J = 8.03 Hz, 2H), 7.10 (d, J= 8.10 Hz, 2H), 4.04 (d, J = 2.74 Hz, 2H), 3.69 (q, J = 7.08 Hz, 1H), 2.44(d, J = 7.18 Hz, 2H), 1.83 (hept, J = 13.54, 6.76 Hz, 1H), 1.46 (d, J = 7.14Hz, 3H), 0.89 (d, J = 6.61 Hz, 6H).

[0147] 13C NMR (75 MHz, Methanol-d4) δ 175.56, 168.55, 140.18, 138.77,128.96, 126.98, 45.64, 44.73, 40.58, 30.20, 21.41, 17.61.

[0148] Example 16 Compound ZYSH01 (ibuprofen)

[0149]

[0150] Ibuprofen (4 mmol, 825 mg, 1 eq) and Lawson's reagent (2.2 mmol, 890 mg, 0.55 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (10 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 80 °C. The reaction was monitored by TLC. After about 1 hour, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0151] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was then concentrated under vacuum, and cyclohexane was added. A white solid precipitated out. The solid was filtered to obtain sodium thioibuprofen (white solid, 658 mg, y=67%).

[0152] 1 H NMR (400 MHz, Methanol-d4) δ 7.30 – 7.25 (m, 2H), 7.07 – 6.97 (m,2H), 4.16 (q, J = 7.1 Hz, 1H), 2.41 (d, J = 7.1 Hz, 2H), 1.81 (dp, J = 13.5,6.8 Hz, 1H), 1.40 (d, J = 7.1 Hz, 3H), 0.88 (d, J = 6.6 Hz, 6H).

[0153] 13 C NMR (101 MHz, Methanol-d4) δ 142.27, 140.36, 129.66, 128.36, 60.80, 46.11, 31.50, 22.73, 20.42.

[0154] Example 17 Compound ZYSH02 (Indomethacin)

[0155]

[0156] Indomethacin (893 mg, 3 mmol, 1 eq) was dissolved in DCM (15 mL), and EDCI HCl (690 mg, 3.6 mmol, 1.2 eq) and NHS (518 mg, 4.5 mmol, 1.5 eq) were added sequentially. The reaction was monitored by TLC. After stirring at room temperature for 2 h, the reaction was roughly complete. The reaction solution was concentrated under vacuum, and the residue was dissolved with a small amount of DCM before being loaded onto a silica gel column. Eluting with DCM yielded the active indomethacin ester (white solid, 1.24 g, y=91%).

[0157] Indomethacin active ester (1.24 g) was dissolved in THF (15 mL) and cooled in an ice bath. NaHS xH2O (68%) (546 mg, 2.73 mmol, 2 eq) was dissolved in water (less than 5 mL). The NaHS xH2O aqueous solution was slowly added dropwise to the THF solution of indomethacin active ester in the ice bath. After the addition was complete, the ice bath was removed, and the mixture was stirred at room temperature. After confirming the completeness of the reaction by TLC, the reaction solution was concentrated under vacuum, the residue was dissolved with DCM, and the solution was loaded onto a silica gel column. Eluting with DCM yielded crude thioindomethacin (pale yellow solid).

[0158] The crude thioindomethacin product was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice bath for 5 minutes. The reaction solution was concentrated under vacuum, and a small amount of DCM was added to dissolve the residue. Anhydrous diethyl ether was then added, and a white solid precipitated out. The product was filtered to obtain sodium thioindomethacin (white solid, 854 mg, y=72%).

[0159] 1 H NMR (300 MHz, Methanol-d4) δ 7.74 – 7.67 (m, 2H), 7.58 – 7.51 (m,2H), 7.13 (d, J = 2.5 Hz, 1H), 6.95 (d, J = 8.9 Hz, 1H), 6.62 (dd, J = 9.0,2.6 Hz, 1H), 4.04 (s, 2H), 3.80 (s, 3H), 2.26 (s, 3H), 1.90 (s, 1H).

[0160] 13C NMR (101 MHz, Methanol-d4) δ 180.47, 167.57, 154.63, 140.44,134.56, 132.15, 132.08, 130.72, 129.98, 129.83, 111.51, 110.80, 108.50,101.90, 56.29, 47.65, 24.23, 11.85, -15.68.

[0161] Example 18 Compound ZYSH03 (ketoprofen)

[0162]

[0163] Ketoprofen (763 mg, 3 mmol, 1 eq) was dissolved in DCM (15 mL), and EDCI HCl (690 mg, 3.6 mmol, 1.2 eq) and NHS (518 mg, 4.5 mmol, 1.5 eq) were added sequentially. The reaction was monitored by TLC. After stirring in the dark at room temperature for 2 h, the reaction was roughly complete. The reaction solution was concentrated under vacuum, and the residue was dissolved with a small amount of DCM before being loaded onto a silica gel column. Eluting with DCM yielded the active ketoneprofen ester (white solid, 874 mg, y=83%).

[0164] Ketoprofen active ester (800 mg, 2.27 mmol, 1 eq) was dissolved in THF (15 mL) and cooled in an ice bath. NaHS xH2O (68%) (455 mg, 4.55 mmol, 2 eq) was dissolved in water (less than 5 mL). The NaHS xH2O aqueous solution was slowly added dropwise to the THF solution of ketoprofen active ester in an ice bath. After the addition was complete, the ice bath was removed, and the mixture was stirred at room temperature. After confirming the completeness of the reaction by TLC, the reaction solution was concentrated under vacuum, the residue was dissolved with DCM, and the solution was loaded onto a silica gel column. Eluting with DCM yielded the thioketoprofen product (pale yellow solid).

[0165] The crude thioprofen product was placed in a round-bottom flask equipped with a magnetic stirrer, dissolved in an appropriate amount of DCM, and cooled in an ice bath. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice bath for 5 minutes. The reaction solution was concentrated under vacuum, cyclohexane was added, and the purity was determined by TLC. Low-polarity impurities were continuously washed away with DCM / cyclohexane. The solution was dissolved in tert-butanol by rotary evaporation, and lyophilized to obtain sodium thioprofen (pale yellow foamy solid, 307 mg, y = 35%).

[0166] 1H NMR (300 MHz, Chloroform-d) δ 7.83 – 7.70 (m, 4H), 7.65 – 7.43 (m,5H), 4.25 – 3.89 (m, 1H), 1.58 (dd, J = 19.6, 7.1 Hz, 3H).

[0167] Example 19 Compound ZYSH04 (gemfibrozil)

[0168]

[0169] Gemfibrozil and Lawson's reagent (0.55 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene was added as a solvent using a syringe. The mixture was stirred at 90°C, and the reaction was monitored by TLC. After about 2 hours, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, the toluene was evaporated to dryness, and the residue was dissolved in DCM. The solution was then loaded onto a silica gel column, and the crude product (pink oily substance) was obtained by elution with DCM.

[0170] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer. An appropriate amount of MeOH was added to dissolve it. The flask was then placed in an ice-salt bath to cool it down. After cooling, a methanol solution of NaOH (0.8 eq) was added. The mixture was stirred in the ice-salt bath for 5 minutes. After the reaction solution was evaporated to dryness, anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thiogefibrazine (white solid).

[0171] 1 H NMR (400 MHz, Methanol-d4) δ 6.92 (d, J = 7.5 Hz, 1H), 6.65 (d, J =1.5 Hz, 1H), 6.59 (d, J = 7.4 Hz, 1H), 3.95 – 3.82 (m, 2H), 2.26 (s, 3H), 2.12 (s, 3H), 1.77 (d, J = 3.4 Hz, 3H), 1.21 (s, 6H).

[0172] 13 C NMR (101 MHz, Methanol-d4) δ 157.06, 136.12, 129.69, 122.99,120.14, 111.61, 68.41, 50.92, 38.81, 26.75, 25.15, 20.10, 14.64.

[0173] Example 20 Compound ZYSH05 (Probenecid)

[0174]

[0175] Probenecid (285 mg, 1 mmol, 1 eq) was added to a two-necked round-bottom flask (oven-dried) with a magnetic stir bar. After replacing the nitrogen with nitrogen, DCM (2 mL), oxaloyl chloride (170 μL, 4 mmol, 2 eq), and DMF (2 drops) were added sequentially using a syringe. The mixture was stirred at room temperature for 1 h. The reaction solution was then removed and concentrated under vacuum to obtain a yellow oily substance (310 mg), which was then dissolved in dry THF (3 mL).

[0176] The THF solution of probenecid chloride was cooled and stirred in an ice bath. Sodium hydrosulfide hydrate (136 mg, 2 mmol, 2 eq) was dissolved in a small amount of water (less than 2 mL) and slowly added dropwise to the THF solution of probenecid chloride. After the addition was complete, the reaction was stirred in an ice bath for 1 h, and then transferred to room temperature for 1 h. The reaction was quenched with 1 M HCl. The aqueous phase was extracted three times with ethyl acetate (3 × 20 mL). The combined organic layers were dried with anhydrous NaSO4 and concentrated under vacuum to obtain crude thioprobenecid (yellow oil).

[0177] The crude product obtained in the previous step was placed in a round-bottom flask with a magnetic stirrer. After dissolving it with an appropriate amount of MeOH, it was placed in an ice-salt bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After removing the MeOH under vacuum, the residue was dissolved with a small amount of DCM. Anhydrous diethyl ether was added, and a bright yellow solid precipitated out. After filtration, sodium thiopropanesulfonate (bright yellow solid, 119 mg, y = 36%) was obtained.

[0178] 1 H NMR (300 MHz, Methanol-d4) δ 8.26 – 8.18 (m, 2H), 7.77 – 7.70 (m,2H), 3.10 (d, J = 7.6 Hz, 4H), 1.55 (h, J = 7.4 Hz, 4H), 0.88 (t, J = 7.4 Hz,6H).

[0179] 13 C NMR (101 MHz, Methanol-d4) δ 212.67, 149.16, 141.93, 129.60,127.23, 51.31, 23.11, 11.43.

[0180] Example 21 Compound ZYSH06 (telmisartan)

[0181]

[0182] Telmisartan (500 mg, 0.99 mmol) was placed in a three-necked flask, and the flask was evacuated to remove nitrogen. DCM was added to dissolve the flask, and oxalyl chloride (137 mg, 125 μL) was added dropwise under ice bath conditions, followed by a catalytic amount of DMF. The mixture was then transferred to room temperature and reacted for 1 h. Dry hydrogen sulfide gas was then introduced, and the reaction was monitored by TLC. The DCM:MeOH ratio was 15:1. The mixture was then purified by column chromatography. 300 mg (57%) of a yellow solid was obtained. 1H NMR (300 MHz, Methanol-d4) δ 8.09 (s, 1H), 7.98 (d, J = 5.2 Hz,1H), 7.89 (d, J = 9.4 Hz, 2H), 7.83 – 7.65 (m, 3H), 7.57 (d, J = 6.4 Hz, 1H),7.46 (d, J = 6.8 Hz, 3H), 7.39 (d, J = 7.3 Hz, 3H), 5.92 (s, 2H), 4.04 (s,3H), 2.86 (s, 3H), 1.95 (q, J = 7.6 Hz, 2H), 1.15 (t, J = 7.4 Hz, 3H).

[0183] Example 22 Compound ZYSH21 (diclofenac)

[0184]

[0185] Synthesis of sodium 2-[(2,6-dichlorophenyl)amino]-phenylthioacetate (ZYSH21). Sodium 2-[(2,6-dichlorophenyl)amino]-phenylacetate (504 mg, 1.58 mmol) and Lawson's reagent (351 mg, 0.869 mmol) were added to a round-bottom flask and dissolved in 20 mL of dichloromethane. The reaction was carried out at room temperature for 2.5 h. TLC monitoring showed the reaction was nearly complete. The dichloromethane was removed by concentration under reduced pressure to obtain the crude product. The crude product was separated by silica gel column chromatography (mobile phase: pure DCM). The pure product was dissolved in 1.5 mL of dichloromethane and 0.4 mL of 0.1 M NaHCO3 ethanol solution, allowed to stand at -20 °C for 30 min, the solvent was removed by concentration under reduced pressure, and the product was recrystallized in diethyl ether to obtain the final product as a white solid (301 mg, 60%). 1H NMR (300MHz, DMSO-d6): δ9.17 (s, 1H), 7.46 (d, J=8.0 Hz, 2H), 7.25-7.04 (m, 2H), 6.95 (d, J=7.5Hz, 1H), 6.79 (d, J=7.3Hz, 1H), 6.26 (d, J=7.8 Hz, 1H), 3.87 (s, 2H); 13C NMR (CDCl3): 214.3, 143.4, 138.4, 130.9, 129.6, 129.5, 128.9, 126.4, 124.7, 120.8, 116.6, 56.8.

[0186] Example 23 Compound ZYSH31 (Naproxen)

[0187]

[0188] Naproxen (4 mmol, 920 mg, 1 eq) and Lawson's reagent (2.2 mmol, 890 mg, 0.55 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (10 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 80 °C. The reaction was monitored by TLC. After about 1 hour, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, the toluene was evaporated to dryness, and the residue was dissolved in DCM. The solution was loaded onto a silica gel column, and the crude product (a dark pink oily substance) was obtained by elution with DCM.

[0189] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer. An appropriate amount of DCM was added to dissolve it, and the flask was placed in an ice-salt bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in the ice-salt bath for 5 minutes. After the reaction solution was evaporated to dryness, anhydrous diethyl ether was added, and a white solid precipitated out. The product was then filtered to obtain sodium thionaproxen (white solid, 800 mg, y=74%).

[0190] 1 H NMR (500 MHz, Methanol-d4) δ 7.73 – 7.61 (m, 3H), 7.52 (dd, J =8.6, 1.7 Hz, 1H), 7.15 (d, J = 2.5 Hz, 1H), 7.06 (dd, J = 8.9, 2.5 Hz, 1H), 4.32 (q, J = 7.1 Hz, 1H), 3.88 (s,3H), 1.50 (d, J = 7.1 Hz, 3H).

[0191] 13 C NMR (126 MHz, Methanol-d4) δ 223.91, 158.64, 140.29, 134.84,130.49, 130.16, 128.06, 127.35, 126.64, 119.28, 106.57, 61.10, 55.70, 20.41.

[0192] (ESI): m / z calculated for C 14 H 13 Na O2 S requires 291.04262 for [M+Na] + , found291.04247.

[0193] Example 24 Synthesis of compound ZYSH07:

[0194]

[0195] Flurbiprofen (4 mmol, 976 mg, 1 eq) and Lawson's reagent (2.2 mmol, 890 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (20 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 95 °C. The reaction was monitored by TLC. After about 30 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a dark pink oily substance).

[0196] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The solid was then filtered to obtain sodium thioflurprofen (white solid, 555 mg, y=49%).

[0197] 1 H NMR (300 MHz, Methanol-d4) δ 7.52 (dt, J = 8.2, 1.5 Hz, 2H), 7.46– 7.31 (m, 4H), 7.29 – 7.19 (m, 2H), 4.24 (q, J = 7.1 Hz, 1H), 1.47 (d, J =7.1 Hz, 3H).

[0198] 13 CNMR(75MHz,Methanol-d4)δ222.80,162.37,159.12,146.94,137.30,131. 12,129.90,129.36,128.39,127.87,127.69,125.01,115.80,60.61,20.26.

[0199] Example 25 Synthesis of compound ZYSH08:

[0200]

[0201] Oxaprazin (3 mmol, 879 mg, 1 eq) and Lawson's reagent (1.8 mmol, 728 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (15 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 90 °C. The reaction was monitored by TLC. After about 30 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (pink oily substance).

[0202] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The solid was then filtered to obtain sodium thiooxapzin (white solid, 470 mg, y=47%).

[0203] 1 H NMR (300 MHz, Methanol-d4) δ 7.45 (dddd, J = 56.6, 7.8, 4.8, 2.1Hz, 10H), 3.18 (s, 4H).

[0204] 13 CNMR(75MHz.Methanol-d4) δ218. 16, 163.98, 145.30, 134.48, 132.18,128.67, 128.39, 128.31, 128.01,127.83,126.12,46.65,25.42.

[0205] Example 26 Synthesis of compound ZYSA12:

[0206]

[0207] 3-Indolepropionic acid (3 mmol, 567 mg, 1 eq) and Lawson's reagent (1.8 mmol, 728 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (15 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 100 °C. The reaction was monitored by TLC. After about 30 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0208] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The solid was then filtered to obtain sodium thio-3-indolepropionate (white solid, 438 mg, y=64%).

[0209] 1 H NMR (300 MHz, Methanol-d4) δ 7.57 (dt, J = 7.7, 1.0 Hz, 1H), 7.29 (dt, J = 8.1, 1.0 Hz, 1H), 7.08 – 6.93 (m, 3H), 3.11 – 2.96 (m, 4H).

[0210] 13 C NMR (75 MHz, Methanol-d4) δ 223.45, 138.06, 128.74, 122.55,122.03, 119.37, 119.23, 116.05, 111.99, 53.02, 24.54.

[0211] Example 27 Synthesis of compound ZYSA13:

[0212]

[0213] p-Chlorophenoxyisobutyric acid (4 mmol, 858 mg, 1 eq) and Lawson's reagent (2.4 mmol, 970 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (20 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 100 °C. The reaction was monitored by TLC. After about 20 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (pink oily substance).

[0214] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thiochlorophenoxyisobutyrate (white solid, 612 mg, y=60%).

[0215] 1 H NMR (300 MHz, Methanol-d4) δ 7.12 (d, J = 9.0 Hz, 2H), 6.86 (d, J= 9.0 Hz, 2H), 1.61 (s, 6H)

[0216] 13 C NMR (75 MHz, Methanol-d4) δ 223.38, 156.36, 129.37, 126.33,121.40, 88.18, 27.92.

[0217] Example 28 Synthesis of compound ZYSB04:

[0218]

[0219] Eicosanoid (2 mmol, 625 mg, 1 eq) and Lawson's reagent (1.2 mmol, 242 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (10 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 95 °C. The reaction was monitored by TLC. After about 20 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0220] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thioeicosate (white solid, 428 mg, y=61%).

[0221] 1H NMR (300 MHz, DMSO-d6) δ 2.29 (t, J = 7.6 Hz, 2H), 1.43 (s, 2H), 1.23 (s, 32H), 0.85 (s, 3H).

[0222] 13C NMR (75 MHz, DMSO-d6) δ 217.09, 51.30, 31.41, 29.18, 28.85,27.30, 22.19, 13.95.

[0223] Example 29 Synthesis of compound ZYSB05:

[0224]

[0225] Hexadecanoic acid (3 mmol, 769 mg, 1 eq) and Lawson's reagent (1.8 mmol, 728 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (15 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 95 °C. The reaction was monitored by TLC. After about 20 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (pink oily substance).

[0226] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thiohexadecanoate (white solid, 675 mg, y=76%).

[0227] 1 H NMR (300 MHz, DMSO-d6) δ 2.30 (t, J = 7.5 Hz, 2H), 1.48 – 1.39 (m,2H), 1.23 (s, 24H), 0.89 – 0.81 (m, 3H).

[0228] 13 C NMR (75 MHz, DMSO-d6) δ 216.68, 51.34, 31.35, 29.13, 28.97, 28.77,27.26, 22.15, 13.98.

[0229] Example 30: Synthesis of compound ZYSB06:

[0230]

[0231] Myristic acid (3 mmol, 685 mg, 1 eq) and Lawson's reagent (1.8 mmol, 728 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (15 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 95 °C. The reaction was monitored by TLC. After about 15 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0232] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thiotetradecanoate (white solid, 497 mg, y=62%).

[0233] 1 H NMR (300 MHz, Methanol-d4) δ 2.64 – 2.58 (m, 2H), 1.63 (t, J = 7.4Hz, 2H), 1.28 (s, 20H), 0.95 – 0.84 (m, 3H).

[0234] 13 C NMR (75 MHz, Methanol-d4) δ 224.09, 52.32, 33.09, 30.79, 30.50, 30.42, 29.00, 23.75, 14.48.

[0235] Example 31 Synthesis of compound ZYSB07:

[0236]

[0237] Dodecanoic acid (3 mmol, 600 mg, 1 eq) and Lawson's reagent (1.8 mmol, 728 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (15 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 95 °C. The reaction was monitored by TLC. After about 15 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0238] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The product was then filtered to obtain sodium thiodododecanoate (white solid, 411 mg, y=57%).

[0239] 1 H NMR (400 MHz, DMSO-d6) δ 2.33 – 2.27 (m, 2H), 1.44 (p, J = 7.3 Hz,2H), 1.23 (s,16H), 0.87 – 0.83 (m, 3H).

[0240] 13C NMR (75 MHz, DMSO-d6) δ 51.26, 31.36, 29.15, 28.95, 28.78, 27.25,22.15, 13.99.

[0241] Example 32 Synthesis of compound ZYSB08:

[0242]

[0243] Adamantane carboxylic acid (4 mmol, 720 mg, 1 eq) and Lawson's reagent (2.4 mmol, 970 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (20 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 90 °C. The reaction was monitored by TLC. After about 10 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (a pink oily substance).

[0244] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The solid was then filtered to obtain sodium thioadamantate (white solid, 525 mg, y=60%).

[0245] 1H NMR (300 MHz, Methanol-d4) δ 1.99 – 1.92 (m, 9H), 1.71 (t, J = 2.9Hz, 6H).

[0246] 13 C NMR (75 MHz, Methanol-d4) δ 229.92, 51.23, 42.43, 38.05, 30.40.

[0247] Example 33 Synthesis of compound ZYSB09:

[0248]

[0249] Cypermethrin (4 mmol, 568 mg, 1 eq) and Lawson's reagent (2.4 mmol, 970 mg, 0.6 eq) were added to a two-necked round-bottom flask (oven-dried) equipped with a magnetic stir bar. After replacing the nitrogen with nitrogen, toluene (20 mL) was added as a solvent using a syringe. The reaction was carried out with stirring at 90 °C. The reaction was monitored by TLC. After about 10 min, the system changed from turbid to clear, indicating that the reaction was approximately complete. The reaction solution was removed, and toluene was removed under vacuum. The residue was dissolved in DCM and loaded onto a silica gel column. Eluting with DCM yielded the crude product (pink oily substance).

[0250] The crude product obtained from the previous column chromatography step was placed in a round-bottom flask equipped with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The flask was then placed in an ice bath to cool it down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. The reaction solution was concentrated under vacuum, and anhydrous diethyl ether was added. A white solid precipitated out. The solid was then filtered to obtain sodium thiocyanate (white solid, 481 mg, y=66%).

[0251] 1 H NMR (300 MHz, Methanol-d4) δ 2.17 (s, 1H), 1.17 (d, J = 14.5 Hz,12H).

[0252] 13 C NMR (75 MHz, Methanol-d4) δ 218.04, 52.65, 32.03, 24.27, 17.13.

[0253] Example 34 Compound ZYSC03

[0254]

[0255] Following the synthesis method of target ZYSA01, crude thio-p-methoxynaphtholic acid was synthesized from p-methoxynaphtholic acid and Lawson's reagent, and then its sodium salt was formed to obtain sodium thio-p-methoxynaphtholic acid, a pale yellow solid.

[0256] 1 H NMR (300 MHz, Methanol-d4) δ8.61 (s, 1H), 8.19 (dd, J = 8.6, 1.8Hz, 1H), 7.82 (d, J = 9.0 Hz, 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.23 (d, J = 2.5Hz, 1H), 7.12 (dd, J = 9.0, 2.5 Hz, 1H), 3.91 (s, 3H).

[0257] 13 C NMR (75 MHz, Methanol-d4) δ214.25, 160.13, 140.92, 137.48, 131.78,129.52, 129.24, 127.30, 126.54, 119.87, 106.47, 55.78.

[0258] Results data:

[0259] Example 35 Determination of the half-life of hydrogen sulfide release

[0260] 1. High-performance liquid chromatography (HPLC) for the determination of PLE hydrolysis and formation of corresponding carboxylic acids in ZYSA series thiocarboxylic acids.

[0261] Add 4.5 mL of porcine liver esterase (PLE) (3 U / mL) in PBS buffer to a 20 mL reaction flask, then add 0.5 mL of ZYSA in PBS buffer (5 mM). Incubate at 37°C. At each time point, take an equal volume of 200 μL of the solution and transfer it to a 1.5 mL centrifuge tube containing 200 μL of chromatographic acetonitrile. Transfer the tube to a -20°C freezer for rapid freezing, centrifuge for 10 min (13.0 x 1000 r / min), and collect the supernatant for HPLC detection.

[0262] 2. Methylene blue method for determining hydrogen sulfide release from ZYSA series thiocarboxylic acids.

[0263] ZYSA was dissolved in 5 mL of PBS buffer (pH 7.4, final concentration 500 μM) and co-incubated with 3 U / mL PLE. At each time point, equal volumes of 200 μL of the solution were transferred to 1.5 mL centrifuge tubes containing 200 μL of zinc acetate (1%, w / v) and centrifuged for 10 minutes (13.0 x 1000 r / min). The supernatant was collected, and the precipitate was washed with PBS (100 μL × 2). Then, 600 μL of N,N-dimethyl-1,4-phenylenediamine sulfate (0.2% w / v dissolved in 20% H₂SO₄ solution) and 50 μL of ferric chloride (10% w / v dissolved in 0.2% H₂SO₄ solution) were added to the centrifuge tubes. After incubation at 37°C in a metal bath for 30 min, the absorbance at 740 m was measured using a UV-Vis spectrophotometer. The concentration of hydrogen sulfide was calculated based on the sodium hydrosulfide standard curve.

[0264] The results are shown in Table 1. Using the methylene blue method, the compounds described in this invention all showed detectable hydrogen sulfide generation. Under esterase conditions, they all released hydrogen sulfide at a certain rate. The specific release rate (t)... 1 / 2 As shown in Table 1.

[0265] Table 1. Structure, nomenclature, and half-life of thiocarboxylic acids releasing hydrogen sulfide in the embodiments of this patent.

[0266]

[0267]

[0268]

[0269] Example 36: Performance data of compound ZYSA01 prepared in Example 1

[0270] 1. Measurement of infarct area

[0271] Mouse brains were frozen at -20°C for 20 minutes and cut into 5 sections, each 1 mm thick. Each section was immediately stained with 1% 2,3,5-triphenyltetrazolium chloride (TTC) solution at 37°C for 30 minutes and fixed in 4% paraformaldehyde (PFA) solution. Infarcted areas were unstained, while normal areas were stained red. Figure 1 A). Then, images were taken of each pair of slices, and the infarct area was calculated using ImageJ. The formula is as follows: Infarct volume = Sum of infarct areas / Total area ( Figure 1 B). The results show that ZYSA01 is significantly superior to benzoic acid.

[0272] 2. Measurement of serum LDH levels and brain tissue LDA levels

[0273] After blood collection, serum was collected by centrifugation at 3000g for 10 minutes at 4°C. The method for LDH measurement is detailed in the kit instructions (Solarbio, Beijing). Brain tissue was collected to measure MDA levels, using the MDA kit instructions (Solarbio, Beijing). LDH levels reflect cell membrane damage caused by ischemia-reperfusion injury, while MDA levels reflect membrane lipid peroxidation caused by ischemia-reperfusion injury. The study found that ZYSB21 showed greater potential in the MCAO model (**P<0.01). Figure 1 C, Figure 1 D.

[0274] Example 37 Activity data of compound ZYSH06 (telmisartan)

[0275] 1. Verification of cerebral infarction activity

[0276] A mouse model of middle cerebral artery occlusion was established using 25-30 g C57 / 6J mice. We selected telmisartan as the control group and telmisartan as the treatment group, with a dosage of 0.5 × 10⁻⁶ g. -4 All other groups were subjected to a middle cerebral artery occlusion model. Anesthesia was induced with 3% isoflurane and maintained with 0.6% isoflurane. After disinfection, a midline 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 cerebral artery (MCA) was blocked in the middle. Administered medication by gavage during infarction. One hour and 20 minutes after infarction, the suture was removed and reperfusion was performed. Other treatments were the same as above. Blood was collected 24 hours later, mice were euthanized by cervical dislocation, and brain tissue was collected for subsequent experiments after craniotomy.

[0277] 2. Infarct area

[0278] Mouse brains were frozen at -20°C for 20 minutes and cut into 5 pieces, each 1 mm thick. Each slice was immediately stained with 1% 2,3,5-triphenyltetrazol chloride (TTC) solution at 37°C for 30 minutes and fixed in 4% paraformaldehyde (PFA) solution. Infarcted areas were unstained, while normal areas were stained red. Each pair of slices was then photographed, and the infarct area was calculated using ImageJ. The formula is as follows: Infarct volume = Sum of infarct areas / Total area. The results showed that both the positive control drug and the prodrug significantly reduced the infarct area, exhibiting a dose-dependent effect. ZYSA06 was significantly more effective than the positive control drug, with ZYSA06 showing greater superiority in reducing infarct area. Figure 2 A, 2B.

[0279] 3. Serum LDH levels and brain tissue MDA levels

[0280] After blood collection, serum was collected by centrifugation at 3000g for 10 minutes at 4°C. The method for LDH measurement is detailed in the kit instructions (SolarBio, Beijing). Brain tissue was collected to measure MDA levels, using the MDA kit instructions (SolarBio, Beijing). LDH levels reflect cell membrane damage caused by ischemia-reperfusion injury, while MDA levels reflect membrane lipid peroxidation caused by ischemia-reperfusion injury. Studies have shown that the ZYSA06 MCAO model exhibits greater potential (0.25 × 10⁻⁶). -4 ZYSA09 at mol / kg significantly reduced MDA levels, with a significant difference observed in the telmisartan group *P<0.05. Figure 2 C, 2D.

[0281] Example 38: Activity data of compound ZYSH21 (diclofenac)

[0282] 1. ZYSH21 releases the active drug DCF in rats.

[0283] Five rats were used, fasted for 10 hours before administration, and given free access to water. They were administered 10.50 mg / kg ZYSH21 solution, and blood samples were collected at 0 h and at 0.033, 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 6, and 8 h after administration. 5 μL of IS solution (1000 ng / ml naproxen) was added to 50 μL of plasma sample in centrifuge tubes. After vortexing for 10 seconds, 200 μL of methanol was added to the mixture to precipitate proteins. The samples were vortexed for 1 minute and centrifuged at 12000 rpm for 5 minutes at 4˚C. Finally, the supernatant was analyzed using a triple quadrupole liquid chromatography-mass spectrometry (LC-MS) system. Figure 3 )

[0284] 2. Anti-inflammatory effect of ZYSH21

[0285] Wistar rats were divided into two groups of eight each: ZYSH21 (10 mg / kg / day) and DCF (10 mg / kg / day). Immunograde natural type II collagen was dissolved in 0.05 mol / L acetic acid to prepare a 2 mg / ml solution. The type II collagen was homogenized at 4°C using Freund's incomplete adjuvant (1:1) to prepare an emulsion, with a final CII concentration of 1 mg / ml. The emulsion was thoroughly emulsified in an ice bath using a homogenizer until it did not diffuse when added to water. The emulsified mixture was injected subcutaneously at the base of the tail at a dose of 0.2 ml / rat (200 μg CII / rat). Seven days later, 0.1 ml / rat (100 μg CII / rat) was injected subcutaneously at multiple points on the back as a booster injection. Peak paw swelling was observed after 14 days. The treatment continued for 28 days, and changes in paw swelling were recorded. The paw swelling rate was assessed from day 0 to day 36. Compared with the normal group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Figure 4 ).

[0286] 3. Gastric safety of ZYSH21

[0287] Wistar rats were divided into two groups of eight: ZYSH21 (10 mg / kg / day, 5 mg / kg / day) and DCF (10 mg / kg / day, 5 mg / kg / day). All rats were administered the drugs for 28 consecutive days. ZYSH21 (10 mg / kg / day, 5 mg / kg / day) resulted in a very low ulceration rate, while DCF (10 mg / kg / day, 5 mg / kg / day) showed obvious ulceration points. Figure 5 ).

[0288] 4. ZYSH21 Acute Liver Injury

[0289] C57BL / 6JNifdc mice were fasted for 10 hours before administration, but had free access to water. Mice were randomly divided into three groups: a saline control group, a DCF group, and a ZYSH21 group. DCF was administered at a dose of 150 mg / kg, and ZYSH21 was administered at an equimolar dose. Six mice were in each group. Administration was via intraperitoneal injection over 24 hours. Blood was collected from the eyeballs at 24 hours, and the mice were euthanized by spinal dislocation. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured. Figure 6 ).

[0290] 5. ZYSH21-induced kidney damage

[0291] Wistar rats were divided into two groups of eight each: ZYSH21 (10 mg / kg / day) and DCF (10 mg / kg / day), administered for 28 consecutive days. The activity of uric acid (UA), a key indicator of renal impairment, was significantly reduced and remained within the normal range (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Figure 7 ).

[0292] Example 39 ZYSH31 (Naproxen) Activity Data

[0293] 1. PLE enzymatic hydrolysis of ZYSH31 releases the active drug NPX.

[0294] Take one 5 mL sample vial, add 50 μL of 10 mM ZYSH31 solution, 4950 μL of PBS buffer, and 1 mg of PLE to the vial, seal the vial with film, and incubate the vial in a 37°C water bath. At regular intervals, take 200 μL of the solution, filter it, and place it in a 1.5 mL injection vial. Analyze the components in the solution using a suitable HPLC mobile phase and elution. 1 / 2 It was 208.4 ± 2.1 min ( Figure 8 ).

[0295] 2. ZYSH31 releases the active drug NPX in rats.

[0296] Five rats were used. They were fasted for 10 hours before administration, but had free access to water. They were given 10.60 mg / kg ZYSH31 solution. Blood samples were collected at 0 h and at 0.033, 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 6, and 8 h after administration. 5 μL of IS solution (1000 ng / ml diclofenac) was added to 50 μL of plasma sample in a centrifuge tube. After vortexing for 10 seconds, 200 μL of methanol was added to the mixture to precipitate proteins. The sample was vortexed for 1 minute and then incubated at 12000 r·min at 4˚C. -1 Centrifuge for 5 minutes, and finally use a triple quadrupole liquid chromatography-mass spectrometry (LC-MS) system to analyze the supernatant. Figure 9 ).

[0297] 3. Anti-inflammatory effect of ZYSH31

[0298] Wistar rats were divided into two groups of six each: ZYSH31 (10 mg / kg / day) and NPX (10 mg / kg / day). Immunograde natural type II collagen was dissolved in 0.05 mol / L acetic acid to prepare a 2 mg / ml solution. The type II collagen was homogenized at 4°C using Freund's incomplete adjuvant (1:1) to prepare an emulsion, with a final CII concentration of 1 mg / ml. The emulsion was thoroughly emulsified in an ice bath using a homogenizer until it did not diffuse when added to water. The emulsified mixture was then injected subcutaneously at the base of the tail at a dose of 0.2 ml / rat (200 μg CII / rat). Seven days later, 0.1 ml / rat (100 μg CII / rat) was injected subcutaneously at multiple points on the back as a booster injection. Peak paw swelling was observed after 14 days. The treatment continued for 28 days, and changes in paw swelling were recorded. The paw swelling rate was assessed from day 0 to day 36. Compared with the normal group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Figure 10 ).

[0299] 4. Gastric safety of ZYSH31

[0300] Wistar rats were divided into two groups: ZYSH31 (10 mg / kg / day, 5 mg / kg / day) and NPX (10 mg / kg / day, 5 mg / kg / day), with six rats in each group. The rats were administered the drugs for 28 consecutive days. ZYSH31 (10 mg / kg / day, 5 mg / kg / day) resulted in a very low ulceration rate, while NPX (10 mg / kg / day, 5 mg / kg / day) showed obvious ulceration points. Figure 11 A).

[0301] 5. ZYSH31 Acute Liver Injury

[0302] C57BL / 6JNifdc mice were fasted for 10 hours before administration, but had free access to water. Mice were randomly divided into three groups: a saline control group, an NPX group, and a ZYSH31 group. NPX was administered at a dose of 150 mg / kg, and ZYSH31 was administered at an equimolar dose, with 6 mice in each group. Administration was via intraperitoneal injection over 24 hours. Blood was collected from the eyeballs at 24 hours, and the mice were euthanized by spinal dislocation. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured. Figure 11 B).

[0303] 6. ZYSH31 kidney damage

[0304] Wistar rats were divided into two groups of six each: ZYSH31 (10 mg / kg / day) and NPX (10 mg / kg / day), administered for 28 consecutive days. The activity of albumin, a renal function indicator, was abnormally elevated, remaining within the normal range (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Figure 12 ).

[0305] Example 40: Aquatic Environment Stability

[0306] ZYSA01-6, ZYSA12-ZYSA13, ZYSB01-3, ZYSC01-02, ZYSH01-ZYSH05, ZYSH31, and ZYSH21 were dissolved in PBS solution to a final concentration of 500 μM. Their stability was observed at room temperature. The stability of ZYSA01-6, ZYSA12-ZYSA13, ZYSB02-3, ZYSC01-02, ZYSH01-ZYSH05, ZYSH31, and ZYSH21 was monitored by HPLC, and the stability of ZYSB01 was monitored by UV. The study found that ZYSA01-6, ZYSA12-ZYSA13, ZYSB01-3, ZYSC01-02, ZYSH01-ZYSH05, ZYSH31, and ZYSH21 all exhibited good stability, with no significant decomposition observed after 144 hours.

Claims

1. A hydrogen sulfide prodrug, characterized in that, It has the structure shown in equation (III): Among them, in compound (III): Z + Choice Na + , K + H + ; L2 is selected from covalent bonds, -(CH2). n - or C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4; R7 is selected from H and C1-C6 alkyl groups; R8 is selected from H, -C(=O)-R 10 C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; R 10 Selected from substituted or unsubstituted C6-C 10 The aryl or substituted or unsubstituted C3-C10 heteroaryl group, wherein the substituent is selected from H, halogen, -OH, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, and C1-C4 haloalkyl; R9 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy.

2. The hydrogen sulfide prodrug according to claim 1, characterized in that, In compounds of formula (III): L2 is selected from -(CH2). n -; n is selected from 0, 1, 2 or 3.

3. The hydrogen sulfide prodrug according to claim 1, characterized in that, In the compound of formula (III): R7 is selected from H, C1-C4 alkyl; preferably, R7 is selected from methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

4. The hydrogen sulfide prodrug according to claim 1, characterized in that, In compounds of formula (III): R8 is selected from H or -C(=O)-R 10 ;R 10 The following groups, whether substituted or unsubstituted, are selected: phenyl, naphthyl, anthraquinyl, oxazolyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolinyl; the substituents are selected from H, halogen, -OH, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

5. The hydrogen sulfide prodrug according to claim 1, characterized in that, In compounds of formula (III): R9 is selected from H, halogen, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy or C1-C4 haloalkyl.

6. A hydrogen sulfide prodrug having the following structure: 。 7. A pharmaceutical composition comprising a hydrogen sulfide prodrug as described in any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

8. Use of the hydrogen sulfide prodrug according to any one of claims 1 to 6 in the preparation of a medicament for treating cardiovascular and cerebrovascular diseases, anti-inflammatory, antipyretic and analgesic, rheumatoid arthritis, gout, lowering blood pressure, lowering blood lipids, or reducing hepatotoxicity caused by drug-induced GSH consumption or mitochondrial damage.

9. A method for forming H2S, wherein the hydrogen sulfide prodrug according to any one of claims 1 to 6 is contacted with an enzyme, and the thiocarboxylic acid structure is converted into a carboxylic acid structure under enzyme catalysis and H2S is released.

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