Prodrug design strategy for releasing hydrogen sulfide based on direct esterase hydrolysis of thiocarboxylic acid and application of prodrug design strategy in carboxylic acid drug modification

Abstract

The invention discloses a design strategy of a prodrug for releasing hydrogen sulfide based on direct esterase hydrolysis of thiocarboxylic acid and application of the prodrug in carboxylic acid drug modification, according to the hydrogen sulfide prodrug, an existing drug with a carboxylic acid structure is vulcanized to form the thiocarboxylic acid structure, and the thiocarboxylic acid structure can release H2S under the 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

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a prodrug design strategy based on direct esterase hydrolysis of thiocarboxylic acid to release hydrogen sulfide and its application in the modification of carboxylic acid drugs. Background Art

[0002] Hydrogen sulfide (H2S) was considered a toxic gas with a rotten egg smell in the past, but recent studies have found that H2S is the third gas signal molecule besides CO and NO, and is involved in a variety of physiological and pathological processes. Studies have shown that H2S can dilate blood vessels and lower blood pressure, resist oxidative stress and reduce tissue damage, regulate intestinal flora, and protect mucosa. However, high concentrations of H2S are toxic. In order to exert the characteristics of H2S as a gas signal molecule, it must be able to be released slowly and in a controlled manner, so it is necessary to construct H2S prodrugs.

[0003] At present, a variety of hydrogen sulfide prodrugs have been developed, mainly including natural hydrolysis type: inorganic sulfide salts (Na2S, NaHS, CaS, etc.), Lawsson reagent derivatives (GYY4137), response trigger type, etc.:

[0004] The release mechanism of the esterase-responsive trimethyl-locked cyclization prodrug that releases hydrogen sulfide is that the esterase hydrolyzes the ester bond in the structure, and the generated intermediate releases hydrogen sulfide and lactone through a cyclization reaction. However, the esterase-responsive hydrogen sulfide prodrug has an overly complex structure and its pharmacokinetic properties in vivo are unknown, which is not conducive to clinical application.

[0005] The release mechanism of COS-based hydrogen sulfide prodrug is to release COS in response to ROS, esterase, pH, thiol, light and other signals in the body. COS is further hydrolyzed to release hydrogen sulfide and CO2 under the action of carbonic anhydrase. COS itself is cytotoxic and has low prospects for biological applications.

[0006] The release mechanism of hydrogen sulfide prodrugs based on thioester bonds is that thiols in the body attack the thioester bonds, thereby releasing sulfide, which further undergoes sulfur exchange reaction with thiols to further release hydrogen sulfide. Thiol-responsive hydrogen sulfide prodrugs have different metabolic pathways in vivo and in vitro, and under conditions of high oxidative stress in vivo, thiols are often oxidized to form disulfide forms, which is not conducive to drug release.

[0007] The pH-sensitive H2S prodrug of thiophosphates, under acidic conditions, the nucleophilic carboxylic acid attacks the phosphate center to form a stable five-membered ring, accompanied by the release of H2S. The structure of the compound is complex, which is not conducive to its application in drugs, and the drug metabolism pathway in the body is unknown.

[0008] The release mechanism of geminal dithiol hydrogen sulfide prodrug is to protect unstable geminal dithiol with a photosensitive group (2-nitrobenzyl) or a carbonyl group. Under ultraviolet light or when thiol attacks, geminal dithiol is released, and then hydrolysis reaction occurs to release H2S. It needs to be stimulated by thiol in the body to release hydrogen sulfide, which means that its effect may be limited by thiol-rich tissues and organs. Under high oxidative stress, thiol may be oxidized to form disulfide, which is not conducive to drug release.

[0009] The release mechanism of thioamino acid hydrogen sulfide prodrugs is to use the thioamino acid to react with bicarbonate to form a carbamate intermediate, which then undergoes a cyclization reaction to generate N-carboxylic anhydride accompanied by the release of H2S. The drug metabolism in the body may be inconsistent with the assumption.

[0010] Arylsulfamide prodrugs, whose reported mechanism is natural slow hydrolysis or high concentration of thiol attack accelerated hydrolysis to release hydrogen sulfide. Lack of organ or tissue specific release mode, uncertainty of efficacy and dosage, and possible adverse reactions at high doses, and the compound structure is too complex to be integrated into drugs.

[0011] It also includes natural polysulfides and ADT-OH whose release mechanism is unclear.

[0012] Due to the problems of the aforementioned hydrogen sulfide prodrugs, no hydrogen sulfide prodrug has entered clinical use. In addition, many drugs used in clinical practice have poor efficacy or serious toxic side effects. For example, the natural hydrolysis type will undergo hydrolysis to generate hydrogen sulfide in an aqueous environment, and its release is uncontrollable and cannot meet the fixed-point and quantitative release, so its application is limited. The chemical structure of other responsive release types is too complex, and it is difficult to accurately determine whether it is the prodrug carrier or H2S itself that exerts its efficacy when using the drug, and its pharmacokinetic and pharmacological properties are unknown; and for thiol-sensitive H2S prodrugs, research results show that when the human body is ischemic reperfusion injury or inflammation, the internal environment is in a highly oxidized state, and free thiols are often in the form of disulfide, making it difficult to truly achieve thiol-based prodrug activation. The cardiovascular drug butylphthalide often needs to be used in combination with other drugs in clinical use because of its slow effect; long-term use of non-steroidal anti-inflammatory drugs such as diclofenac sodium and aspirin will lead to serious gastrointestinal, liver, and kidney toxic side effects.

[0013] In summary, the hydrogen sulfide prodrugs reported in current studies have poor drugability and are difficult to use clinically. Summary of the invention

[0014] In view of the problems of existing drugs, the present invention aims to prepare more stable and effective hydrogen sulfide prodrugs.

[0015] In the first aspect, the hydrogen sulfide prodrug provided by the present invention is obtained by sulfiding an existing drug having a carboxylic acid structure to form a thiocarboxylic acid structure, which can release H2S under enzyme catalysis conditions.

[0016] The second invention is that 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 forms a carboxylic acid structure under enzyme catalysis conditions and releases H2S.

[0017] The enzymes described in the present invention may include any enzyme that can decompose the thiocarboxylic acid structure into a carboxylic acid structure and release H2S. The inventors have found through various in vivo and in vitro experiments that various enzymes in vivo can achieve this function. For example, the inventors injected the prodrug having a thiocarboxylic acid structure of the present invention through the tail vein. The pharmacokinetic study results show that the generation of carboxylic acid can be observed when the drug is injected into the tail vein. In the embodiments of this patent, only pig liver esterase (PLE) is used as an example, but it is not limited to this enzyme.

[0018] The present invention proves for the first time that a compound with a thiocarboxylic acid structure can be hydrolyzed by enzymes under enzyme catalysis to release H2S, and that by using thiocarboxylic acids as a new type of carrier for hydrogen sulfide, the hydrolysis rate of thiocarboxylic acids can be controlled by esterase, thereby slowly releasing hydrogen sulfide. Compared with some existing naturally hydrolyzed precursors that can be hydrolyzed to generate hydrogen sulfide in an aqueous environment, the compound of the present invention can remain stable in an aqueous environment without the addition of enzymes, and can only slowly release H2S by enzyme catalysis, thereby achieving a slow and controllable release of H2S in treatment, avoiding the uncontrollable risk caused by the rapid local release of hydrogen sulfide donors during use. This discovery is of great significance in the field of biomedicine.

[0019] Specifically, the thiocarboxylic acid structure in the prodrug of the present invention releases hydrogen sulfide in the following manner:

[0020]

[0021] In a third aspect, the present invention provides a series of hydrogen sulfide prodrugs to demonstrate the aforementioned aspects of the present invention.

[0022] The present invention uses thiocarboxylic acid for the preparation of hydrogen sulfide prodrugs for the first time. The inventors have found that drugs with carboxylic acid structures in active drugs can be sulfided with carboxylic acid structures to form drugs with thiocarboxylic acid structures of the present invention. In certain embodiments, the hydrogen sulfide prodrug provided by the present 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 and analgesic drugs, rheumatic or rheumatoid arthritis drugs, anti-gout drugs, blood pressure lowering, blood lipid lowering or drugs for reducing GSH consumption or liver toxicity caused by mitochondrial damage, etc.; in some examples, for example, the following types of drugs with -COOH structure: clofibric acid lipid regulating drugs, sartan drugs, sulfonamide drugs, indole drugs, non-steroidal anti-inflammatory drugs, etc. In some more specific examples, these drugs include but are not limited to diclofenac, ketoprofen, indolepropionic acid, indomethacin, telmisartan, gemfibrozil, naproxen, oxaprozin, ibuprofen, flurbiprofen, probenecid, etc. Other drugs with carboxylic acid structures can also be sulfided to form the precursor compounds of the present invention, and the effect of releasing hydrogen sulfide can be achieved.

[0023] In some examples, the hydrogen sulfide prodrug described in the present invention is obtained by sulfiding an existing sartan drug having a -COOH structure to form a thiocarboxylic acid structure, which can release H2S under enzyme catalysis conditions; the sartan drug is any sartan drug having a -COOH structure used clinically, such as valsartan, eprosartan, and telmisartan; when containing multiple -COOH structures, any one or more -COOH can be sulfided to form a thiocarboxylic acid structure.

[0024] In other embodiments, the hydrogen sulfide prodrug of the present invention has a structure as shown in the following formula (I), formula (II), formula (III) or (IV):

[0025]

[0026] Among them, in the compound of formula (I):

[0027] Z + Selected from Na + , K + , H + ;

[0028] A is selected from C6-C 20 Aryl or C3-C 20 Heteroaryl; preferably C6-C 10 Aryl or C3-C 10Heteroaryl; more preferably, the following groups: phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, purinyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, indolyl, benzotriazolyl, benzothienyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl or carbazolyl;

[0029] L1 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl, -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-, -O-(CH2) n -、-O-(CH2) n -O-;

[0030] n is selected from 0 or an integer from 1 to 8;

[0031] R1 and R2 are independently selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -SO2NR3R4-, C6-C 10 Aryl or C3-C 10 heteroaryl;

[0032] R3, R4, R5, R6 are independently selected from H or C1-C6 alkyl;

[0033] Among them, in the compound of formula (II):

[0034] Z + Selected from Na + , K + , H + ;

[0035] R' is selected from the following substituted or unsubstituted groups: C1-C 30 Alkyl, C3-C 10 Cycloalkyl, C8-C 16 Bridged cycloalkane, C6-C 16 Spirocycloalkane; the substituent is one or more, and the substituent is selected from H, halogen, -OH, CN, NO2, C1-C6 alkyl;

[0036] Among them, in the compound of formula (III):

[0037] Z + Selected from Na + , K + , H + ;

[0038] L2 is selected from a covalent bond, -(CH2)n - or C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4;

[0039] R7 is selected from H, C1-C6 alkyl;

[0040] 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 aryl or substituted or unsubstituted C3-C 10 Heteroaryl, substituents selected from H, halogen, -OH, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl;

[0041] R9 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy;

[0042] Among them, in the compound of formula (IV):

[0043] Z + Selected from Na + , K + , H + ;

[0044] L3 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4;

[0045] X is selected from -NH- or -C(=O)-;

[0046] R a , R b Independently selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy.

[0047] Furthermore, in the compound of formula (I):

[0048] In some specific embodiments, A is selected from C6-C 10 Aryl or C3-C 10 Heteroaryl; A is further preferably the following groups: phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, purinyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, indolyl, benzotriazolyl, benzothienyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl or carbazolyl, etc.

[0049] In some specific embodiments, L1 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl, -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-; in some more specific embodiments, L1 is selected from a covalent bond, -(CH2) n -、-CH(CH3)-、 -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-.

[0050] In some specific embodiments, n is selected from 0, 1, 2, 3 or 4.

[0051] In some specific embodiments, R1 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C6-C 10 Aryl or C3-C 10 Heteroaryl; R1 is further preferably the following groups: H, halogen, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl.

[0052] In some specific embodiments, R2 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -SO2NR3R4-, C6-C 10 Aryl or C3-C 10 Heteroaryl; R2 is further preferably the following groups: H, halogen, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, -CH2Cl, -CF3, -CH2CF3, -CH2CCl3, -SO2NR3R4-, phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl.

[0053] In some specific embodiments, R3, R4, R5, and R6 are independently selected from C1-C6 alkyl; further, R3, R4, R5, and R6 are independently selected from methyl, ethyl, propyl, isopropyl, butyl, and tert-butyl.

[0054] Furthermore, in the compound of formula (II):

[0055] In some specific embodiments, R' is selected from the following substituted or unsubstituted groups: C2-C6 branched alkyl, C1-C 20 Straight chain alkyl, C3-C6 cycloalkyl, C8-C 16The substituents are one or more, and the substituents are selected from H, halogen, -OH, CN, NO2, and C1-C4 alkyl.

[0056] Furthermore, in the compound of formula (III):

[0057] In some specific embodiments, L2 is selected from -(CH2) n -; n is selected from 0, 1, 2 or 3.

[0058] In some specific embodiments, R7 is selected from H, C1-C4 alkyl; in some specific examples, R7 is selected from methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

[0059] In some specific embodiments, R8 is selected from H or -C(=O)-R 10 ; R 10 Selected from the following substituted or unsubstituted groups: phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl; the substituents are selected from H, halogen, -OH, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

[0060] In some specific embodiments, R9 is selected from H, halogen, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy or C1-C4 haloalkyl.

[0061] Further, in the compound of formula (IV):

[0062] In some specific embodiments, L3 is selected from a covalent bond, -(CH2) n -、-CH(CH3)-、 n is selected from 0, 1, 2 or 3;

[0063] In some specific schemes, R a , R b Independently selected from H, halogen, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

[0064] In some embodiments, the present invention provides the following specific hydrogen sulfide prodrugs:

[0065]

[0066]

[0067]

[0068] The hydrogen sulfide prodrug of the present invention can be prepared by sulfiding the carboxylic acid of an existing drug using chemical synthesis methods commonly used in the prior art.

[0069] The present invention also provides a pharmaceutical composition, which comprises the hydrogen sulfide prodrug of the present invention and a pharmaceutically acceptable carrier.

[0070] The hydrogen sulfide prodrug of the present invention can be administered to a subject under an environment sufficient to form hydrogen sulfide, by administering the compound of the present invention or a pharmaceutical composition containing the compound of the present invention. The inventors have experimentally found that the release of hydrogen sulfide can be achieved by oral administration, intravenous administration, and the like.

[0071] The present invention also provides the use of the hydrogen sulfide prodrug in preparing drugs for treating cardiovascular and cerebrovascular diseases, anti-inflammatory, antipyretic and analgesic, rheumatic or rheumatoid arthritis, anti-gout, lowering blood pressure, lowering blood lipids, or reducing drug consumption of GSH or liver toxicity caused by mitochondrial damage.

[0072] The compounds of the present invention may be used alone as a monotherapy or in combination with other active drugs.

[0073] The hydrogen sulfide prodrug of the present invention has the following advantages over the prior art:

[0074] (1) The different physiological functions of hydrogen sulfide can be achieved by simply changing the structure and chemical properties of carboxylic acids. In this modification process, there is no need to change the original drug skeleton, only slight changes to the drug structure, which can greatly shorten the time and cost of drug development.

[0075] (2) The hydrogen sulfide prodrug of the present invention, the thiocarboxylic acid can be hydrolyzed by esterase to release hydrogen sulfide, and the anti-inflammatory, myocardial protection, gastrointestinal protection, etc. of hydrogen sulfide are used to improve the toxic and side effects of existing drugs, while the physiological functions of hydrogen sulfide itself are exerted to achieve the clinical application of hydrogen sulfide. Based on this characteristic, the hydrogen sulfide prodrug of the present invention has the characteristics of improving the efficacy while reducing the toxic and side effects of the drug. For example, in the treatment of cerebral infarction and inflammation, the use of the thiocarboxylic acid compounds synthesized by the present invention can enhance the efficacy of the drug and reduce the adverse effects of the drug on human health.

[0076] Unless otherwise specified, the terms used in the present invention have the following definitions:

[0077] "Alkyl" means a saturated aliphatic hydrocarbon group of 1-30 carbon atoms, including straight chain and branched groups (the numerical range mentioned in this application, such as "1-20", means that the group, in this case an alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). For example, 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.

[0078] "Haloalkyl" refers to an alkyl group substituted with one or more halogen atoms which may be the same or different, for example, -CH2Cl, -CF3, -CH2CF3, -CH2CCl3, and the like.

[0079] "Alkoxy" means -O-(unsubstituted alkyl) or -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0080] "Cycloalkyl" means an all-carbon monocyclic or fused ring ("fused" ring means that each ring in the system shares an adjacent pair of carbon atoms with every other ring in the system) group in which one or more rings do not have a completely connected pi electron system. Examples of cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cycloheptatriene.

[0081] "Aryl" means an all-carbon monocyclic or fused polycyclic group of 1 to 12 carbon atoms, having a completely conjugated π electron system. Non-limiting examples of aryl are phenyl, naphthyl and anthracenyl. Aryl 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.

[0082] "Heteroaryl" means a stable monocyclic ring with up to 3-10 atoms in the ring or a bicyclic carbocyclic ring with up to 3-10 atoms in each ring, wherein at least one ring is aromatic and contains 1-4 heteroatoms selected from O, N and S. Non-limiting examples of unsubstituted heteroaryl groups include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, purinyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, indolyl, benzotriazolyl, benzothienyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl and carbazolyl. Heteroaryl can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and further preferably one or two.

[0083] "Spirocyclic": When there are two rings in a molecule and these two rings share a carbon atom, this system is called a spirocyclic.

[0084] The "bridged ring" refers to a polycyclic hydrocarbon that shares two or more carbon atoms (bridgehead carbons), and is classified into bicyclic hydrocarbons, tricyclic hydrocarbons, tetracyclic hydrocarbons, etc. according to the number of constituent rings.

[0085] "Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Figure 1The pharmacological evaluation of ZYSA01 in cerebral infarction; A is the infarct area; B is the quantitative analysis of the infarct area; C is the serum LDH level analysis; D is the tissue MDA level analysis.

[0087] Figure 2 A is the activity data of ZYSH06; A is the infarct area; B is the quantitative analysis of the infarct area; C is the serum LDH level analysis; D is the tissue MDA level analysis.

[0088] Figure 3 This is the blood concentration-time curve of the mutual conversion between ZYSH21 and DCF after oral administration of ZYSH21 (10 mg / kg).

[0089] Figure 4 This is a curve chart of the changes in toe swelling rates of rats in the DCF and ZYSH21 groups after intragastric administration of 10 mg / kg, n=8.

[0090] Figure 5 This is the percentage of gastric ulcers in rats in the DCF and ZYSH21 groups after intravenous administration of 10 mg / kg, n=8.

[0091] Figure 6 The levels of AST and ALT in serum 24 hours after administration of DCF (150 mg / kg, ip) and ZYSH21 (157 mg / kg, ip).

[0092] Figure 7 The uric acid levels in the serum of rats in the DCF and ZYSH21 groups after intragastric administration of 10 mg / kg, n=8.

[0093] Figure 8 This is the blood concentration-time curve of the mutual conversion between ZYSH31 and NPX after oral administration of ZYSH31 (10 mg / kg).

[0094] Fig. 9 This is a curve chart of the changes in toe swelling rate of rats in the NPX and ZYSH31 groups after intravenous administration of 10 mg / kg, n=6.

[0095] Fig.10 This is the percentage of gastric ulcers in rats in the NPX and ZYSH31 groups after intravenous administration of 10 mg / kg, n=6.

[0096] Fig.11 The levels of AST and ALT in serum 24 hours after administration of NPX (150 mg / kg, ip) and ZYSH31 (159 mg / kg, ip), n=6.

[0097] Fig.12The levels of serum albumin in rats of NPX and ZYSH31 groups after intravenous administration of 10 mg / kg, n=6. DETAILED DESCRIPTION

[0098] The following examples are provided for better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified.

[0099] Example 1 Compound ZYSA01

[0100]

[0101] Thiobenzoic acid (purchased from a reagent supplier) (1.00 g, 7.24 mmol, 0.85 mL) was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of methanol was added to dissolve it. The mixture was placed in an ice-salt bath to cool down. After cooling down, a methanol solution of NaOH (0.23 g, 5.80 mmol, 0.8 eq) was added, and the mixture was stirred in an ice-salt bath for 5 minutes. After the reaction solution was spin-dried, anhydrous ether was added, and a white solid was precipitated. Sodium thiobenzoate (1.15 g, 99%) was obtained by filtration.

[0102] 1 H NMR (300MHz, CD3OD) δ8.15–7.70(m,2H),7.47–6.94(m,3H).

[0103] 13 C NMR (75MHz, CD3OD) δ213.44,144.40,130.00,128.90,127.90,127.40,126.96.

[0104] Example 2 Compound ZYSA02

[0105]

[0106] Referring to the synthesis method of target ZYSA01, phenylacetic acid and Lawesson's reagent were used as raw materials to synthesize crude thiophenylacetic acid, which was then converted into sodium salt to obtain sodium thiophenylacetate as a white solid.

[0107] 1 H NMR(300MHz,DMSO-d6)δ7.08-7.19(m,5H),3.68(s,2H).

[0108] 13 C NMR (101MHz, CD3OD) δ137.89,128.85,127.64,125.64,57.34.

[0109] Example 3 Compound ZYSA03

[0110]

[0111] Referring to the synthesis method of target ZYSA01, phenylpropionic acid and Lawesson's reagent were used as raw materials to synthesize crude thiophenylpropionic acid, which was then converted into sodium salt to obtain sodium thiophenylpropionic acid, a white solid.

[0112] 1 H NMR (500MHz, DMSO-d6) δ7.22(t,J=10.0Hz,2H),7.10-7.17(m,3H),2.79(t,J=5.0Hz,2H),2.63(t,J=10.0Hz,2H).

[0113] 13 C NMR (75MHz, DMSO-d6) δ215.98,143.31,128.69,128.55,125.76,53.24,33.75.

[0114] Example 4 Compound ZYSA04

[0115]

[0116] 4-Methylbenzoic acid (1.00 g, 7.34 mmol) and Lawesson's reagent (1.63 g, 4.03 mmol, 0.55 eq) were added to a two-necked round-bottom flask with a magnetic stirrer. After replacing N2, toluene (30 mL) was added as solvent using a syringe. The mixture was stirred at 130°C for 30 min. The system changed from turbid to clear. The reaction was basically complete as monitored by TLC. The mixture was concentrated under reduced pressure to remove toluene. 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. The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of methanol was added to dissolve it. It was placed in an ice-salt bath to cool down. After cooling, a methanol solution of NaOH (0.19 g, 4.70 mmol, 0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was spin-dried, anhydrous ether was added, and a white solid was precipitated. Filter to obtain sodium 4-methylthiobenzoate (770 mg, 60%).

[0117] 1 H NMR (500MHz, DMSO-d6) δ7.98 (d, J = 10.0 Hz, 2H), 7.05 (d, J = 5.0 Hz, 2H), 2.30 (s, 3H).

[0118] 13C NMR (126MHz, DMSO-d6) δ207.96,143.21,138.62,128.47,127.74,21.32.

[0119] Example 5 Compound ZYSA05

[0120]

[0121] Referring to the synthesis method of ZYSA04, 4-methoxybenzoic acid and Lawesson's reagent were used as raw materials to synthesize crude 4-methoxythiobenzoic acid, which was then converted into sodium salt to obtain sodium 4-methoxythiobenzoate as a white solid.

[0122] 1 H NMR (500MHz, DMSO-d6) δ8.05 (d, J = 10.0 Hz, 2H), 6.77 (d, J = 10.0 Hz, 2H), 3.75 (s, 3H).

[0123] 13 C NMR (126MHz, DMSO-d6) δ207.50,160.76,138.57,130.16,112.25,55.57.

[0124] Example 6 Compound ZYSA06

[0125]

[0126] Add p-methylphenylacetic acid and Lawesson's reagent (0.55eq) to a two-necked round-bottom flask (oven dried) with a magnetic stirrer, set up a reflux tube, replace N2, heat to 110°C and start reflux, monitor the reaction by TLC, after about 1.5h of reflux reaction, the system changes from turbid to clear, and the reaction is almost complete. Take out the reaction solution, remove toluene in vacuo, dissolve the residue in DCM, load it onto a silica gel column, and elute with DCM to obtain a crude product (yellow oil).

[0127] The crude product obtained in the previous step is placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of MeOH is added to dissolve it. It is then placed in an ice-salt bath to cool. After cooling, a MeOH solution of NaOH (0.8 eq) is added, and the mixture is stirred in an ice-salt bath for 5 minutes. After MeOH is removed in vacuo, a small amount of DCM is added to the residue to dissolve it, and anhydrous ether is added to precipitate a white solid. The sodium salt of thio-p-methylphenylacetic acid can be obtained by filtration.

[0128] 1H NMR (300MHz, DMSO-d6) δ7.07 (d, J = 8.0 Hz, 2H), 6.99 (d, J = 7.8 Hz, 2H), 3.64 (s, 2H), 2.23 (s, 3H).

[0129] 13 C NMR (75MHz, DMSO) δ212.98,136.06,133.90,128.99,128.09,58.04,20.70.

[0130] Example 7 Compound ZYSA07

[0131]

[0132] Referring to the synthesis method of target ZYSA01, 4-chlorophenylpropionic acid and Lawesson's reagent were used as raw materials to synthesize crude 4-chlorothiophenylpropionic acid, which was then converted into sodium salt to obtain sodium 4-chlorothiophenylpropionic acid, a white solid.

[0133] 1 H NMR (300MHz, CD3OD) δ7.17-7.23(m,4H),2.83-2.95(m,4H).

[0134] 13 C NMR (101MHz, CD3OD) δ140.75,131.00,129.63,127.84,52.23,32.80.

[0135] Example 8 Compound ZYSA08

[0136]

[0137] Referring to the synthesis method of target ZYSA01, 4-methoxyphenylpropionic acid and Lawesson's reagent were used as raw materials to synthesize crude 4-methoxythiophenylpropionic acid, which was then converted into sodium salt to obtain sodium 4-methoxythiophenylpropionic acid as a white solid.

[0138] 1 H NMR (300MHz, CD3OD) δ7.07 (d, J = 9.0Hz, 2H), 6.77-6.80 (m, 2H), 3.70 (s, 3H), 2.69-2.74 (m, 2H), 2.55-2.59 (m, 2H).

[0139] 13 C NMR (101MHz, CD3OD) δ157.84,133.94,128.83,128.77,113.28,54.24,52.99,32.76.

[0140] Example 9 Compound ZYSB01

[0141]

[0142] γ-Butyrolactone (2 g, 23.2 mmol) and Lawesson's reagent (5.6 g, 13.9 mmol, 0.6 eq) were added to a two-necked round-bottom flask with a magnetic stirrer. After replacing N2, toluene (30 mL) was added as solvent using a syringe. The mixture was stirred at 130°C for 5 h. The system turned from turbid to clear. The reaction was basically complete as monitored by TLC. The mixture was concentrated under reduced pressure to remove toluene. Thio-γ-butyrolactone was obtained by elution using silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 5 / 1) as a yellow oily liquid (647 mg, 30%).

[0143] Thio-γ-butyrolactone (193 mg, 1.9 mmol) was added to a single-mouth round-bottom flask with a magnetic stirrer, and 10 mL of methanol was added to dissolve it. Sodium hydroxide (91 mg, 2.3 mmol, 1.2 eq) was weighed and dissolved in 3 mL of water, and added to the methanol solution of thio-γ-butyrolactone. Stir at room temperature for 3 h. The reaction was complete after TLC monitoring. The solvent was removed by concentration under reduced pressure to obtain sodium γ-hydroxythiobutyrate as a white solid (183 mg, 70%).

[0144] 1 H NMR (300MHz, CD3OD) δ3.57 (t, J = 6.0 Hz, 2H), 2.71 (t, J = 6.0 Hz, 2H), 1.82-1.91 (m, 2H). 13 C NMR (75MHz, CD3OD) δ221.88, 61.32, 34.48, 30.28.

[0145] Example 10 Compound ZYSB02

[0146]

[0147] Referring to the synthesis method of ZYSB01, α-methyl-γ-butyrolactone and Lawesson's reagent were used as raw materials to synthesize thio-α-methyl-γ-butyrolactone, which was then alkaline hydrolyzed into a sodium salt to obtain sodium α-methyl-γ-hydroxythiobutyrate as a white solid.

[0148] 1 H NMR (300MHz, CD3OD) δ3.54-3.60(m,2H),2.85-2.96(m,1H),1.85-1.98(m,1H),1.49-1.58(m,1H),1.11(d,J=6.0Hz,3H).

[0149] 13C NMR(75MHz,,CD3OD)δ226.19,60.17,50.79,37.99,18.63.

[0150] Example 11 Compound ZYSB03

[0151]

[0152] Octadecanoic acid and Lawesson's reagent (0.55 eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, dry THF was added as solvent with a syringe. The reaction was stirred at 65°C and monitored by TLC. After about one night, the system changed from turbid to clear. The reaction solution was taken out, THF was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (white solid) was obtained by eluting with DCM.

[0153] The crude product obtained in the previous step is placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM is added to dissolve it. Then, it is placed in an ice-salt bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) is added, and stirred in an ice-salt bath for 5 minutes. After removing MeOH in a vacuum, a small amount of DCM is added to the residue to dissolve it, and anhydrous ether is added. A white solid will precipitate, and sodium thiooctadecanoate can be obtained by filtration.

[0154] 1 H NMR (300MHz, DMSO-d6) δ2.30(t,J=7.5Hz,2H),1.43(t,J=7.2Hz,2H),1.23(s,28H),0.91–0.79(m,3H).

[0155] Example 12 Compound ZYSC01

[0156]

[0157] Referring to the synthesis method of target ZYSA01, naphthoic acid and Lawesson's reagent were used as raw materials to synthesize crude thionaphthoic acid, which was then converted into sodium salt to obtain sodium thionaphthoate, a light yellow solid.

[0158] 1 H NMR (300MHz, DMSO-d6) δ8.56(s,1H),8.26(d,J=9.0Hz,1H),7.84-7.96(m,2H),7.74(d,J=9.0Hz,1H),7.46-7.48(m,2H).

[0159] 13C NMR (101MHz, CD3OD) δ213.07,141.61,134.56,132.63,128.91,128.00,127.11,126.56,126.27,125.67,125.37.

[0160] Example 13 Compound ZYSC02

[0161]

[0162] Referring to the synthesis method of target ZYSA01, naphthylacetic acid and Lawesson's reagent were used as raw materials to synthesize crude thionaphthylacetic acid, which was then converted into sodium salt to obtain sodium thionaphthylacetic acid, a light yellow solid.

[0163] 1 H NMR (300MHz, DMSO-d6) δ7.78-7.83(m,2H),7.73(d,J=9.0Hz,1H),7.62(s,1H),7.36-7.46(m,3H),3.86(s,2H).

[0164] 13 C NMR (101MHz, CD3OD) δ135.52,133.67,132.26,127.60,127.20,127.11,127.08,127.02,125.30,124.76,57.46.

[0165] Example 14 Compound ZYSD01

[0166]

[0167] Phenylglycine (200 mg, 1.04 mmol, 1 eq), NHS (238 mg, 2.07 mmol, 2 eq), EDCI HCl (298 mg, 1.55 mmol, 1.5 eq) were added to a two-necked round-bottom flask with a magnetic stirrer. After replacing N2, anhydrous THF (5 mL) was added by syringe under ice bath, and the mixture was returned to room temperature. The reaction was monitored by TLC. After 12 hours of reaction, the reaction was almost complete. The reaction solution was taken out and concentrated in vacuo. The residue was dissolved with ethyl acetate (30 mL). The organic phase was washed with water (3×10 mL), and then dried with anhydrous Na2SO4. The organic phase was concentrated in vacuo to remove the solvent to obtain the active ester product (white solid, 265 mg, y=88%).

[0168] The active ester product (200 mg, 0.68 mmol, 1 eq) obtained in the previous step was dissolved in THF (5 mL) and placed in an ice bath to cool. 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 completed, 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 taken out, evaporated in vacuo, diluted with water, and the pH was adjusted to 3 with 1 M 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 in vacuo. A small amount of DCM dissolved residue was loaded onto a silica gel column and eluted with DCM containing 1% volume acetic acid. The eluate was collected, most of the solvent was removed under reduced pressure, and a large amount of PE was added to precipitate a white powdery solid. After filtration, the precipitate was washed with PE to obtain the thio product (white powder solid, 38 mg, y=20%).

[0169] 1 H NMR (300MHz, Methanol-d4) δ7.36–7.21(m,5H),4.10(s,2H),3.59(s,2H).

[0170] 13 C NMR (75MHz, MeOD) δ175.56,168.55,140.18,138.77,128.96,126.98,45.64,44.73,40.58,30.20,21.41,17.61.

[0171] Example 15 Compound ZYSD02

[0172]

[0173] Ibuprofenyl glycine (400 mg, 1.52 mmol, 1 eq), NHS (350 mg, 3.04 mmol, 2 eq), EDCI HCl (435 mg, 2.28 mmol, 1.5 eq) were added to a two-necked round-bottom flask with a magnetic stirrer. After replacing N2, anhydrous THF (8 mL) was added with a syringe in an ice bath, and the mixture was returned to room temperature. The reaction was monitored by TLC. After 12 hours of reaction, the reaction was almost complete. The reaction solution was taken out and concentrated in vacuo. The residue was dissolved with ethyl acetate (50 mL). The organic phase was washed with water (3×20 mL), and then dried with anhydrous Na2SO4. The organic phase was concentrated in vacuo to remove the solvent to obtain an active ester product (white solid, 460 mg, y=84%).

[0174] Take 300 mg of the active ester product obtained in the previous step, place it in a round-bottom flask with a magnetic stirrer, add 5 mL of THF to dissolve, and stir under ice bath. Dissolve NaHS xH2O (137 mg, 1.66 mmol, 2 eq) in water (less than 1 mL), slowly drip into the THF solution of the active ester under ice bath, monitor the reaction by TLC, take out the reaction solution after 5 minutes of reaction, concentrate it in vacuo, 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 it once with saturated brine, dry it with anhydrous sodium sulfate, combine the organic phases, concentrate it in vacuo to remove the solvent, use a small amount of DCM solvent residue, and load it onto a silica gel column, elute it with DCM containing 1% acetic acid, collect the eluate, remove most of the solvent under reduced pressure, add a large amount of PE, and a white powder solid precipitates. Filter, wash the precipitate with PE, and obtain ZYSD02 (white powder solid, 49 mg, y=21%).

[0175] 1 H NMR (300MHz, Methanol-d4) δ7.26(d,J=8.03Hz,2H),7.10(d,J=8.10Hz,2H),4.04(d,J=2.74Hz,2H),3.69(q,J=7.0 8Hz, 1H), 2.44 (d, J = 7.18Hz, 2H), 1.83 (hept, J = 13.54, 6.76Hz, 1H), 1.46 (d, J = 7.14Hz, 3H), 0.89 (d, J = 6.61Hz, 6H).

[0176] 13 C NMR (75MHz, Methanol-d4) δ175.56,168.55,140.18,138.77,128.96,126.98,45.64,44.73,40.58,30.20,21.41,17.61.

[0177] Example 16 Compound ZYSH01 (Ibuprofen)

[0178]

[0179] Ibuprofen (4mmol, 825mg, 1eq) and Lawesson's reagent (2.2mmol, 890mg, 0.55eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (10mL) was added as solvent with a syringe. The reaction was stirred at 80°C and monitored by TLC. After about 1 hour of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0180] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, cyclohexane was added, and a white solid was precipitated. Thiobuprofen sodium salt (white solid, 658 mg, y = 67%) was obtained by filtration.

[0181] 1 H NMR(400MHz, Methanol-d4)δ7.30–7.25(m,2H),7.07–6.97(m,2H),4.16(q,J=7.1Hz,1H),2.4 1(d,J=7.1Hz,2H),1.81(dp,J=13.5,6.8Hz,1H),1.40(d,J=7.1Hz,3H),0.88(d,J=6.6Hz,6H).

[0182] 13 C NMR (101MHz, Methanol-d4) δ142.27,140.36,129.66,128.36,60.80,46.11,31.50,22.73,20.42.

[0183] Example 17 Compound ZYSH02 (Indomethacin)

[0184]

[0185] 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 in sequence. The reaction was monitored by TLC. After stirring at room temperature for 2 h, the reaction was almost complete. The reaction solution was concentrated in vacuo, and the residue was dissolved with a small amount of DCM and loaded onto a silica gel column. Indomethacin active ester (white solid, 1.24 g, y = 91%) was obtained by elution with DCM.

[0186] Indomethacin active ester (1.24 g) was dissolved in THF (15 mL) and placed in an ice bath to cool. NaHS xH2O (68%) (546 mg, 2.73 mmol, 2 eq) was dissolved in water (less than 5 mL). NaHS xH2O aqueous solution was slowly added dropwise to the THF solution of indomethacin 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 TLC plate was used to determine that the reaction was complete, the reaction solution was concentrated in vacuo, the residue was dissolved with DCM and then loaded onto a silica gel column. The crude thioindomethacin product (light yellow solid) was obtained by eluting with DCM.

[0187] The crude thioindomethacin product was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. The product was placed in an ice bath to cool down. After cooling down, a MeOH solution of NaOH (0.8 eq) was added, and the product was stirred in an ice bath for 5 minutes. After the reaction solution was concentrated in a vacuum, a small amount of DCM was added to dissolve the residue. Anhydrous ether was added, and a white solid was precipitated. Thioindomethacin sodium salt (white solid, 854 mg, y = 72%) was obtained by filtration.

[0188] 1 H NMR (300MHz, Methanol-d4) δ7.74–7.67(m,2H),7.58–7.51(m,2H),7.13(d,J=2.5Hz,1H),6.95(d ,J=8.9Hz,1H),6.62(dd,J=9.0,2.6Hz,1H),4.04(s,2H),3.80(s,3H),2.26(s,3H),1.90(s,1H).

[0189] 13 C NMR(101MHz,Methanol-d4)δ180.47,167.57,154.63,140.44,134.56,132.15,132.08,130 .72,129.98,129.83,111.51,110.80,108.50,101.90,56.29,47.65,24.23,11.85,-15.68.

[0190] Example 18 Compound ZYSH03 (Ketoprofen)

[0191]

[0192] 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 in sequence. The reaction was monitored by TLC. After stirring for 2 h at room temperature in the dark, the reaction was almost complete. The reaction solution was concentrated in vacuo, and the residue was dissolved with a small amount of DCM and loaded onto a silica gel column. Ketoprofen active ester (white solid, 874 mg, y = 83%) was obtained by elution with DCM.

[0193] Ketoprofen active ester (800mg, 2.27mmol, 1eq) was dissolved in THF (15mL), placed in an ice bath to cool, NaHSxH2O (68%) (455mg, 4.55mmol, 2eq) was dissolved in water (less than 5mL), and NaHSxH2O aqueous solution was slowly added dropwise to the THF solution of ketoprofen ester in an ice bath, and the ice bath was removed after the addition was complete, and the mixture was stirred at room temperature. After TLC spot plate confirmed that the reaction was complete, the reaction solution was concentrated in a vacuum, and the residue was dissolved with DCM and then loaded onto a silica gel column, and the DCM wash-out obtained the thioketoprofen product (light yellow solid).

[0194] The thioketoprofen crude product is placed in a round-bottomed flask equipped with a magnetic stirrer, add an appropriate amount of DCM to dissolve, and it is placed in an ice bath to cool down. After cooling is completed, add the MeOH solution of NaOH (0.8eq), stir in an ice bath for 5 minutes. The reaction solution is concentrated in a vacuum, and cyclohexane is added. TLC determines purity, and constantly washes away the little assorted spots of polarity with DCM / cyclohexane. Spin-drying is dissolved in the tert-butyl alcohol, and lyophilized back obtains thioketoprofen sodium salt (light yellow foamy solid, 307mg, y=35%).

[0195] 1 H NMR (300MHz, Chloroform-d) δ7.83–7.70(m,4H),7.65–7.43(m,5H),4.25–3.89(m,1H),1.58(dd,J=19.6,7.1Hz,3H).

[0196] Example 19 Compound ZYSH04 (Gemfibrozil)

[0197]

[0198] Gemfibrozil and Lawesson's reagent (0.55 eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene was added as solvent with a syringe. The reaction was stirred at 90°C and monitored by TLC. After about 2 hours of reaction, the system changed from turbid to clear, indicating that the reaction was almost complete. The reaction solution was taken out, the toluene was dried by spin drying, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0199] The crude product obtained by column chromatography in the previous step is placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of MeOH is added to dissolve it. It is placed in an ice-salt bath to cool down. After cooling, a methanol solution of NaOH (0.8 eq) is added, and stirred in an ice-salt bath for 5 minutes. After the reaction solution is spin-dried, anhydrous ether is added, and a white solid will precipitate. It can be filtered to obtain thiogemfibrozil sodium salt (white solid).

[0200] 1 H NMR (400MHz, Methanol-d4) δ6.92(d,J=7.5Hz,1H),6.65(d,J=1.5Hz,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=3.4Hz,3H),1.21(s,6H).

[0201] 13 C NMR (101MHz, Methanol-d4) δ157.06,136.12,129.69,122.99,120.14,111.61,68.41,50.92,38.81,26.75,25.15,20.10,14.64.

[0202] Example 20 Compound ZYSH05 (probenecid)

[0203]

[0204] Probenecid (285 mg, 1 mmol, 1 eq) was added into a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, DCM (2 mL), oxalyl chloride (170 uL, 4 mmol, 2 eq), and DMF (2 drops) were added in sequence with a syringe. After stirring at room temperature for 1 h, the reaction solution was taken out and concentrated under vacuum to obtain a yellow oil (310 mg), which was dissolved in dry THF (3 mL).

[0205] Place the THF solution of probenecid chloride in an ice bath and cool it with stirring. Dissolve sodium hydrosulfide hydrate (136 mg, 2 mmol, 2 eq) in a small amount of water (less than 2 mL) and slowly add it dropwise to the THF solution of probenecid chloride. After the addition is complete, stir the reaction in an ice bath for 1 h, then transfer to room temperature for 1 h, and quench the reaction with 1M HCl. Extract the aqueous phase three times with ethyl acetate (3×20 mL). Combine the organic layers, dry them over anhydrous NaSO4, and concentrate them in vacuo to obtain a crude thioprobenecid product (yellow oil).

[0206] The crude product obtained in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of MeOH was added to dissolve it. It was then placed in an ice-salt bath to cool. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After MeOH was removed in vacuo, a small amount of DCM was added to the residue to dissolve it. Anhydrous ether was added to precipitate a bright yellow solid, which was filtered to obtain thioprobenecid sodium salt (bright yellow solid, 119 mg, y = 36%).

[0207] 1 H NMR (300MHz, Methanol-d4) δ8.26–8.18(m,2H),7.77–7.70(m,2H),3.10(d,J=7.6Hz,4H),1.55(h,J=7.4Hz,4H),0.88(t,J=7.4Hz,6H).

[0208] 13 C NMR (101MHz, Methanol-d4) δ212.67,149.16,141.93,129.60,127.23,51.31,23.11,11.43.

[0209] Example 21 Compound ZYSH06 (Telmisartan)

[0210]

[0211] Telmisartan (500 mg, 0.99 mmol) was placed in a three-necked flask, vacuumed to replace N2, DCM was added to dissolve it, oxalyl chloride (137 mg, 125 μL) was added dropwise under ice bath conditions, and then a catalytic amount of DMF was added dropwise, and then the mixture was allowed to react at room temperature for 1 h, and then dry hydrogen sulfide gas was introduced, and the reaction was monitored by TLC, DCM:MeOH 15:1, and then purified by column chromatography to obtain 300 mg (57%) of a yellow solid. 1H NMR(300MHz, Methanol-d4)δ8.09(s,1H),7.98(d,J=5.2Hz,1H),7.89(d,J=9.4Hz,2H),7.83–7.65(m,3H),7.57(d,J=6.4Hz,1H), 7.46(d,J=6.8Hz,3H),7.39(d,J=7.3Hz,3H),5.92(s,2H),4.04(s,3H),2.86(s,3H),1.95(q,J=7.6Hz,2H),1.15(t,J=7.4Hz,3H).

[0212] Example 22 Compound ZYSH21 (Diclofenac)

[0213]

[0214] Synthesis of sodium 2-[(2,6-dichlorophenyl)amino]-phenylthioacetate (ZYSH21). Sodium 2-[(2,6-dichlorophenyl)amino]-phenylacetate (504 mg, 1.58 mmol) and Lawesson's reagent (351 mg, 0.869 mmol) were added to a round-bottom flask and dissolved in 20 ml of dichloromethane. The reaction was allowed to react at room temperature for 2.5 hours. The reaction was basically complete as monitored by TLC. The dichloromethane was removed by vacuum concentration to obtain a 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, and allowed to stand at -20°C for 30 minutes. The solvent was removed by vacuum concentration, and the final product was recrystallized in ether as a white solid (301 mg, 60%). 1HNMR (300MHz, DMSO-d6): δ9.17(s,1H),7.46(d,J=8.0Hz,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=7.8Hz, 1H), 3.87 (s, 2H); 13CNMR (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.

[0215] Example 23 Compound ZYSH31 (Naproxen)

[0216]

[0217] Naproxen (4mmol, 920mg, 1eq) and Lawesson's reagent (2.2mmol, 890mg, 0.55eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (10mL) was added as solvent with a syringe. The reaction was stirred at 80°C and monitored by TLC. After about 1 hour of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, the toluene was spin-dried, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (dark pink oil) was obtained by eluting with DCM.

[0218] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice-salt bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was spin-dried, anhydrous ether was added, and a white solid was precipitated. It was filtered to obtain thionaproxen sodium salt (white solid, 800 mg, y = 74%).

[0219] 1 H NMR(500MHz, Methanol-d4)δ7.73–7.61(m,3H),7.52(dd,J=8.6,1.7Hz,1H),7.15(d,J=2.5Hz ,1H),7.06(dd,J=8.9,2.5Hz,1H),4.32(q,J=7.1Hz,1H),3.88(s,3H),1.50(d,J=7.1Hz,3H).

[0220] 13 C NMR (126MHz, Methanol-d4) δ223.91,158.64,140.29,134.84,130.49,130.16,128.06,127.35,126.64,119.28,106.57,61.10,55.70,20.41.

[0221] (ESI): m / z calculated for C 14 H 13 Na O2 S requires 291.04262for[M+Na] + ,found291.04247.

[0222] Example 24 Synthesis of Compound ZYSH07:

[0223]

[0224] Flurbiprofen (4mmol, 976mg, 1eq) and Lawesson's reagent (2.2mmol, 890mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (20mL) was added as solvent with a syringe. The reaction was stirred at 95°C and monitored by TLC. After about 30 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, and the residue was dissolved in DCM, loaded onto a silica gel column, and eluted with DCM to obtain a crude product (dark pink oil).

[0225] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. It was filtered to obtain thioflurbiprofen sodium salt (white solid, 555 mg, y = 49%).

[0226] 1 H NMR (300MHz, Methanol-d4) δ7.52(dt,J=8.2,1.5Hz,2H),7.46–7.31(m,4H),7.29–7.19(m,2H),4.24(q,J=7.1Hz,1H),1.47(d,J=7.1Hz,3H).

[0227] 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.

[0228] Example 25 Synthesis of compound ZYSH08:

[0229]

[0230] Oxaprozin (3mmol, 879mg, 1eq) and Lawesson's reagent (1.8mmol, 728mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (15mL) was added as solvent using a syringe. The reaction was stirred at 90°C and monitored by TLC. After about 30 minutes of reaction, the system changed from turbid to clear, indicating that the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0231] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Thioxaprozin sodium salt (white solid, 470 mg, y = 47%) was obtained by filtration.

[0232] 1 H NMR (300MHz, Methanol-d4) δ7.45 (dddd, J=56.6, 7.8, 4.8, 2.1Hz, 10H), 3.18 (s, 4H).

[0233] 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.

[0234] Example 26 Synthesis of Compound ZYSA12:

[0235]

[0236] 3-Indolepropionic acid (3mmol, 567mg, 1eq) and Lawesson's reagent (1.8mmol, 728mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (15mL) was added as solvent with a syringe. The reaction was stirred at 100°C and monitored by TLC. After about 30 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, and the residue was dissolved in DCM, loaded onto a silica gel column, and eluted with DCM to obtain a crude product (pink oil).

[0237] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium 3-indolethiopropionic acid (white solid, 438 mg, y = 64%).

[0238] 1 H NMR (300MHz, Methanol-d4) δ7.57(dt,J=7.7,1.0Hz,1H),7.29(dt,J=8.1,1.0Hz,1H),7.08–6.93(m,3H),3.11–2.96(m,4H).

[0239] 13 C NMR (75MHz, Methanol-d4) δ223.45,138.06,128.74,122.55,122.03,119.37,119.23,116.05,111.99,53.02,24.54.

[0240] Example 27 Synthesis of Compound ZYSA13:

[0241]

[0242] Add p-chlorophenoxyisobutyric acid (4mmol, 858mg, 1eq) and Lawesson's reagent (2.4mmol, 970mg, 0.6eq) to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, add toluene (20mL) as solvent with a syringe, stir at 100°C to react, monitor the reaction by TLC, and after about 20 minutes of reaction, the system changes from turbid to clear, and the reaction is almost complete. Take out the reaction solution, remove toluene in vacuum, dissolve the residue in DCM, load it onto a silica gel column, and elute with DCM to obtain a crude product (pink oil).

[0243] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium thio-p-chlorophenoxyisobutyric acid (white solid, 612 mg, y = 60%).

[0244] 1H NMR(300MHz,Methanol-d4)δ7.12(d,J=9.0Hz,2H),6.86(d,J=9.0Hz,2H),1.61(s,6H)

[0245] 13 C NMR (75MHz, Methanol-d4) δ223.38,156.36,129.37,126.33,121.40,88.18,27.92.

[0246] Example 28 Synthesis of Compound ZYSB04:

[0247]

[0248] Eicosanoic acid (2mmol, 625mg, 1eq) and Lawesson's reagent (1.2mmol, 242mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (10mL) was added as solvent with a syringe. The reaction was stirred at 95°C and monitored by TLC. After about 20 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, and the residue was dissolved in DCM, loaded onto a silica gel column, and eluted with DCM to obtain a crude product (pink oil).

[0249] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium thioeicosanoic acid salt (white solid, 428 mg, y = 61%).

[0250] 1H NMR (300MHz, DMSO-d6) δ2.29 (t, J = 7.6Hz, 2H), 1.43 (s, 2H), 1.23 (s, 32H), 0.85 (s, 3H).

[0251] 13C NMR (75MHz, DMSO-d6) δ217.09,51.30,31.41,29.18,28.85,27.30,22.19,13.95.

[0252] Example 29 Synthesis of Compound ZYSB05:

[0253]

[0254] Hexadecanoic acid (3mmol, 769mg, 1eq) and Lawesson's reagent (1.8mmol, 728mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (15mL) was added as solvent with a syringe. The reaction was stirred at 95°C and monitored by TLC. After about 20 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0255] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium thiohexadecanoate (white solid, 675 mg, y = 76%).

[0256] 1 H NMR (300MHz, DMSO-d6) δ2.30 (t, J = 7.5Hz, 2H), 1.48–1.39 (m, 2H), 1.23 (s, 24H), 0.89–0.81 (m, 3H).

[0257] 13 C NMR (75MHz, DMSO-d6) δ216.68,51.34,31.35,29.13,28.97,28.77,27.26,22.15,13.98.

[0258] Example 30 Synthesis of Compound ZYSB06:

[0259]

[0260] Tetradecanoic acid (3mmol, 685mg, 1eq) and Lawesson's reagent (1.8mmol, 728mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (15mL) was added as solvent with a syringe. The reaction was stirred at 95°C and monitored by TLC. After about 15 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0261] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium thiotetradecanoic acid salt (white solid, 497 mg, y = 62%).

[0262] 1 H NMR (300MHz, Methanol-d4) δ2.64–2.58(m,2H),1.63(t,J=7.4Hz,2H),1.28(s,20H),0.95–0.84(m,3H).

[0263] 13 C NMR (75MHz, Methanol-d4) δ 224.09, 52.32, 33.09, 30.79, 30.50, 30.42, 29.00, 23.75, 14.48. Example 31 Synthesis of Compound ZYSB07:

[0264]

[0265] Dodecanoic acid (3mmol, 600mg, 1eq) and Lawesson's reagent (1.8mmol, 728mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (15mL) was added as solvent with a syringe. The reaction was stirred at 95°C and monitored by TLC. After about 15 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, and the residue was dissolved in DCM, loaded onto a silica gel column, and eluted with DCM to obtain a crude product (pink oil).

[0266] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. Filtration can obtain sodium thiododecanoic acid salt (white solid, 411 mg, y = 57%).

[0267] 1 H NMR (400MHz, DMSO-d6) δ2.33–2.27(m,2H),1.44(p,J=7.3Hz,2H),1.23(s,16H),0.87–0.83(m,3H).

[0268] 13C NMR (75MHz, DMSO-d6) δ 51.26, 31.36, 29.15, 28.95, 28.78, 27.25, 22.15, 13.99. Example 32 Synthesis of Compound ZYSB08:

[0269]

[0270] Adamantanecarboxylic acid (4mmol, 720mg, 1eq) and Lawesson's reagent (2.4mmol, 970mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (20mL) was added as solvent with a syringe. The reaction was stirred at 90°C and monitored by TLC. After about 10 minutes of reaction, the system changed from turbid to clear, indicating that the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, the residue was dissolved in DCM, and the sample was loaded onto a silica gel column. The crude product (pink oil) was obtained by eluting with DCM.

[0271] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added, and it was stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. It was filtered to obtain sodium thioadamantanecarboxylate (white solid, 525 mg, y = 60%).

[0272] 1 H NMR (300MHz, Methanol-d4) δ1.99–1.92 (m, 9H), 1.71 (t, J = 2.9Hz, 6H).

[0273] 13 C NMR (75MHz, Methanol-d4) δ229.92,51.23,42.43,38.05,30.40.

[0274] Example 33 Synthesis of Compound ZYSB09:

[0275]

[0276] Cyanocyanic acid (4mmol, 568mg, 1eq) and Lawesson's reagent (2.4mmol, 970mg, 0.6eq) were added to a two-necked round-bottom flask (oven dried) with a magnetic stirrer. After replacing N2, toluene (20mL) was added as solvent with a syringe. The reaction was stirred at 90°C and monitored by TLC. After about 10 minutes of reaction, the system changed from turbid to clear, and the reaction was almost complete. The reaction solution was taken out, toluene was removed in vacuo, and the residue was dissolved in DCM, loaded onto a silica gel column, and eluted with DCM to obtain a crude product (pink oil).

[0277] The crude product obtained by column chromatography in the previous step was placed in a round-bottom flask with a magnetic stirrer, and an appropriate amount of DCM was added to dissolve it. It was placed in an ice bath to cool down. After cooling, a MeOH solution of NaOH (0.8 eq) was added and stirred in an ice-salt bath for 5 minutes. After the reaction solution was concentrated in a vacuum, anhydrous ether was added, and a white solid was precipitated. It was filtered to obtain sodium thiocyanate (white solid, 481 mg, y = 66%).

[0278] 1 H NMR (300MHz, Methanol-d4) δ2.17 (s, 1H), 1.17 (d, J = 14.5Hz, 12H).

[0279] 13 C NMR (75MHz, Methanol-d4) δ218.04,52.65,32.03,24.27,17.13.

[0280] Example 34 Compound ZYSC03

[0281]

[0282] Referring to the synthesis method of target ZYSA01, p-methoxynaphthoic acid and Lawesson's reagent were used as raw materials to synthesize crude thio-p-methoxynaphthoic acid, which was then converted into sodium salt to obtain sodium thio-p-methoxynaphthoate, a light yellow solid.

[0283] 1 H NMR (300MHz, Methanol-d4) δ8.61(s,1H),8.19(dd,J=8.6,1.8Hz,1H),7.82(d,J=9.0Hz,1 H),7.68(d,J=8.6Hz,1H),7.23(d,J=2.5Hz,1H),7.12(dd,J=9.0,2.5Hz,1H),3.91(s,3H).

[0284] 13C NMR (75MHz, 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.

[0285] Performance data:

[0286] Example 35 Determination of the half-life of hydrogen sulfide release

[0287] 1. Detection of PLE hydrolysis of ZYSA series thiocarboxylic acids and the formation of corresponding carboxylic acids by HPLC

[0288] Add 4.5 mL of porcine liver esterase (PLE) (3 U / mL) in PBS buffer to a 20 mL reaction bottle, then add 0.5 mL of ZYSA in PBS buffer (5 mM), and incubate in a 37°C water bath. Pipette an equal amount of 200 μL of solution at each time point and transfer it to a 1.5 mL centrifuge tube containing 200 μL of chromatographic acetonitrile. After transferring to a -20°C refrigerator for rapid freezing, centrifuge for 10 min (13.0 x 1000 r / min), and aspirate the supernatant for HPLC detection.

[0289] 2. Determination of hydrogen sulfide release from ZYSA series thiocarboxylic acids by methylene blue method

[0290] ZYSA was dissolved in 5 mL of PBS buffer solution with a pH of 7.4 (final concentration of 500 μM) and incubated with 3 U / mL PLE. An equal amount of 200 μL of solution was taken at each time point and transferred to a 1.5 mL centrifuge tube containing 200 μL of zinc acetate (1%, w / v). The centrifuge tube was then centrifuged for 10 minutes (13.0 x 1000 r / min). The supernatant was removed 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 tube. After being transferred to a 37°C metal bath and incubated for 30 minutes, the absorbance at 740 m was detected by a UV absorption spectrophotometer. The hydrogen sulfide concentration was calculated based on the sodium hydrosulfide standard curve.

[0291] The results are shown in Table 1. By the methylene blue method, the compounds of the present invention can detect the generation of relevant hydrogen sulfide. Under the conditions of esterase, they can basically release hydrogen sulfide at a certain rate. The specific released t 1 / 2 As shown in Table 1.

[0292] Table 1 Structure, nomenclature and half-life of hydrogen sulfide release of thiocarboxylic acids in the examples of this patent

[0293]

[0294]

[0295] Example 36 Effect data of compound ZYSA01 prepared in Example 1

[0296] 1. Determination of infarct area

[0297] The mouse brain was 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. The infarcted area was not stained, and the normal area was stained red ( Figure 1 A). 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 area / total area ( Figure 1 B). The results showed that ZYSA01 was significantly better than benzoic acid.

[0298] 2. Determination of serum LDH level and brain tissue LDA level

[0299] After blood collection, serum was collected by centrifugation at 3000g for 10 minutes at 4°C. For the determination method of LDH, please refer to the instructions of the kit (solarbio, Beijing). Brain tissue was taken to determine the level of MDA in the tissue. For the determination method, please refer to the instructions of the MDA kit (solarbio, Beijing). The LDH level reflects the cell membrane damage caused by ischemia-reperfusion injury, and the MDA level reflects the peroxidation of membrane lipids 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.

[0300] Example 37 Compound ZYSH06 (Telmisartan) Activity Data

[0301] 1. Verification of cerebral infarction activity

[0302] Mouse middle cerebral artery occlusion model 25-30g C57 / 6J mice were randomly divided into two groups: a control group and a drug group. The dose of telmisartan was 0.5×10 -4mol / kg The rest of the groups were subjected to the middle cerebral artery occlusion model. Anesthesia was induced with 3% isoflurane, and anesthesia was maintained with 0.6% isoflurane after anesthesia. 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 at the distal end, and a suture (Beijing Xinong Biotechnology Co., Ltd.) was inserted and passed through the ICA until the cerebral artery (MCA) was blocked in the middle. The drug was administered by gavage during infarction, and 1 hour and 20 minutes after infarction, we removed the suture and reperfused. Other treatments were the same as above. Blood was collected 24 hours later, and the mice were killed by cervical dislocation, the skull was opened, and brain tissue was collected for subsequent experiments.

[0303] 2. Infarct size

[0304] The mouse brain was 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-triphenyltetrazolium chloride (TTC) solution at 37°C for 30 minutes and fixed in 4% paraformaldehyde (PFA) solution. The infarct area was not stained, and the normal area was stained red. Each pair of slices was then photographed, and the area of ​​cerebral infarction was calculated using ImageJ. The formula is as follows: Infarct volume = sum of infarct areas / total area. The results of the study showed that both positive drugs and prodrugs can significantly reduce the infarct area, and showed a dose-dependent effect. The efficacy of ZYSA06 was significantly better than that of positive drugs, among which ZYSA06 showed greater superiority in reducing the infarct area. Figure 2 A, 2B.

[0305] 3. Serum LDH level and brain tissue MDA level

[0306] After blood collection, serum was collected by centrifugation at 3000g for 10 minutes at 4°C. The method for determining LDH is shown in the instructions of the kit (solarbio, Beijing). Brain tissue was taken to determine the level of MDA in the tissue. The determination method is shown in the instructions of the MDA kit (solarbio, Beijing). The LDH level reflects the cell membrane damage caused by ischemia-reperfusion injury, and the MDA level reflects the peroxidation of membrane lipids caused by ischemia-reperfusion injury. The study found that ZYSA06 showed greater potential in the MCAO model, 0.25×10 - 4 mol / kg ZYSA09 can significantly reduce the MDA level, and the telmisartan group showed a significant difference *P<0.05 Figure 2 C,2D.

[0307] Example 38 Compound ZYSH21 (Diclofenac) Activity Data

[0308] 1. ZYSH21 releases the original drug DCF in rats

[0309] Five rats were fasted for 10 h before administration, with free access to water. They were given 10.50 mg / kg ZYSH21 solution, and blood samples were collected at (0 h) and 0.033, 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 6, and 8 h after administration. A 5 μL IS solution (1000 ng / ml naproxen) aliquot was added to a 50 μL plasma sample in a centrifuge tube, and after vortex mixing for 10 seconds, 200 μL methanol was added to the mixture to precipitate the protein. The sample was vortex mixed for 1 minute and centrifuged at 12000 rpm for 5 min at 4 ° C. Finally, the supernatant was measured by triple quadrupole liquid chromatography-mass spectrometry. ( Figure 3 )

[0310] 2. Anti-inflammatory effect of ZYSH21

[0311] Wistar rats were divided into two groups, 8 rats in each group: ZYSH21 (10 mg / kg / day), DCF (10 mg / kg / day), immune-grade natural type II collagen was dissolved in 0.05 mol / l acetic acid to make a 2 mg / ml solution, and type II collagen was homogenized with Freund's incomplete adjuvant (1:1) at 4°C to prepare an emulsion with a final concentration of CII of 1 mg / ml. The emulsified mixture was fully emulsified in an ice bath until it did not diffuse when added to water. The emulsified mixture was injected subcutaneously at the base of the tail at 0.2 ml / rat, i.e. 200 μg CII / rat. After 7 days, 0.1 ml / rat, i.e. 100 μg CII / rat, was injected subcutaneously at multiple points on the back. After 14 days, the swelling of the toes reached its peak. The drug was administered continuously for 28 days, and the changes in the degree of swelling of the toes were recorded. The swelling rate of the toes was evaluated 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 ).

[0312] 3. Gastrointestinal safety of ZYSH21

[0313] Wistar rats were divided into ZYSH21 (10mg / kg / day, 5mg / kg / day) and DCF (10mg / kg / day, 5mg / kg / day), 8 rats in each group, and the drugs were continuously administered for 28 days. Among them, the ulcer area of ​​ZYSH21 (10mg / kg / day, 5mg / kg / day) was extremely low, while DCF (10mg / kg / day, 5mg / kg / day) had obvious ulcer spots ( Figure 5 ).

[0314] 4. ZYSH21 acute liver injury

[0315] C57BL / 6JNifdc mice were fasted for 10 hours before administration and were allowed to drink water freely. The mice were randomly divided into three groups: saline control group, DCF group, and ZYSH21 group. The DCF dose was 150 mg / kg, and ZYSH21 was administered in equal moles, with 6 mice in each group. The administration method was intraperitoneal injection, and the administration duration was 24 hours. Blood was collected from the eyeballs at 24 hours and the mice were killed by spinal dislocation. The serum alanine transaminase (ALT) and aspartate aminotransferase (AST) levels were measured ( Figure 6 ).

[0316] 5. ZYSH21 kidney damage

[0317] Wistar rats were divided into two groups, 8 in each group: ZYSH21 (10 mg / kg / day), DCF (10 mg / kg / day), and administered for 28 consecutive days. The activity of uric acid (UA), an indicator of renal damage, decreased significantly and remained within the normal level (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) ( Figure 7 ).

[0318] Example 39 ZYSH31 (naproxen) activity data

[0319] 1. PLE enzymatic hydrolysis of ZYSH31 releases the original drug NPX

[0320] Take a 5 mL sample bottle, add 50 μL 10 mM ZYSH31 solution, 4950 μL PBS buffer and 1 mg PLE to the sample bottle, seal the sample bottle with film, incubate the sample bottle in a 37 ° C water bath, take 200 μL of solution at regular intervals, filter and place in a 1.5 mL injection bottle, and use appropriate HPLC mobile phase and elution to analyze the components in the above solution. 1 / 2 208.4±2.1min( Figure 8 ).

[0321] 2. ZYSH31 releases the original drug NPX in rats

[0322] Five rats were fasted for 10 h before administration and were allowed to drink water freely. They were given 10.60 mg / kg ZYSH31 solution, and blood samples were collected at (0 h) and 0.033, 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 6, and 8 h after administration. A 5 μL IS solution (1000 ng / ml diclofenac) aliquot was added to 50 μL plasma sample in a centrifuge tube, and after vortex mixing for 10 seconds, 200 μL methanol was added to the mixture to precipitate protein. The sample was vortex mixed for 1 minute and centrifuged at 12000 r·min at 4 °C. -1 Centrifuge for 5 minutes, and finally use triple quadrupole liquid chromatography-mass spectrometry to measure the supernatant ( Fig. 9 ).

[0323] 3. Anti-inflammatory effect of ZYSH31

[0324] Wistar rats were divided into two groups, 6 rats in each group: ZYSH31 (10 mg / kg / day), NPX (10 mg / kg / day), immune grade natural type II collagen was dissolved in 0.05 mol / l acetic acid to make a 2 mg / ml solution, and type II collagen was homogenized with Freund's incomplete adjuvant (1:1) at 4°C to prepare an emulsion, with a final concentration of CII of 1 mg / ml. The emulsified mixture was fully emulsified in an ice bath, and the emulsified mixture was injected subcutaneously at the base of the tail at a rate of 0.2 ml / rat, i.e. 200 μg CII / rat. After 7 days, 0.1 ml / rat, i.e. 100 μg CII / rat, was injected subcutaneously at multiple points on the back. After 14 days, the swelling of the toes reached its peak, and the drug was administered continuously for 28 days, and the changes in the degree of swelling of the toes were recorded. The swelling rate of the toes was evaluated from day 0 to day 36. Compared with the normal group: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.( Fig.10 ).

[0325] 4. Gastrointestinal safety of ZYSH31

[0326] Wistar rats were divided into ZYSH31 (10mg / kg / day, 5mg / kg / day) and NPX (10mg / kg / day, 5mg / kg / day), 6 rats in each group, and the drugs were given for 28 consecutive days. Among them, the ulcer area of ​​ZYSH31 (10mg / kg / day, 5mg / kg / day) was extremely low, while NPX (10mg / kg / day, 5mg / kg / day) had obvious ulcer spots ( Fig.11 A).

[0327] 5. ZYSH31 acute liver injury

[0328] C57BL / 6JNifdc mice were fasted for 10 hours before administration and were allowed to drink water freely. The mice were randomly divided into three groups: saline control group, NPX group, and ZYSH31 group. The NPX dose was 150 mg / kg, and the ZYSH31 was administered in equal moles, with 6 mice in each group. The administration method was intraperitoneal injection, and the administration duration was 24 hours. After 24 hours, blood was collected from the eyeballs and the mice were killed by spinal dislocation. The serum alanine transaminase (ALT) and aspartate aminotransferase (AST) levels were measured ( Fig.11 B).

[0329] 6. ZYSH31 kidney damage

[0330] Wistar rats were divided into two groups, 6 in each group: ZYSH31 (10 mg / kg / day), NPX (10 mg / kg / day), administered for 28 consecutive days), the activity of albumin, an indicator of renal damage, increased abnormally and remained within the normal level (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) ( Fig.12 ).

[0331] Example 40 Water Environment Stability

[0332] 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 and placed at room temperature to observe their stability. 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 had good stability and no obvious decomposition was observed within 144 hours.

Claims

1. A hydrogen sulfide prodrug, characterized in that The invention is to sulfide the existing drugs with carboxylic acid structure to form thiocarboxylic acid structure, which can release H2S under enzyme catalysis.

2. The hydrogen sulfide prodrug according to claim 1, characterized in that The enzyme is an enzyme that can decompose the thiocarboxylic acid structure into a carboxylic acid structure and release H2S; preferably, it is an esterase.

3. The hydrogen sulfide prodrug according to claim 1, characterized in that The existing drugs with a carboxylic acid structure are drugs for treating cardiovascular and cerebrovascular diseases, anti-inflammatory drugs, antipyretic and analgesic drugs, rheumatic or rheumatoid arthritis drugs, anti-gout drugs, blood pressure lowering drugs, blood lipid lowering drugs or drugs that reduce GSH consumption or liver toxicity caused by mitochondrial damage; preferably, the following types of drugs with a -COOH structure: clofibric acid lipid regulating drugs, sartan drugs, sulfonamide drugs, indole drugs, and non-steroidal anti-inflammatory drugs.

4. A hydrogen sulfide prodrug, characterized in that: The invention relates to sulfiding the existing sartan drugs with -COOH structure to form thiocarboxylic acid structure, which can release H2S under enzyme catalysis. The sartan drugs are sartan drugs with -COOH structure, preferably valsartan, eprosartan and telmisartan.

5. A hydrogen sulfide prodrug, characterized in that: It has a structure shown in the following formula (I), formula (II), formula (III) or (IV): Among them, in the compound of formula (I): Z + ChoiceNa + , K + , H + ; A is selected from C6-C 20 Aryl or C3-C 20 Heteroaryl; preferably C6-C 10 Aryl or C3-C 10 Heteroaryl; more preferably, the following groups: phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, purinyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, indolyl, benzotriazolyl, benzothienyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl or carbazolyl; L1 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl, -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-, -O-(CH2) n -、-O-(CH2) n -O-; n is selected from 0 or an integer from 1 to 8; R1 and R2 are independently selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -SO2NR3R4-, C6-C 10 Aryl or C3-C 10 heteroaryl; R3, R4, R5, R6 are independently selected from H or C1-C6 alkyl; Among them, in the compound of formula (II): Z + ChoiceNa + , K + , H + ; R' is selected from the following substituted or unsubstituted groups: C1-C 30 Alkyl, C3-C 10 Cycloalkyl, C8-C 16 Bridged cycloalkane, C6-C 16 Spirocycloalkane; the substituent is one or more, and the substituent is selected from H, halogen, -OH, CN, NO2, C1-C6 alkyl; Among them, in the compound of formula (III): Z + ChoiceNa + , K + , H + ; L2 is selected from a covalent bond, -(CH2) n - or C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4; R7 is selected from H, C1-C6 alkyl; 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 aryl or substituted or unsubstituted C3-C 10 Heteroaryl, substituents selected from H, halogen, -OH, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl; R9 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy; Among them, in the compound of formula (IV): Z + ChoiceNa + , K + , H + ; L3 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl; n is selected from 0, 1, 2, 3 or 4; X is selected from -NH- or -C(=O)-; R a , R b Independently selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy.

6. The hydrogen sulfide prodrug according to claim 5, characterized in that In the compound of formula (I): A is selected from C6-C 10 Aryl or C3-C 10 Heteroaryl; A is preferably a phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, purinyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, indolyl, benzotriazolyl, benzothienyl, benzofuranyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, tetrazolyl, triazinyl or carbazolyl.

7. The hydrogen sulfide prodrug according to claim 5, characterized in that In the compound of formula (I): L1 is selected from a covalent bond, -(CH2) n -, C2-C6 branched alkyl, -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-; preferably, L1 is selected from a covalent bond, -(CH2) n -、-CH(CH3)-、 -O-(CH2) n -C(CH3)2-, -R5C(=O)NHR6-.

8. The hydrogen sulfide prodrug according to claim 5, characterized in that In the compound of formula (I): n is selected from 0, 1, 2, 3 or 4.

9. The hydrogen sulfide prodrug according to claim 5, characterized in that In the compound of formula (I): R1 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C6-C 10 Aryl or C3-C 10 Heteroaryl; R1 is preferably the following groups: H, halogen, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl.

10. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (I), R2 is selected from H, halogen, CN, NO2, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -SO2NR3R4-, C6-C 10 Aryl or C3-C 10 Heteroaryl; R2 is preferably the following groups: H, halogen, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, -CH2Cl, -CF3, -CH2CF3, -CH2CCl3, -SO2NR3R4-, phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl.

11. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (I): R3, R4, R5, and R6 are independently selected from C1-C6 alkyl; preferably, R3, R4, R5, and R6 are independently selected from methyl, ethyl, propyl, isopropyl, butyl, and tert-butyl.

12. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (II): R' is selected from the following substituted or unsubstituted groups: C2-C6 branched alkyl, C1-C 20 Straight chain alkyl, C3-C6 cycloalkyl, C8-C 16 The substituents are one or more, and the substituents are selected from H, halogen, -OH, CN, NO2, and C1-C4 alkyl.

13. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (III): L2 is selected from -(CH2) n -; n is selected from 0, 1, 2 or 3.

14. The hydrogen sulfide prodrug according to claim 5, 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.

15. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (III): R8 is selected from H or -C(=O)-R 10 ; R 10 Selected from the following substituted or unsubstituted groups: phenyl, naphthyl, anthracenyl, oxazolyl, pyrrolyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyrimidinyl, quinolyl; the substituents are selected from H, halogen, -OH, CN, NO2, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl.

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

17. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (IV): L3 is selected from a covalent bond, -(CH2) n -、-CH(CH3)-、 n is selected from 0, 1, 2 or 3.

18. The hydrogen sulfide prodrug according to claim 5, characterized in that: In the compound of formula (IV): R a , R b Independently selected from H, halogen, CN, NO2, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

19. A hydrogen sulfide prodrug having the following structure:

20. A pharmaceutical composition comprising the hydrogen sulfide prodrug according to any one of claims 1 to 19 and a pharmaceutically acceptable carrier.

21. Use of the hydrogen sulfide prodrug according to any one of claims 1 to 19 in the preparation of a drug for treating cardiovascular and cerebrovascular diseases, anti-inflammatory, antipyretic and analgesic, rheumatic or rheumatoid arthritis, anti-gout, lowering blood pressure, lowering blood lipids, or reducing drug consumption of GSH or liver toxicity caused by mitochondrial damage.

22. A method for forming H2S, comprising contacting the hydrogen sulfide prodrug according to any one of claims 1 to 19 with an enzyme, wherein the thiocarboxylic acid structure forms a carboxylic acid structure under enzyme catalysis and releases H2S.

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