GSTO1 inhibitor and application thereof

Abstract

The invention belongs to the field of chemical synthesis, and relates to a GSTO1 inhibitor and application thereof, and application of the compound in medicines for treating or prophylactically treating diseases related to GSTO1, in particular to a compound shown as a formula I, or an isomer thereof, or pharmaceutically acceptable salt thereof and application thereof, and substituent groups in the formula I are as defined in the specification.

Description

Technical Field

[0001] This invention belongs to the field of chemical synthesis, specifically relating to a GSTO1 inhibitor and its uses. Background Technology

[0002] Glutathione thiotransferases (GSTs) are key enzymes in the glutathione binding reaction. GSTs participate in catalyzing the conjugation of glutathione with various endogenous and exogenous electrophilic compounds (including drugs, industrial intermediates, pesticides, herbicides, environmental pollutants, and carcinogens), playing an important role in cellular detoxification. They also play crucial roles in the synthesis and metabolism of endogenous compounds, redox homeostasis, and cell signal transduction. GSTs are classified into seven classes, with their Roman numerals A, M, P, S, T, O, and Z. GSTOs belong to one type of atypical cytoplasmic glutathione thiotransferase, and two isoforms, GSTO1 and GSTO2, have been identified in humans. GSTOs have low sequence homology with other GSTs and possess an atypical catalytic cysteine ​​residue, Cys32, at their active site.

[0003] Existing covalent inhibitors targeting the Cys32 active site of GSTO1 are mainly designed based on chloroacetamide warheads. For example, a series of compounds such as KT53 developed by Benjamin F. Cravatt et al. are covalent inhibitors against GSTO1 designed and optimized based on chloroacetamide warheads. However, KT53 has poor stability and undergoes intramolecular substitution reactions at room temperature, leading to loss of activity [J. Am. Chem. Soc. 2011, 133, 41, 16605-16616]. In addition, Nouri Neamati et al. developed C1-27 (CAS: 568544-03-6), a GSTO1 inhibitor based on chloroacetamide warheads, which has good GSTO1 inhibitory activity, but C1-27 has high cytotoxicity [J. Med. Chem. 2020, 63, 6, 2894-2914]. Acrylamide is the most common warhead among marketed covalent drugs. Only Nouri Neamati et al. have reported GSTO1 inhibitors of this class, but the compounds have poor drug-likeness, lack verification of target occupancy at the cellular level, and lack research on the mechanism of action [J.Med.Chem.2019,62,6,3068-3087]. Summary of the Invention

[0004] Purpose of the invention: The purpose of this invention is to overcome the shortcomings of the existing technology and provide a GSTO1 inhibitor and its application.

[0005] To achieve the above objectives, the technical solution provided by the present invention is as follows:

[0006] A compound of Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] Among them, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -C(R1)-, -CH2-, -CH-, -NR4-, -O- or -S-, and L3 is selected from -C(R3)-, -CH(R3)-, -CH2-;

[0009] R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C groups. 1-4 Alkyl, wherein the C 1-4 The alkyl group is substituted by one or more hydrogens or halogens, wherein the chiral carbon attached to the -OH group is in the R configuration;

[0010] R1 and R2 can form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 can form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is substituted by one or more hydrogens or halogens.

[0011] R4 is selected from hydrogen, -OH, or C. 1-4 alkyl;

[0012] n = 1 or 2;

[0013] L4, L5, L6, L7, and L8 are each independently selected from -C-, -C(R5)-, -N-, -O-, and -S-;

[0014] R5 is selected from hydrogen, halogen, -OH, cyano, carboxyl, or C. 1-4 alkyl;

[0015] R6 is selected from hydrogen, halogens, and C. 14 Alkyl or hydroxyl;

[0016] R8 is selected from halogen, cyano, and C. 1-4 Alkoxy or C 14 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more hydrogens, halogens, or C atoms. 1-4 Alkyl substitution;

[0017] R9 is selected from hydrogen, halogens, and C. 1-4 Alkyl or dimethylaminosulfonyl;

[0018] R7, R 10 Selected from hydrogen, halogen or C 1-4 alkyl;

[0019] R 11 It is selected from hydrogen, halogen or cyano.

[0020] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, is described by formula I-1:

[0021]

[0022] Among them, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -C(R1)-, -CH2-, -CH-, -NR4-, -O- or -S-, and L3 is selected from -C(R3)-, -CH(R3)-, -CH2-;

[0023] R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C groups. 1-4 Alkyl, wherein the C 1-4 The alkyl group is substituted by one or more hydrogens or halogens, wherein the chiral carbon attached to the -OH group is in the R configuration;

[0024] R1 and R2 can form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 can form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is substituted by one or more hydrogens or halogens.

[0025] R4 is selected from hydrogen, -OH, or C1-4 alkyl;

[0026] n = 1 or 2;

[0027] L5, L7, and L8 are each independently selected from -C(R5)-, -N-, -O-, and -S-, respectively, with R5 selected from hydrogen.

[0028] R6 is selected from hydrogen, halogens, and C. 1-4 Alkyl or hydroxyl;

[0029] R8 is selected from halogen, cyano, and C. 1-4 Alkoxy or C 1-4 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more hydrogens, halogens, or C atoms. 1-4 Alkyl substitution;

[0030] R9 is selected from hydrogen, halogens, and C. 1-4 Alkyl or dimethylaminosulfonyl;

[0031] R 11 It is selected from hydrogen, halogen or cyano.

[0032] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, Selected from:

[0033]

[0034] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, is described by formula I-2:

[0035]

[0036] When n=1, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -CH2-, -CH- or -S-, and L3 is selected from -CH(R3)- or -CH2-.

[0037] R1, R2, and R3 are independently selected from hydrogen, halogens, or -OH, wherein the chiral carbon attached to -OH is in the R configuration; or R1 and R2 form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is optionally substituted by one or more hydrogens or halogens.

[0038] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the 6-membered aromatic ring is a benzene ring.

[0039] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, Selected from:

[0040]

[0041] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, when n=2, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -CH2-, -NR4-, -O-, or -S-, and L3 is selected from -CH(R3)- or -CH2-.

[0042] R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C atoms. 1-4 Alkyl, wherein the C 1-4 The alkyl group is optionally substituted with one or more hydrogens or halogens, wherein the chiral carbon attached to the -OH group is in the R configuration.

[0043] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, Selected from:

[0044]

[0045] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R8 is selected from halogens, cyano groups, C... 1-4 Alkoxy or C 1-4 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more of the following hydrogens, halogens, or C atoms. 1-4Alkyl substitution; R6 is selected from hydrogen or hydroxyl; R9 is selected from hydrogen, halogen or dimethylaminosulfonyl.

[0046] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, Selected from:

[0047]

[0048] The compound, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the compound is selected from any of the following compounds:

[0049]

[0050] A pharmaceutical composition comprising the compound, its stereoisomer or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0051] The use of the compound, or its solvates, metabolites, cocrystals, prodrugs, or combinations thereof, in a medicament for the treatment or prophylactic treatment of diseases regulated by GSTO1 inhibitors.

[0052] The diseases mentioned include cancer, inflammation, or age-related macular degeneration.

[0053] The cancers include breast cancer, ovarian cancer, cervical cancer, pancreatic cancer, colorectal cancer, esophageal squamous cell carcinoma, liver cancer, or stomach cancer, and the inflammations include enteritis, myocarditis, obesity-related chronic inflammation of adipose tissue, liver, skeletal muscle, pancreatic islets, or brain tissue.

[0054] The abbreviations for the reaction reagents mentioned in the instructions are as follows:

[0055] DMF: N,N-dimethylformamide;

[0056] DIEA: N,N-diisopropylethylamine;

[0057] HCTU: 6-Chlorobenzotriazole-1,1,3,3-Tetramethylurea hexafluorophosphate;

[0058] DCM: Dichloromethane;

[0059] TFA: Trifluoroacetic acid;

[0060] HATU: 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate;

[0061] THF: Tetrahydrofuran;

[0062] NH2OH: Hydroxylamine;

[0063] EtOH: Ethanol;

[0064] HCl: hydrochloric acid;

[0065] TEA: Triethylamine;

[0066] CMFDA: 5-Chloromethylfluorescein diacetate.

[0067] Detailed description of the invention

[0068] Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0069] "Alkyl" refers to a saturated aliphatic hydrocarbon group. It includes straight-chain or branched groups with 1 to 20 carbon atoms. Preferably, it is a medium-sized alkyl group containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, etc. The alkyl group can be substituted or unsubstituted.

[0070] "Cx-y" is intended to include groups containing x to y carbons in the chain. For example, the term "C 1-4 "Alkyl" refers to a saturated hydrocarbon group, including straight-chain alkyl and branched-chain alkyl containing 1 to 4 carbons.

[0071] "Halogen" indicates fluorine, chlorine, bromine, or iodine.

[0072] The term "optionally" means that the event or situation described below may or may not occur, and the description includes both the possibility that the event or situation may or may not occur, and the description includes both the possibility that the event or situation may occur and the possibility that it does not occur.

[0073] In some implementations, "replaced by one or more groups" means that one, two, three, or four hydrogen atoms of a specified atom or group are replaced by the same or different groups selected from a specified range of groups.

[0074] The definitions and conventions of stereochemistry used in this invention are generally referenced in the following literature: SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984); McGraw-Hill Book Company, New York; and Elielj E. and Wilenj S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994.

[0075] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0076] "Pharmaceutically acceptable salts" refers to those salts that retain the bioavailability and properties of the parent compound.

[0077] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of this invention or their pharmaceutically acceptable salts, solvates, hydrates or prodrugs with other chemical components, such as pharmaceutically acceptable carriers.

[0078] "Solvate" refers to an association formed by one or more solvent molecules with the compounds of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecules are water.

[0079] "Pharmaceutical carrier" refers to the inactive component in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the given compound.

[0080] The beneficial effects of this invention are:

[0081] This invention discloses a compound having the structure of formula (I) or its stereoisomer or pharmaceutically acceptable salt. Using the fluorescent inhibitor CMFDA of GSTO1 as a specific probe, the relative occupancy rate of the compound with respect to GSTO1 was obtained by competing with the test compound. The compound exhibits high occupancy, high affinity for GSTO1, and strong inhibitory effect. The competitive GSTO1 protein pull-down method reveals the excellent ability and uses of this type of compound to inhibit GSTO1 activity, as well as its application in drug preparation. Attached Figure Description

[0082] Figure 1 The results of fluorescent gel assays for GSTO1 binding activity of the example compounds and comparative compounds in HCT116 cell lysates are shown.

[0083] Figure 2 The results of fluorescent gel assays for GSTO1 binding activity of the example compounds and comparative compounds in HeLa cell lysates are shown.

[0084] Figure 3 The results show the results of the example compounds and the comparative compounds competing for GSTO1 protein pull-down in A549 cells;

[0085] Figure 4The results show the results of the example compounds and the control compounds competing for GSTO1 protein pull-down in HeLa cells. Detailed Implementation

[0086] The present invention will be further illustrated below with reference to specific embodiments. These embodiments are implemented under the premise of the technical solution of the present invention. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0087] Example 1

[0088] Synthesis of Compound 1:

[0089]

[0090] In a reaction flask, 1a (500 mg, 2.45 mmol) and (2R, 4R)-1-tert-butoxycarbonyl-4-hydroxy-pyrrolidine-2-carboxylic acid (566.36 mg, 2.45 mmol) were added and dissolved in DMF (10 mL). Then, DIEA (633.08 mg, 4.90 mmol) and HCTU (1.52 g, 3.67 mmol) were added, and the mixture was stirred at 80 °C for 2 hours. LC-MS confirmed complete consumption of 1a. The temperature was then increased to 110 °C, and the reaction continued for 10 hours. LC-MS detected the appearance of the target molecular weight. The reaction was quenched with water (10 mL), and the mixture was extracted three times with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography was performed to separate the petroleum ether:ethyl acetate ratio (1:1). Evaporation to dryness yielded a yellow solid 1b (820 mg, 2.05 mmol), yield: 83.83%.

[0091] Dissolve 1b (400 mg, 1.00 mmol) in DCM (4 mL) in a reaction flask, then add TFA (0.4 mL) and stir at 20 °C for 12 hours. The reaction was confirmed to be complete by LC-MS. The pH was adjusted to greater than 7 by adding DIEA (3 mL), and the solution was evaporated to dryness to obtain a yellow oily crude product 1c (300 mg).

[0092] Dissolve 1c (150 mg, 501.26 μmol) in DCM (3 mL) in a reaction flask. Add DIEA (129.57 mg, 1.00 mmol) and acryloyl chloride (45.37 mg, 501.26 μmol) under ice bath conditions, stirring for 1 hour. LC-MS analysis confirmed the reaction was complete. The solution was evaporated to dryness and purified by liquid chromatography to give compound 1 (100.6 mg, 281.44 μmol) as a white solid, yield: 56.15%.

[0093] MS(ESI+): m / z 354.1 [M+H]+ ;

[0094] 1 H NMR (400MHz, DMSO-d6) δ=8.20 (d, J=8.0Hz, 2H), 7.92 (d, J=8.0Hz, 2H), 6.71-6.51 (m, 1H), 6.15 (dd, J=2.0, 16.8Hz, 1H), 5.73 (br d, J=9.6Hz, 1H), 5.38 (br s, 1H), 5.16 (br s, 1H), 4.53 (br s, 1H), 3.93 (br d, J=1.6Hz, 1H), 3.67 (dd, J=2.0, 10.8Hz, 1H), 2.40 (br s, 1H), 2.24 (br d, J=4.4Hz, 1H).

[0095] Example 2-13

[0096] Synthesis of compound 2-13:

[0097] The synthesis of the following compounds is based on the preparation of compound 1, and the relevant characterization is shown in Table 1.

[0098] Table 1

[0099]

[0100]

[0101] Example 14

[0102] Synthesis of compound 14:

[0103]

[0104] In a reaction flask, tert-butoxycarbonyl-D-proline (527.2 mg, 2.45 mmol) was dissolved in DCM (5 mL) and DMF (2.5 mL). Then, HATU (1.02 g, 2.69 mmol) and DIEA (949.6 mg, 7.35 mmol) were added, and the mixture was stirred at 20 °C for 10 minutes. Next, 14a (500 mg, 2.45 mmol) was added to the reaction mixture, and the mixture was stirred at 20 °C for 1.5 hours. The reaction was confirmed to be complete by LC-MS. The solution was evaporated to dryness, and extracted twice with water (30 mL) and ethyl acetate (20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography was used to separate the solid 14b (650 mg, 1.62 mmol), yield: 66.12%.

[0105] 1H NMR (400MHz, CDCl3) δ = 9.77-9.63 (m, 1H), 9.14-8.93 (m, 1H), 7.93 (br d, J=8.0Hz, 2H), 7.69 (brd, J=8.0Hz, 2H), 4.51-4.36 (m, 1H), 3.58-3.35 (m, 2H), 2.45-2.16 (m, 1H), 1.87 (br s, 3H), 1.51 (s, 9H)

[0106] 14b (17b) (300 mg, 747.42 μmol) was dissolved in THF (10 mL) in a reaction flask, and Lloyd's reagent (1.51 g, 3.74 mmol) was added. The mixture was stirred at 20 °C for 10 hours. The reaction was confirmed to be complete by LC-MS. The solution was evaporated to dryness and separated by column chromatography to give a yellow solid 14c (270 mg, 675.96 μmol), yield: 90.44%.

[0107] 14c (250 mg, 625.89 μmol) was dissolved in DCM (2.5 mL), and TFA (0.5 mL) was added. The mixture was stirred at 20 °C for 1 hour. LC-MS confirmed the reaction was complete. The solution was evaporated to dryness and dissolved in DCM (2.5 mL). Then, DIEA (106.3 mg, 822.6 μmol) and acryloyl chloride (74.45 mg, 822.57 μmol) were added. The mixture was stirred at 20 °C for 0.5 hours. LC-MS confirmed the reaction was complete. The solution was evaporated to dryness and purified by HPLC to give a yellow solid compound 14 (100.6 mg, 281.44 μmol), yield: 63.37%.

[0108] MS(ESI+): m / z 354.1 [M+H] + ;

[0109] 1 H NMR (400MHz, DMSO-d6) δ = 8.15 (d, J = 8.0Hz, 2H), 7.87 (d, J = 8.0Hz, 2H), 6.74-6.46 (m, 1H), 6.14 (brd, J = 2.0Hz, 1H), 5.89-5.73 (m, 1H), 5.67-5.46 (m, 1H), 3.89-3.79 (m, 1H), 3.73-3.54 (m, 1H), 2.39-2.20 (m, 2H), 2.14-1.98 (m, 2H).

[0110] Examples 15-16

[0111] Synthesis of compounds 15-16:

[0112] The synthesis of the following compounds is based on the preparation of compound 14, and the relevant characterization is shown in Table 2.

[0113] surface,

[0114]

[0115] Example 17

[0116] Synthesis of compound 17:

[0117]

[0118] 17a (1 g, 7.80 mmol) was dissolved in ethanol (10 mL) in a reaction flask, followed by the addition of hydroxylamine hydrochloride (542.34 mg, 7.80 mmol) and TEA (1.74 mL). The mixture was stirred at 80 °C for 3 hours. The reaction was confirmed to be complete by LC-MS. The solution was evaporated to dryness, and DCM (20 mL) and water (20 mL) were added. The mixture was filtered to obtain a solid, which was dried to give a yellow solid 17b (1 g, 6.20 mmol). Yield: 79.51%.

[0119] 1 H NMR (400MHz, DMSO-d6) δ = 10.02 (s, 1H), 7.92-7.77 (m, 4H), 6.00 (s, 2H)

[0120] In a reaction flask, 17b (0.9 g, 5.58 mmol) was dissolved in DMF (10 mL), and HCTU (3.47 g, 8.38 mmol), (tert-butoxycarbonyl)-D-proline (1.44 g, 6.70 mmol), and DIEA (1.44 g, 11.17 mmol) were added. The mixture was stirred at 80 °C for 2 hours. LC-MS detected complete consumption of 6b, and the temperature was increased to 110 °C for another 6 hours. LC-MS detected the appearance of the target molecular weight. The reaction mixture was extracted three times with water (50 mL) and ethyl acetate (50 mL), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The mixture was then separated by column chromatography (petroleum ether:ethyl acetate = 1:1) and evaporated to dryness to give a yellow solid 17c (1 g, 2.94 mmol), yield: 80.99%.

[0121] 1 H NMR (400MHz, DMSO-d6) δ = 8.21-8.13 (m, 2H), 8.05 (d, J = 8.4Hz, 2H), 5.21-5.06 (m, 1H), 3.58-3.49 (m, 1H), 3.43 ( td, J=7.2, 10.4Hz, 1H), 2.45-2.31(m, 1H), 2.24-2.02(m, 2H), 1.88-1.74(m, 1H), 1.39(s, 3H), 1.26-1.11(m, 6H)

[0122] 17c (200 mg, 587.59 μmol) was dissolved in DCM (4 mL), and TFA (0.6 mL) was added. The mixture was stirred at 20 °C for 1 hour. LC-MS confirmed the reaction was complete. The solution was evaporated to dryness, and the reaction was quenched by adding DIEA (2.5 mL) under ice bath conditions. The solution was then evaporated to dryness to obtain 17d. 17d was dissolved in DCM (2 mL), followed by the addition of DIEA (129.10 mg, 998.92 μmol) and acryloyl chloride (45.21 mg, 499.46 μmol). The mixture was stirred at 20 °C for 0.5 hours. LC-MS confirmed the reaction was complete. The solution was evaporated to dryness, and the solution was purified by liquid chromatography to give a yellow solid compound 17 (34.8 mg, 110.13 μmol), yield: 22.05%.

[0123] MS(ESI+): m / z 295.1 [M+H] + ;

[0124] 1 H NMR (400MHz, DMSO-d6) δ = 8.19-8.12 (m, 2H), 8.08-8.01 (m, 2H), 6.74-6.49 (m, 1H), 6.15 (dd, J = 2.0, 16.8Hz, 1H), 5.82- 5.73 (m, 1H), 5.35 (dd, J=3.6, 8.8Hz, 1H), 3.92-3.83 (m, 1H), 3.78-3.50 (m, 1H), 2.47-2.32 (m, 1H), 2.19-1.86 (m, 3H).

[0125] Examples 18-25

[0126] Synthesis of compounds 18-25

[0127] The synthesis of the following compounds is based on the preparation of compound 17, and the relevant characterization is shown in Table 3.

[0128] Table 3

[0129]

[0130] Comparative Example 1

[0131] Synthesis of compound 26:

[0132]

[0133] 26a (500 mg, 2.45 mmol) and (2R, 4S)-1-tert-butoxycarbonyl-4-hydroxy were added to the reaction flask.

[0134] 2-pyrrolidine-2-carboxylic acid (566.36 mg, 2.45 mmol) was dissolved in DMF (10 mL). Then, DIEA (633.08 mg, 4.90 mmol) and HCTU (1.52 g, 3.67 mmol) were added, and the mixture was stirred at 80 °C for 2 hours. LC-MS analysis showed complete consumption of 26a. The temperature was then increased to 110 °C, and the reaction continued for 10 hours. LC-MS analysis showed the appearance of the target molecular weight. The reaction was quenched with water (10 mL), and the mixture was extracted three times with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Column chromatography was performed to separate the petroleum ether:ethyl acetate ratio (1:1). Evaporation to dryness yielded a yellow solid, 26b (820 mg, 2.05 mmol), with a yield of 83.83%.

[0135] Dissolve 26b (400 mg, 1.00 mmol) in DCM (4 mL) in a reaction flask, then add TFA (0.4 mL) and stir at 20 °C for 12 hours. The reaction was confirmed to be complete by LC-MS. Adjust the pH to greater than 7 by adding DIEA (3 mL) and evaporate to dryness to obtain a yellow oily crude product 26c (300 mg).

[0136] Dissolve 26c (150 mg, 501.26 μmol) in DCM (3 mL) in a reaction flask. Add DIEA (129.57 mg, 1.00 mmol) and acryloyl chloride (45.37 mg, 501.26 μmol) under ice bath conditions, stirring for 1 hour. LC-MS analysis confirmed the reaction was complete. The solution was evaporated to dryness and purified by liquid chromatography to give a white solid, compound 26 (100.6 mg, 281.44 μmol), yield: 56.15%.

[0137] LCMS(ESI+): m / z 354.1[M+H]+

[0138] 1 H NMR (400MHz, DMSO-d6) δ = 8.20 (d, J = 8.0Hz, 2H), 7.94 (d, J = 8.4Hz, 2H), 6.72- 6.46 (m, 1H), 6.15 (dd, J=2.0, 16.8Hz, 1H), 5.84-5.61 (m, 1H), 5.39 (d, J=3.2 Hz, 1H), 5.33 (t, J=8.0Hz, 1H), 4.58-4.39 (m, 1H), 3.94 (dd, J=4.4, 10.8Hz, 1 H), 3.72-3.58 (m, 1H), 2.47-2.32 (m, 1H), 2.18 (ddd, J=4.8, 8.4, 13.2Hz, 1H)

[0139] Comparative Examples 2-4

[0140] Comparative Example 2, Compound 27, Comparative Example 3, Compound 28, Comparative Example 4, and Compound 29 were synthesized using the same method as that used to synthesize Compound 26 in Comparative Example 1. The relevant characterizations are shown in Table 4.

[0141] Table 4

[0142]

[0143] Comparative Example 5

[0144] Comparative Example 5, compound 30, was synthesized according to the synthesis methods of Examples 14-16, and the relevant characterization is shown in Table 5.

[0145] Table 5

[0146]

[0147] Comparative Example 6

[0148] Comparative Example 6, compound 31, was synthesized according to the synthesis methods of Examples 17-25, and the relevant characterization is shown in Table 6.

[0149] Table 6

[0150]

[0151] Comparative Example 7

[0152] Synthesis of compound 32:

[0153]

[0154] In a reaction flask, 32a (1 g, 4.90 mmol) and (2R, 4R)-1-tert-butoxycarbonyl-4-hydroxy-pyrrolidine-2-carboxylic acid (1.13 g, 4.90 mmol) were added and dissolved in DMF (30 mL). Then, DIEA (1.27 g, 9.80 mmol) and HCTU (3.04 g, 7.35 mmol) were added, and the mixture was stirred at 80 °C for 2 hours. The reaction was confirmed to be complete by LC-MS, and the solution was evaporated to dryness to obtain crude product 32b. 32b was completely dissolved in DMF (30 mL), and DIEA (928.96 mg, 7.19 mmol) was added. The mixture was stirred at 110 °C for 2 hours. The reaction was confirmed to be complete by LC-MS. The reaction was quenched by adding water (10 mL), diluted with ethyl acetate (50 mL), and extracted three times with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The petroleum ether:ethyl acetate ratio of 1:1 was separated by column chromatography and evaporated to dryness to give a yellow oily substance 32c (1.4 g, 23.51 mmol), yield: 73.16%.

[0155] 32c (600 mg, 1.50 mmol) was dissolved in DCM (30 mL) in a reaction flask. TEA (0.4 mL) and MsCl (670 mg, 5.85 mmol) were added under ice bath conditions, and the mixture was stirred at 20 °C for 1 hour. The reaction was confirmed to be complete by LC-MS. The reaction was quenched with water (20 mL), and the mixture was extracted twice with DCM (20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Petroleum ether:ethyl acetate (1:1) was separated by column chromatography, and the solid was evaporated to dryness to give 32d (450 mg, 942.50 mmol), yield: 62.73%.

[0156] t-BuOK (126.91 mg, 1.13 mmol) was dissolved in NMP (3 mL) in a reaction flask, and dimethyl malonate (149.42 mg, 1.13 mmol) was added. The mixture was stirred at 40 °C for 0.5 h. 32d (180 mg, 377.00 μmol) was added and stirred at 80 °C for 1 h. The reaction was confirmed to be complete by LC-MS. The mixture was extracted three times with water (5 mL) and ethyl acetate (3 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Preparative thin-layer chromatography (TLC) with petroleum ether:ethyl acetate (1:1) yielded a pale yellow oily product 32e (70 mg, 136.33 μmol), yield: 36.16%.

[0157] In a reaction flask, 32e (70 mg, 136.33 μmol) was dissolved in DMSO (1 mL) and water (0.04 mL), and NaCl (15.93 mg, 272.66 μmol) was added. The mixture was stirred at 130 °C for 12 hours. The reaction was confirmed to be complete by LC-MS. The reaction was quenched by adding water (10 mL), diluted with ethyl acetate (10 mL), and extracted three times with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow oily crude product 32f (80 mg).

[0158] 32f (70 mg, 153.70 μmol) was dissolved in methanol (2 mL) in a sealed tube, and NH3 / MeOH (700 μL) was added. The mixture was stirred at 60 °C for 12 hours. The reaction was confirmed to be complete by LC-MS, and the product was evaporated to dryness to give 32 g (70 mg) of a yellow oily crude product.

[0159] 32 g (60 mg, 136.24 μmol) was dissolved in DCM (2 mL) in a reaction flask, and TFA (180 μL) was added. The mixture was stirred at 20 °C for 1 hour. LC-MS confirmed the reaction was complete. TEA (2 mL) was added to adjust the pH to greater than 7, and the mixture was evaporated to dryness. DCM (2 mL) was then added, followed by DIEA (62.67 mg, 484.87 μmol) and acryloyl chloride (14.63 mg, 161.62 μmol). The mixture was stirred at 20 °C for 0.5 hours. LC-MS confirmed the reaction was complete. Water (5 mL) was added to quench the reaction. The mixture was extracted three times with DCM (2 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness. The mixture was purified by liquid chromatography to give a white solid compound 32 (13.5 mg, 33.62 μmol), yield: 20.80%.

[0160] MS(ESI+): m / z 395.1 [M+H]+

[0161] 1 H NMR (400MHz, DMSO-d6) δ = 8.20 (d, J = 8.4Hz, 2H), 7.92 (d, J = 8.4Hz, 2H), 7.32-6.86 (m, 1H), 6.81-6.48 (m, 2H), 6.25-6.02 (m, 1H), 5.73 (br d, J=8.4Hz, 1H), 5.52-5.02 (m, 1H), 4.14-3.35 (m, 2H), 2.94-2.62 (m, 1H), 2.47-2.29 (m, 2H), 2.29 (br d, J=1.6Hz, 1H), 2.11-1.78 (m, 1H)

[0162] Example 1: Assessment of GSTO1 binding activity in cell lysate

[0163] Table 7 Preparation of cell lines, culture media and test compounds

[0164] cell lines culture medium HCT116 McCoy's 5a+10% FBS Hela DMEM + 10% FBS

[0165] As shown in Table 7, the cell lines to be tested were placed in a culture medium containing 10% heat-inactivated FBS (fetal bovine serum), 2 mM L-glutamine, 100 U / mL penicillin, and 100 g / mL streptomycin (components are shown in the table) and cultured in a cell culture incubator at 37°C and 5% CO2. The medium was changed every two days. After the cells reached 80% confluence, they were digested with trypsin, passaged, and kept in a good logarithmic growth phase.

[0166] Detection method:

[0167] HCT116 cells in logarithmic growth phase at 5*10 6Cells were seeded per 6cm cell culture dish and cultured for 24 hours. Cells were washed with PBS (phosphate-buffered saline) and serum-free medium, respectively. Serum-free medium containing the test sample (concentrations shown in the table) was prepared and added to the culture dishes. Cells were then incubated for another 2 hours. Next, 0.5μM CMFDA was added, and the cells were incubated for another 1 hour. The cell culture dishes were then placed on ice, and the cells were washed twice with cold PBS. PBS was then added again to scrape off the cells and collect them.

[0168] The obtained cells were added to cell lysis buffer [PBS (Thermo Fisher Scientific), 1% IGEPA-CA-630 (Sigma-Aldrich), 0.2% SDS (Sigma-Aldrich), 1% EDTA-free protease inhibitor mixture (Sigma-Aldrich), 0.1% Benzonase (Beyotime)], and after sonication, were centrifuged at high speed (20000g, 30 minutes) in a benchtop centrifuge at 4°C. The supernatant was collected and the concentration was adjusted to 2 mg / mL with cell lysis buffer. 500 μL of protein solution was added to a final concentration of 200 M fluorescein azide, 2.5 mM sodium ascorbate (Sigma-Aldrich), 25 mM BTTAA (CONFLUORE), and 12.5 mM CuSO4 (Innochem) at room temperature, and reacted for 1 hour. After the above reaction, the samples were separated by SDS-PAGE and electrophoresed at 150 V for approximately 1 hour. The gel fluorescent bands were visualized using ChemiDoc imaging scanning, followed by staining with Coomassie blue, as shown. Figure 1 and Figure 2 As shown, reference compound C1-27: (CAS:568544-03-6), Lit.44: (CAS: 2290641-18-6).

[0169] Relative occupancy rate (%) = 100 - (target band fluorescence signal in sample group / target band fluorescence signal in blank group) * (average Cochlear staining signal in sample group / average Cochlear staining signal in blank group) * 100

[0170] Table 8. Relative occupancy of compounds to GSTO1

[0171]

[0172]

[0173] The results are shown in Table 8. Using CMFDA as a specific probe, the fluorescence intensity on the SDS-PAGE gel was analyzed based on the principle of competition between the test compound and GSTO1. The relative occupancy rate of the compound on GSTO1 was obtained. The higher the occupancy rate, the higher the affinity of the compound with GSTO1 and the stronger the inhibitory effect.

[0174] Comparative Experiment Example 1: Evaluation of GSTO1 binding activity in cell lysate of the comparative compound

[0175] The reagents and detection methods are as in Experiment 1, and the relevant characterizations are shown in Table 9.

[0176] Table 9. Relative occupancy of comparative compounds for GSTO1

[0177]

[0178] The results are shown in Table 9. Using CMFDA as a specific probe, the fluorescence intensity on the SDS-PAGE gel was analyzed based on the principle of competition between the comparative compounds and GSTO1 to obtain the relative occupancy rate of the compounds. Among them, the relative occupancy rates of comparative compounds 26, 27, 29, 30, and 31 were 0, while the relative occupancy rates of comparative compounds 28 and 32 were weak, indicating low affinity for GSTO1 and weak inhibitory effect.

[0179] Experiment 2: Assessment of GSTO1 binding activity in live cells, detection of GSTO1 protein pull-down inhibition.

[0180] Table 10 Cell Culture and Preparation of Test Compounds

[0181] cell lines culture medium Hela DMEM + 10% FBS A549 F-12K+10% FBS

[0182] As shown in Table 10, the cell lines to be tested were placed in a culture medium containing 10% heat-inactivated FBS (fetal bovine serum), 2 mM L-glutamine, 100 U / mL penicillin, and 100 g / mL streptomycin (components are shown in the table) and cultured in a cell culture incubator at 37°C and 5% CO2. The medium was changed every two days. After the cells reached 80% confluence, they were trypsinized, passaged, and kept in a good logarithmic growth phase.

[0183] All samples were dissolved in DMSO.

[0184] Detection method:

[0185] A549 cells in logarithmic growth phase grow at a rate of 1*10 7 Cells were seeded per 10cm cell culture dish and cultured for 24 hours. Cells were washed with PBS and serum-free medium, respectively. GST01 protein enrichment reagents were prepared. Add serum-free culture medium containing the sample to be tested (concentrations shown in the table) to a culture dish, and incubate the cells in an incubator for 1 hour. Then place the cell culture dish on ice and wash the cells twice with cold PBS. Add PBS again, scrape off the cells, and collect them.

[0186] The obtained cells were added to cell lysis buffer [PBS (ThermoFisher Scientific), 1% IGEPEAL-CA-630 (Sigma-Aldrich), 0.2% SDS (Sigma-Aldrich), 1% EDTA-free protease inhibitor mixture (Sigma-Aldrich), 0.1% Benzonase (Beyotime)], and after sonication, were centrifuged at high speed (20000g, 30 minutes) in a benchtop centrifuge at 4°C. The supernatant was collected and the concentration was adjusted to 2 mg / mL with cell lysis buffer. 500 μL of protein solution was added to a final concentration of 200 M biotin azide, 2.5 mM sodium ascorbate (Sigma-Aldrich), 25 mM BTTAA (CONFLUORE), and 12.5 mM CuSO4 (Innochem) at room temperature, and reacted for 1 hour. After the above reaction is complete, transfer the protein solution to a 15 mL centrifuge tube. Add 4 volumes of methanol, vortex to mix thoroughly, 1 volume of chloroform, vortex to mix thoroughly, and 3 volumes of water, vortex to mix thoroughly. Centrifuge at 3000 rpm for 10 minutes at 4°C, retaining the protein precipitate (round discs). Add 1 mL of pre-cooled methanol, vortex or sonicate (for about 5 seconds) to break up the precipitate, transfer to a 1.5 mL centrifuge tube, centrifuge at 3000 rpm for 3 minutes at 4°C, and discard the supernatant. Repeat the above steps once. Air-dry the protein precipitate for 2 minutes to completely remove methanol.

[0187] The protein precipitate was resuspended in 3 mL of 0.05% SDS / PBS, and 200 μL of streptavidin-conjugated magnetic beads (ThermoFisher Scientific) was added. The mixture was incubated at room temperature for 3 hours by rotation. After enrichment, the protein was separated by SDS-PAGE and transferred to an NC membrane. After transfer, the NC membrane was immersed in Ponceau S staining solution, and the Ponceau S signal was read. The NC membrane was then incubated in 10 mL of 5% BSA / TBST at room temperature for 1 hour. The membrane was then incubated overnight at 4°C with the target protein antibody GSTO1 polyclonal antibody (Proteintech). Finally, the membrane was incubated with the secondary antibody (Alexa Fluor 488 labeled Goat Anti-Rabbit IgG, Beyotime) for 1 hour. The fluorescence signal of the secondary antibody was scanned using ChemiDoc imaging. Figure 3 , Figure 4 As shown. The relative occupancy rate of the sample to be tested is calculated using the following formula:

[0188] Inhibition rate (%) = 100 - (Target band signal in sample group / Target band signal in blank group) * 100

[0189] Table 11 Inhibition rate of compounds on GSTO1 downregulation

[0190]

[0191]

[0192] As shown in Table 11, the affinity of the compounds of the examples for GSTO1 in living cells was revealed by a competitive protein pull-down method. The inhibition rates of compounds C1-27, Lit.44, and the compounds of the examples on GSTO1 pull-down in two cell lines (A549 & Hela) were tested, and the results showed that the compounds of the examples had excellent ability to inhibit GSTO1 activity.

[0193] Comparative Experiment Example 2: Assessment of GSTO1 binding activity of the comparative compound in live cells, and detection of GSTO1 protein pull-down inhibition.

[0194] The reagents and detection methods are as in Experiment Example 2, and the relevant characterization is shown in Table 12.

[0195] Table 12 shows the inhibition rate of comparative compounds on GSTO1 downregulation.

[0196] Group compound Final concentration (μM) Inhibition rate (%) in A549 cells Inhibition rate (%) in HeLa cells Comparative Example 1 26 30 0 0 Comparative Example 2 27 30 0 0 Comparative Example 3 28 30 20 22 Comparative Example 4 29 30 0 0 Comparative Example 5 30 30 0 0 Comparative Example 6 31 30 0 0 Comparative Example 7 32 30 9 0

[0197] As shown in Table 12, the affinity of the comparative compounds for GSTO1 in live cells was revealed by a competitive protein pull-down method. The inhibition rates of the comparative compounds on GSTO1 pull-down in two cell lines (A549 & HeLa) were tested. The results showed that comparative compounds 26, 27, 29, 30, and 31 had an inhibition rate of 0, while comparative compounds 29 and 32 showed weak inhibitory activity against GSTO1. Example 3: Cell proliferation inhibition experiment using compounds, CCK-8 assay.

[0198] Table 13 Preparation of Cell Lines, Culture Media and Compounds

[0199] cell lines culture medium HEK293T DMEM + 10% FBS A549 F-12K+10% FBS HCT116 McCoy's 5a+10% FBS

[0200] As shown in Table 13, the cell lines to be tested were placed in a culture medium containing 10% heat-inactivated FBS (fetal bovine serum), 2 mM L-glutamine, 100 U / mL penicillin, and 100 g / mL streptomycin (components are shown in the table) and cultured in a cell culture incubator at 37°C and 5% CO2. The medium was changed every two days. After the cells reached 80% confluence, they were trypsinized, passaged, and kept in a good logarithmic growth phase.

[0201] IC 50 test:

[0202] Collect cells in the logarithmic growth phase, count them, resuspend the cells in complete culture medium, and adjust the cell concentration to 1*10⁻⁶. 4 Cells / mL were seeded into 96-well plates, with 100 μL of cell suspension added to each well. Cells were incubated at 37°C and 5% CO2 for 24 h.

[0203] Dilute the test compound to the set target concentration using culture medium, and add 25 μL / well to a 96-well plate. Incubate at 37°C and 5% CO2 for 48 h. Aspirate the culture medium, add complete culture medium containing 10% CCK-8, and incubate at 37°C and 5% CO2 for 4 h. Gently shake and measure the absorbance at 450 nm using a microplate reader. Calculate the cell growth inhibition rate of the compound using the following formula:

[0204] Cell growth inhibition rate (%) = [(Negative control - Sample) / (Negative control - Positive control)] * 100

[0205] IC was performed using the software Graphpad Prism 8 and the formulas log(inhibitor) vs. Normalized response-variable slope or log(inhibitor) vs. Response variable slope. 50 Curve fitting and IC calculation 50 The numerical values ​​and results are shown in Table 14.

[0206] Table 14

[0207] Group <![CDATA[IC 50 / μM]]> HEK293T A549 HCT116 Reference compound C1-27 2.33 5.69 9.81 Example 3 3 32.35 28.82 33.64 Example 6 6 46.08 40.74 45.96 Example 14 14 28.97 31.75 29.11 Example 17 17 37.62 42.68 47.92 Example 18 18 24.38 29.17 26.58 Example 22 22 36.32 42.52 44.52 Example 23 23 41.72 46.25 47.31 Example 25 25 47.05 42.65 46.53

[0208] The inhibitory effects of compound C1-27 and representative compounds on cell proliferation in three cell lines (MDA-MB-231, A549, and HEK293T) were tested and compared. The results showed that C1-27 was highly toxic, while the compound in the example had low cytotoxicity and better safety than C1-27.

[0209] The above specific embodiments further illustrate the purpose, technical solution and beneficial effects of this application. It should be understood that the above are only specific embodiments of this application and are not intended to limit the scope of protection of this application. Any modifications, equivalent substitutions, improvements, etc., made on the basis of the technical solution of this application should be included within the scope of protection of this application.

Claims

1. A compound of Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: in, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -C(R1)-, -CH2-, -CH-, -NR4-, -O-, or -S-, while L3 is selected from -C(R3)-, -CH(R3)-, or -CH2-. R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C groups. 1-4 Alkyl, wherein the C 1-4 The alkyl group is substituted by one or more hydrogens or halogens, wherein the chiral carbon attached to the -OH group is in the R configuration; R1 and R2 can form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 can form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is substituted by one or more hydrogens or halogens. R4 is selected from hydrogen, -OH, or C. 1-4 alkyl; n = 1 or 2; L4, L5, L6, L7, and L8 are each independently selected from -C-, -C(R5)-, -N-, -O-, and -S-; R5 is selected from hydrogen, halogen, -OH, cyano, carboxyl, or C. 14 alkyl; R6 is selected from hydrogen, halogens, and C. 1-4 Alkyl or hydroxyl; R8 is selected from halogen, cyano, and C. 1-4 Alkoxy or C 1-4 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more hydrogens, halogens, or C atoms. 1-4 Alkyl substitution; R9 is selected from hydrogen, halogens, and C. 14 Alkyl or dimethylaminosulfonyl; R7, R 10 Selected from hydrogen, halogen or C 1-4 alkyl; R 11 It is selected from hydrogen, halogen or cyano.

2. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by formula I-1: Among them, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -C(R1)-, -CH2-, -CH-, -NR4-, -O-, or -S-. L3 is selected from -C(R3)-, -CH(R3)-, and -CH2-; R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C groups. 1-4 Alkyl, wherein the C 1-4 The alkyl group is substituted by one or more hydrogens or halogens, wherein the chiral carbon attached to the -OH group is in the R configuration; R1 and R2 can form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 can form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is substituted by one or more hydrogens or halogens. R4 is selected from hydrogen, -OH, or C. 1-4 alkyl; n = 1 or 2; L5, L7, and L8 are each independently selected from -C(R5)-, -N-, -O-, and -S-, respectively, with R5 selected from hydrogen. R6 is selected from hydrogen, halogens, and C. 1-4 Alkyl or hydroxyl; R8 is selected from halogen, cyano, and C. 1-4 Alkoxy or C 1-4 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more hydrogens, halogens, or C atoms. 1-4 Alkyl substitution; R9 is selected from hydrogen, halogens, and C. 14 Alkyl or dimethylaminosulfonyl; R 11 It is selected from hydrogen, halogen or cyano.

3. The compound according to claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, Selected from:

4. The compound according to claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by formula I-2: When n=1, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -CH2-, -CH- or -S-, and L3 is selected from -CH(R3)- or -CH2-. R1, R2, and R3 are independently selected from hydrogen, halogens, or -OH, wherein the chiral carbon attached to -OH is in the R configuration, or R1 and R2 form an arbitrarily substituted 6-membered aromatic ring, or R2 and R3 form an arbitrarily substituted 6-membered aromatic ring, wherein the 6-membered aromatic ring is optionally substituted by one or more hydrogens or halogens.

5. The compound according to claim 4, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The 6-membered aromatic ring is a benzene ring.

6. The compound according to claim 2 or 4, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, Selected from:

7. The compound according to claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, When n=2, L1 and L2 are each independently selected from -C(R1)2-, -CH(R2)-, -CH2-, -NR4-, -O- or -S-, and L3 is selected from -CH(R3)- or -CH2-. R1, R2, and R3 are independently selected from hydrogen, halogens, -OH groups, or any substituted C atoms. 1-4 Alkyl group, wherein the chiral carbon attached to the -OH group has an R configuration, wherein the C 1-4 Alkyl groups may optionally be substituted with one or more hydrogen atoms or halogens.

8. The compound according to claim 2 or 7, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, Selected from:

9. The compound according to claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R8 is selected from halogen, cyano, and C. 1-4 Alkoxy or C 14 Alkyl, wherein the C 1-4 Alkoxy, C 1-4 Alkyl groups are optionally surrounded by one or more of the following hydrogens, halogens, or C atoms. 1-4 Alkyl substitution; R6 is selected from hydrogen or hydroxyl; R9 is selected from hydrogen, halogen or dimethylaminosulfonyl.

10. The compound according to claim 2 or 9, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, Selected from:

11. The compound according to any one of claims 1-10, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is selected from any of the following compounds:

12. A pharmaceutical composition, characterized in that, It includes the compound, stereoisomer or pharmaceutically acceptable salt thereof of any one of claims 1-11, and one or more pharmaceutically acceptable carriers, diluents or excipients.

13. Use of any compound of claims 1-12, or a solvate, metabolite, cocrystal, prodrug, or combination thereof, in a medicament for the treatment or prophylactic treatment of a disease regulated by a GSTO1 inhibitor.

14. The use according to claim 13, wherein the disease includes cancer, inflammation, or age-related macular degeneration.

15. The disease according to claim 14, wherein the cancer includes breast cancer, ovarian cancer, cervical cancer, pancreatic cancer, colorectal cancer, esophageal squamous cell carcinoma, liver cancer, or gastric cancer, and the inflammation includes enteritis, myocarditis, obesity-related chronic inflammation of adipose tissue, liver, skeletal muscle, pancreatic islets, or brain tissue.

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