SELENIUM-CONTAINING ISOXAZOLAMINE COMPOUND, PREPARATION METHOD THEREFOR, AND USE THEREOF

Abstract

The present invention disclosed a series of novel selenium-containing isoxazolamine derivatives as shown in formula I, which could regulate the generation and/or activity of TNF-α and ferroptosis-like cell death. The present invention also disclosed the preparation method and the use thereof in preparing a drug for treating the diseases mediated by TNF-α and/or iron-dependent cell death.

Claims

1. A selenium-containing isoxazolamine derivative having a structure of general formula (I), a pharmaceutically acceptable salt, a solvate, a polymorph, a stereoisomer, an isotopic compound, or a metabolite thereof; ##STR00022## in the general formula (I), each of R.sub.1, R.sub.2, R.sub.3 and R.sub.4 is independently selected from H, D, halogen, hydroxyl, amino, nitro, cyano, carboxyl, seleno, mercapto, (C.sub.1-C.sub.8) alkylselenyl, (C.sub.1-C.sub.8) alkylselenyl (C.sub.1-C.sub.8) alkylamino, (C.sub.2-C.sub.8) alkenylselenyl, α-(C.sub.1-C.sub.8) alkylselenyl amino acid, α-(C.sub.1-C.sub.8) alkylselenyl formyl amino acid, (C.sub.0-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkylselenyl, (C.sub.0-C.sub.8) alkylaminoformylselenyl, (C.sub.0-C.sub.8) alkylaminoformyl, arylselenyl, (C.sub.0-C.sub.8) alkoxyl (C.sub.1-C.sub.8) alkylselenyl, (C.sub.0-C.sub.8) alkoxyformyl (C.sub.1-C.sub.8) alkylselenyl, (C.sub.0-C.sub.8) alkoxyformyl C.sub.1-C.sub.8 alkoxyl, halo (C.sub.1-C.sub.8) alkylselenyl, C.sub.1-C.sub.8 alkanesulfonyl, (C.sub.1-C.sub.8) alkanesulfonamido, (C.sub.0-C.sub.8) alkylaminosulfonyl, (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkoxyl, (C.sub.0-C.sub.8) alkylethynyl, (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkylacyloxy, (C.sub.1-C.sub.8) alkoxyl (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkoxyl (C.sub.1-C.sub.8) alkyl, (C.sub.1-C.sub.8) alkylamino, (C.sub.0-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkyl, aryl, aryl (C.sub.1-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkyl, amidino, guanidino, arylsulfonamido, arylaminosulfonyl, benzoyl, (C.sub.0-C.sub.8) alkylselenyl formyl, aryl (C.sub.1-C.sub.8) alkylamino, aryl (C.sub.1-C.sub.8) alkylamido, (C.sub.1-C.sub.8) alkoxyformyl, (C.sub.1-C.sub.8) alkylamido, (C.sub.1-C.sub.8) alkylamino, (C.sub.0-C.sub.8) alkylselenyl formamido, arylselenyl (C.sub.1-C.sub.8)alkylamido, selenylcyano (C.sub.1-C.sub.8) alkylamido, benzoisoselenidazolone amino, benzoisoselenidazolone amino (C.sub.1-C.sub.8) alkylamido, benzoisoselenidazolone amino (C.sub.1-C.sub.8) alkanesulfonamido, (C.sub.0-C.sub.8) alkylamino selenyl, (C.sub.0-C.sub.8) alkylaminoformyl, (C.sub.0-C.sub.8) alkylamino formylselenyl, (C.sub.1-C.sub.8) alkylaminoformyloxyl, (C.sub.1-C.sub.8) alkylaminoformyl, (C.sub.1-C.sub.8) alkylaminoformyloxy, arylaminoformamido, arylaminoformyl, arylaminoformyloxy, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrazinyl, quinolinyl, pyrimidinyl, pyrimidinylamino, thiazolyl, thienyl, furanyl, pyrrolyl or absent; wherein, the aryl groups of R.sub.1, R.sub.2, R.sub.3 and R.sub.4 described are phenyl or are phenyl which independently substituted with 1-4 halogen, hydroxy, nitro, cyano, amino, trifluoromethyl, carboxyl, (C.sub.0-C.sub.8) alkylaminosulfonyl, (C.sub.1-C.sub.8) alkanesulfonamido, (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkoxyl groups; wherein, the benzoisoselenidazolone amino described is ##STR00023## Z is: ##STR00024## wherein Z is ##STR00025## R.sub.5 is selected from H, D, (C.sub.1-C.sub.8) alkylselenyl (C.sub.1-C.sub.8) alkyl, (C.sub.2-C.sub.8) alkenylselenyl (C.sub.1-C.sub.8) alkyl, selenocyanate (C.sub.1-C.sub.8) alkyl, ##STR00026## wherein Z is s ##STR00027## R.sub.5 is selected from H, halogen, hydroxyl, nitro, cyano, amino, trifluoromethyl, carboxyl, (C.sub.1-C.sub.8) alkanesulfonyl, amino sulfonyl, (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkoxyl; W is C or Se; wherein, when W is C, there is one selenium-containing substituent exists in the R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 group at least; and when W is Se, R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 could be any substitutes as described above; X is, O or not exist; Where bonds represented by is chemical bond or not exist.

2. The selenium-containing isoxazolamine derivatives having a structure of general formula (I), the pharmaceutically acceptable salt, the solvate, the polymorph, the stereoisomer, the isotopic compound, or the metabolite thereof according to claim 1, wherein the compounds having structures of general formula (I-a), (I-b), (I-c), (I-d) and/or (I-e): ##STR00028## in the general formula (I-a˜I-e), each of R.sub.1, R.sub.2, R.sub.3 and R.sub.4 is independently selected from H, D, halogen, hydroxyl, amino, nitro, cyano, carboxyl, (C.sub.0-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkylselenyl, (C.sub.0-C.sub.8) alkylaminoformyl (C.sub.1-C.sub.8) alkoxyl, amidino, guanidino, C.sub.1-C.sub.8 alkanesulfonyl, (C.sub.1-C.sub.8) alkanesulfonamido, (C.sub.0-C.sub.8) alkylaminosulfonyl, (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkyl, halo (C.sub.1-C.sub.8) alkoxyl, (C.sub.0-C.sub.8) alkylethynyl, (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkylacyloxy, (C.sub.1-C.sub.8) alkoxyl (C.sub.1-C.sub.8) alkoxyl, (C.sub.1-C.sub.8) alkoxyl (C.sub.1-C.sub.8) alkyl, (C.sub.1-C.sub.8) alkylamino, (C.sub.0-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkyl, aryl, aryl (C.sub.1-C.sub.8) alkylamino (C.sub.1-C.sub.8) alkyl, arylsulfonamido, arylaminosulfonyl, benzoyl, arylmethylamino, aryl formamido, (C.sub.0-C.sub.8) alkoxyformyl, (C.sub.1-C.sub.8) alkylamido, (C.sub.1-C.sub.8) alkylamino, (C.sub.0-C.sub.8) alkylaminoformamido, (C.sub.0-C.sub.8) alkylaminoformyl, arylaminoformamido, arylaminoformyloxy or absent; wherein, the aryl groups of R.sub.1, R.sub.2, R.sub.3 and R.sub.4 described are phenyl or are phenyl which independently substituted with 1-4 halogen, hydroxy, nitro, cyano, trifluoromethyl, carboxyl, aminosulfonyl, (C.sub.1-C.sub.6) alkyl, (C.sub.1-C.sub.6) alkoxyl groups; X is, O or not exist; where bonds represented by is chemical bond or not exist.

3. The selenium-containing isoxazolamine derivatives having a structure of general formula (I), the pharmaceutically acceptable salt, the solvate, the polymorph, the stereoisomer, the isotopic compound, or the metabolite thereof according to claim 1, wherein the compounds having a structure of general formula (I) is selected from the group consisting of ##STR00029## ##STR00030## ##STR00031## ##STR00032## ##STR00033##

4. A pharmaceutical composition, which comprises at least one substance selected from the group consisting of the selenium-containing isoxazolamine derivative of claim 1, the pharmaceutically acceptable salt, the solvate, the polymorph, the stereoisomer, the isotopic compound, or the metabolite thereof, as well as one or more pharmaceutically acceptable carriers, diluents or excipients.

5. A process for preparing the selenium-containing isoxazolamine derivatives having a structure of general formula (I) according to claim 1, comprising: (1) the first synthesis route for preparing some of the selenium-containing isoxazolamines represented by general formula I-a, I-b, I-c, I-d, and I-e ##STR00034## (2) the second synthesis route for preparing some of the selenium-containing isoxazolamines represented by general formula I-a, I-b, I-c, I-d, and I-e ##STR00035## (3) the third synthesis route for preparing tetravalent selenium compounds represented by general formula I-a, I-b, I-c, I-d, and I-e ##STR00036## in the first synthesis route for preparing the selenium-containing isoxazolamines represented by general formula I-a, I-b, I-c, I-d, and I-e, the series of ortho-SeCl-substituted benzoyl chlorides can react with different 3-amino-2,6-piperidinedione or 3-amino-1,4-dihydropyridine-2-(1H)-one or 3-amino-1-adamantanol or 2-benzothiazolamine or 3-amino-2,5-pyrroledione respectively to afford the corresponding desired products of formulas I-a, I-b, I-c, I-d, while using the series of ortho-I or Br-substituted of benzoyl chlorides as the starting substrates will obtain the o-halobenzamide intermediates firstly, which can react with [Se] reagents furtherly to produce the desired benzoisoselazolidone derivatives of formulas I-a, I-b, I-c, I-d and I-e; in the second synthesis route for preparing the selenium-containing isoxazolamines represented by general formula I-a, I-b, I-c, I-d, I-e, and I-f, the different substituted 2,2′-diselenylbisbenzaldehydes can respectively react with different 3-amino-2,6-piperidinedione or 3-amino-1,4-dihydropyridine-2-(1H)-one or 3-amino-1-adamantanol or 2-benzothiazolamine or 3-amino-2,5-pyrroledione to afford the corresponding imine intermediates, which can produce the desired selenium-containing isoxazolamines via the reductive amination; in the third synthesis route for preparing the tetravalent selenium-containing isoxazolamines represented by general formula I-a, I-b, I-c, I-d, I-e, and I-f, the benzisoselenidazole derivatives can be direct oxidated to the desired products with [O.sup.−] reagents.

6. A method of treating a disease of autoimmune diseases, neurological degenerative diseases, hematological tumors, solid tumors, myelofibrosis, and acute/chronic graft-versus-host response which are caused by the overexpression of TNF-α, comprising administering to a subject a therapeutically or prophylactically effective amount of a selenium-containing isoxazolamine derivative of claim 1, a pharmaceutically acceptable salt, a solvate, a polymorph, a stereoisomer, an isotopic compound, or a metabolite thereof.

7. A method of treating a disease of neurological degenerative diseases, hematological tumors, solid tumors, tissue ischemia-reperfusion injury, acute renal failure, and aging diseases which are caused by abnormal ferroptosis-like cell death, comprising administering to a subject a therapeutically or prophylactically effective amount of a selenium-containing isoxazolamine derivative of claim 1, a pharmaceutically acceptable salt, a solvate, a polymorph, a stereoisomer, an isotopic compound, or a metabolite thereof.

8. A method of treating a disease, symptom or disorder caused by the overexpression of TNF-α and/or abnormal ferroptosis-like cell death, wherein the method comprises administering to a subject a therapeutically or prophylactically effective amount of a substance selected from the group consisting of general formula (I) and the pharmaceutically acceptable salt thereof according to claim 1.

9. (canceled)

10. The method according to claim 7, wherein the autoimmune diseases include myelofibrosis, acute/chronic graft-versus-host response disease, rheumatoid arthritis, inflammatory bowel disease, diabetes, psoriasis, mandatory spondylitis, leprosy nodular erythema, and other infectious diseases such as HBV, HCV, HIV; the neurodegenerative diseases include Alzheimer's disease, dementia, multiple sclerosis, motor neuron disease; the blood tumor refers to multiple bone marrow tumor, myelodysplastic syndrome; the solid tumor refers to liver cancer, kidney cancer, gastric cancer, colon cancer, ovarian cancer, pancreatic cancer, prostate cancer, breast cancer, melanoma, and cerebral glioblastoma; the tissue ischemic reperfusion injury refers to stroke, coronary heart disease, myocardial infarction, pulmonary embolism, and acute coronary syndrome.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0063] FIG. 1 is the general structure of. the compounds of the present invention according to formula I.

[0064] FIG. 2 is the protective effect of the compound of formula I on ox-LDL-induced human vascular endothelial cell injury.

[0065] FIG. 3 is the protective effect of the compound of formula I on erastin-induced ferroptosis-like cell death in HT22 cells.

EXAMPLES

[0066] The present invention will now be further elucidated by way of a description of a preferred exemplary embodiment of the invention, but is not limited thereto. Each preferred conditions aforementioned can be combined randomly without departing from the common knowledge in the art thereby forming various preferred embodiments of the present invention.

[0067] In the following embodiments, 1H-NMR was measured with a Varian Mercury AMX300 instrument. MS was measured with VG ZAB-HS or VG-7070 and Esquire 3000Plus-01005. All reaction solvents are redistilled before use, and the anhydrous solvents are obtained in accordance with standard drying methods. Unless otherwise indicated, all reactions were carried out under the protection of argon and monitored by TLC, and following the conventional workup and pre-drying treatment by saturated saline and anhydrous sodium sulfate. Products were purified by column chromatography on silica gel (200-300 mesh) unless otherwise stated.

Example 1. Synthesis of Compound 1

[0068] Step 1: Preparation of 2-Chloroselenobenzoyl Chloride

[0069] Anthranilic acid (1.37 g, 10 mmol) was added to a 3N aqueous hydrochloric acid solution (4 ml) under an ice bath, and then sodium nitrite (10 mmol, 690 mg) in an aqueous solution (2 ml) was slowly added to the reaction mixture under stirring, and the reaction will change to a clear solution after an hour and to give the desired 2-benzoic acid diazonium salt solution.

[0070] Selenium powder (790 mg, 10 mmol) was mixed with cetyltrimethyl ammonium bromide (20 mg) and 2N aqueous sodium hydroxide solution (5 ml) to obtain a Se—NaOH solution under nitrogen atmosphere. NaBH.sub.4 (49 mg, 1.3 mmol) solution (1 mL) that contained NaOH (40 mg, 1 mmol) was added drop-wisely into the Se—NaOH solution under an ice bath. The reaction was stirred at room temperature for 1 h and then at 90° C. for 0.5 h to from Na.sub.2Se.sub.2. After cooling to the room temperature, the 2-benzoic acid diazonium salt solution obtained above was slowly added dropwise to the Na.sub.2Se.sub.2 solution, and the mixture was heated to 40° C. for 2 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was acidified by adding 6N HCl until the precipitate was no longer precipitated, filtered, the filter cake was washed with water and dried over to give a khaki solid of 2,2′-disselenized bisbenzoic acid in 80% yield, mp 295-296° C.

[0071] 2,2′-Diselenized bisbenzoic acid (800 mg, 2 mmol) was added to a sulfoxide solution (5 ml), and the mixture was heated to reflux for 3 hours under nitrogen atmosphere. The excessive sulfoxide was removed by evaporation under vacuum, while the residue was extracted with anhydrous n-hexane. The combined organic phases were evaporated under vacuum to give a yellow solid, which could be recrystallized from diethyl ether to obtain 2-chloroselenobenzoyl chloride as a pale-yellow solid with a yield of 81%, mp 60-62° C.

[0072] Step 2: Synthesis of Target Compound 1

##STR00018##

[0073] A solution of dichloroselenobenzoyl chloride (254 mg, 1 mmol) in acetonitrile (2 mL) was added dropwise to a stirred acetonitrile solution (10 mL) of 3-amino-2,6-piperidinedione (128 mg, 1 mmol) and triethylamine (151 mg, 1.5 mmol) under nitrogen atmosphere and ice bath (J. Med. Chem. 2016, 59, 8125-8133). After the reaction was completed (monitored by TLC), 20 ml of water was added, followed by extraction with ethyl acetate (20 mL×2). The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated under reduced pressure and purified by silica gel column chromatography. (V.sub.acetone:V.sub.petroleum ether=1:4 to 1:1) to give compound 1 (260 mg, yield 85%). HRMS-ESI: m/z calcd for C.sub.12H.sub.10N.sub.2O.sub.3Se: 309.9857, found [M+H].sup.+ 310.9927; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.98 (s, 1H), 8.07 (d, J=8.0 Hz, 1H), 7.85 (d, J=7.4 Hz, 1H), 7.68-7.62 (m, 1H), 7.45 (t, J=7.4 Hz, 1H), 5.26 (dd, J=12.8, 5.3 Hz, 1H), 2.92-2.83 (m, 1H), 2.59 (d, J=17.3 Hz, 1H), 2.48-2.36 (m, 1H), 2.13-2.05 (m, 1H); .sup.13C NMR (126 MHz, DMSO) δ 173.2, 171.3, 167.5, 140.4, 132.25, 128.0, 127.9, 126.3, 126.2, 53.7, 31.7, 25.1.

[0074] Example 2 to 14 were performed according to the operation of example 1, wherein the synthesis of substituted 2-chloroselenobenzoyl chloride could follow the reference methods (J. Med. Chem. 2016, 59, 8125-8133 or Bioorg Med Chem. 2012, 20, 3816-3827) that using 2-aminobenzoic acid as the raw material (commercially available) and following by diselenyl etherification and chlorination reaction. The compounds listed in examples 15 to 16 were synthesized according to the above route and using 6-aminopenicilanic acid instead of 3-amino-2,6-piperidinedione; the compounds listed in examples 17 to 21 were synthesized according to the above route and using 3-amino-2,5-pyrroledione instead of 3-amino-2,6-piperidinedione. Examples of results obtained are as follows:

TABLE-US-00001 Examples Compound information 2 Compound 2, molecular formula: C.sub.12H.sub.9N.sub.3O.sub.5Se, yield 48%. HRMS-ESI: m/z 354.9707, found [M + H].sup.+ 355.9781. 3 Compound 4, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.3Se, yiled 72%. HRMS-ESI: m/z 327.9762, found [M + H].sup.+ 328.9835; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.02 (s, 1H), 8.02-7.95 (m, 1H), 7.58-7.47 (m, 2H), 5.32-5.23 (m, 1H), 2.96-2.87 (m, 1H), 2.69-2.56 (m, 1H), 2.49-2.36 (m, 1H), 2.15- 2.03 (m, 1H). 4 Compound 5, molecular formula: C.sub.12H.sub.9N.sub.3O.sub.5Se, yiled 75%. HRMS-ESI: m/z 354.9707, found [M + H].sup.+ 355.9785; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.11 (s, 1H), 8.57-8.40 (m, 2H), 7.87-7.70 (m, 1H), 5.32-5.23 (m, 1H), 2.96-2.87 (m, 1H), 2.68-2.58 (m, 1H), 2.49-2.37 (m, 1H), 2.15- 2.03 (m, 1H). 5 Compound 7, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.3Se, yiled 73%. HRMS-ESI: m/z 327.9762, found [M + H].sup.+ 328.9837; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.01 (s, 1H), 8.04 (dd, J = 8.6, 5.3 Hz, 1H), 7.30-7.27 (m, 1H), 7.17 (dt, 8.6, 2.4 Hz, 1H), 5.30-5.23 (m, 1H), 2.96-2.87 (m, 1H), 2.64-2.55 (m, 1H), 2.49-2.36 (m, 1H), 2.17-2.02 (m, 1H). 6 Compound 8, molecular formula: C.sub.12H.sub.9ClN.sub.2O.sub.3Se, yiled 62%. HRMS-ESI: m/z 343.9467, found [M + H].sup.+ 344.9540; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.03 (s, 1H), 8.01 (dd, J = 8.2, 3.5 Hz, 1H), 7.62-7.38 (m, 2H), 5.30-5.24 (m, 1H), 2.97-2.88 (m, 1H), 2.66-2.53 (m, 1H), 2.47-2.36 (m, 1H), 2.17-2.02 (m, 1H). 7 Compound 9, molecular formula: C.sub.14H.sub.13N.sub.3O.sub.4Se, yiled 58%. MS-ESI [M + H].sup.+ 368.1. 8 Compound 10, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.3Se, yiled 52%. MS-ESI [M + H].sup.+ 329.0. 9 Compound 11, molecular formula: C.sub.13H.sub.12N.sub.2O.sub.4Se, yiled 79%. HRMS-ESI: m/z 339.9962, found [M + H].sup.+ 341.0035; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.81 (s, 1H), 7.95 (d, J = 7.8 Hz, 1H), 7.31 (d, J = 7.5 Hz, 1H), 7.20 (t, J = 7.8 Hz, 1H), 5.24-4.73 (m, 1H), 3.92 (s, 3H), 2.95-2.83 (m, 1H), 2.65- 2.53 (m, 1H), 2.48-2.32 (m, 1H), 2.12-1.95 (m, 1H). 10 Compound 12, molecular formula: C.sub.13H.sub.12N.sub.2O.sub.4Se, yiled 67%. HRMS-ESI: m/z 339.9962, found [M + H].sup.+ 341.0033; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.95 (s, 1H), 7.85 (d, J = 8.8 Hz, 1H), 7.29-7.20 (m, 2H), 5.21 (dd, J = 12.8, 5.3 Hz, 1H), 3.82 (s, 3H), 2.92-2.83 (m, 1H), 2.59 (d, J = 17.3 Hz, 1H), 2.48-2.37 (m, 1H), 2.12-2.02 (m, 1H). 11 Compound 13, molecular formula: C.sub.14H.sub.14N.sub.2O.sub.5Se, yiled 83%. HRMS-ESI: m/z 370.0068, found [M + H].sup.+ 371.0142; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.96 (s, 1H), 7.49 (s, 1H), 7.06 (s, 1H), 5.19 (dd, J = 12.8, 5.3 Hz, 1H), 3.97 (s, 3H), 3.96 (s, 3H), 2.92-2.82 (m, 1H), 2.62-2.57 (m. 1H), 2.48- 2.38 (m, 1H), 2.13-2.01 (m, 1H). 12 Compound 14, molecular formula: C.sub.15H.sub.16N.sub.2O.sub.6Se, yiled 65%. HRMS-ESI: m/z 400.0174, found [M + H].sup.+ 401.0246; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.97 (s, 1H), 7.12 (s, 1H), 5.20 (dd, J = 12.8, 5.2 Hz, 1H), 3.97 (s, 3H), 3.96 (s, 3H), 3.95 (s, 3H), 2.91-2.83 (m, 1H), 2.56 (d, J = 17.3 Hz, 1H), 2.47-2.33 (m, 1H), 2.03-1.92 (m, 1H). 13 Compound 15, molecular formula: C.sub.13H.sub.12N.sub.2O.sub.3Se, yiled 87%. HRMS-ESI: m/z 324.0013, found [M + H].sup.+ 325.0085; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.89 (s, 1H), 7.95 (d, J = 7.8 Hz, 1H), 7.41-7.27 (m, 2H), 5.24-4.73 (m, 1H), 2.88-2.43 (m, 3H), 2.34 (s, 3H), 2.08-1.92 (m, 1H). 14 Compound 16, molecular formula: C.sub.13H.sub.11N.sub.3O.sub.4Se, yiled 55%. MS-ESI [M + H].sup.+ 354.0. 15 Compound 17, molecular formula: C.sub.15H.sub.14N.sub.2O.sub.4SSe. HRMS-ESI: m/z 397.9839, found [M + H].sup.+ 398.9912; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.11 (d, J = 7.8 Hz, 1H), 7.88-7.63 (m, 2H), 7.47 (t, J = 7.4 Hz, 1H), 5.98 (s, 1H), 5.71 (s, 1H), 4.47 (s, 1H), 1.71 (s, 3H), 1.53 (s, 3H). 16 Compound 18, molecular formula: C.sub.16H.sub.14FNO.sub.4SSe. HRMS-ESI: m/z 415.9745, found [M + H].sup.+ 416.9818. 17 Compound 24, molecular formula: C.sub.11H.sub.8N.sub.2O.sub.3Se. HRMS-ESI: m/z 295.9700, found [M + H].sup.+ 296.9773; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.41 (s, 1H), 8.06 (d, J = 8.1 Hz, 1H), 7.84-7.62 (m, 2H), 7.43 (t, J = 7.4 Hz, 1H), 5.30 (dd, J = 9.8, 5.7 Hz, 1H), 2.98 (dd, J = 18.0, 9.8 Hz, 1H), 2.86 (dd, J = 18.0, 5.7 Hz, 1H). 18 Compound 25, molecular formula: C.sub.12H.sub.10N.sub.2O.sub.4Se. HRMS-ESI: m/z 325.9806, found [M + H].sup.+ 326.9879; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.36 (s, 1H), 7.90 (d, J = 8.6 Hz, 3H), 7.36-7.23 (m, 1H), 5.27 (dd, J = 9.8, 5.7 Hz, 1H), 3.87 (s, 3H), 2.95-2.82 (m, 2H). 19 Compound 26, molecular formula: C.sub.11H.sub.7FN.sub.2O.sub.3Se. HRMS-ESI: m/z 313.9606, found [M + H].sup.+ 314.9682; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.41 (s, 1H), 8.06 (d, J = 8.1 Hz, 1H), 7.84-7.62 (m, 2H), 7.43 (t, J = 7.4 Hz, 1H), 5.30 (dd, J = 9.8, 5.7 Hz, 1H), 2.98 (dd, J = 18.0, 9.8 Hz, 1H), 2.86 (dd, J = 18.0, 5.7 Hz, 1H). 20 Compound 27, molecular formula: C.sub.11H.sub.7FN.sub.2O.sub.3Se. HRMS-ESI: m/z 313.9606, found [M + H].sup.+ 314.9682; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.42 (s, 1H), 8.07 (dd, J = 8.6, 5.2 Hz, 1H), 7.42- 7.31 (m, 2H), 5.33-5.28 (m, 1H), 2.98-2.83 (m, 2H). 21 Compound 28, molecular formula: C.sub.12H.sub.10N.sub.2O.sub.3Se. HRMS-ESI: m/z 309.9857, found [M + H].sup.+ 310.9930; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.29 (s, 1H), 8.01 (d, J = 7.8 Hz, 1H), 7.37-7.23 (m, 1H), 5.33-5.25 (m, 1H), 2.96-2.81 (m, 2H), 2.35 (s, 3H).

Example 22. Synthesis of Compound 19

[0075] Synthesis Route:

##STR00019##

[0076] Step 1: Synthesis of Intermediate a19

[0077] To a stirred solution of 3-amino-1-adamantanol (167 mg, 1 mmol) and triethylamine (151 mg, 1.5 mmol) in tetrahydrofuran (10 mL) was slowly added o-iodobenzoyl chloride (266 mg, 1 mmol) under ice bath, and the resulting mixture was reacted for 1 to 2 h. After the reaction was completed (monitored by TLC), 20 ml of water was added, followed by extraction with ethyl acetate (20 mL×2). The combined organic phases were sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated under reduced pressure and purified by silica gel column chromatography (V.sub.ethyl acetate:V.sub.petroleum ether=1:4 to 1:1) to give the key intermediate a19. MS-ESI [M+H].sup.+ 398.0.

[0078] Step 2: Synthesis of Compound 19

[0079] To a stirred solution of a19 (397 mg, 1 mmol), selenium powder (0.15 g, 1.9 mmol), and K.sub.2CO.sub.3(276 mg, 2 mmol) in DMF (5 mL) were added CuI (154 mg, 0.8 mmol), 1,10-o-diphenyl azaphenanthrene (146 mg, 0.8 mmol), and the resulting mixture was reacted at 110° C. for 24 hours under nitrogen atmosphere. After the reaction was completed (monitored by TLC), 20 ml of water was added, followed by extraction with ethyl acetate (20 mL×2). The combined organic phases were sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated under reduced pressure and purified by silica gel column chromatography (V.sub.acetone:V.sub.petroleum ether=1:4 to 1:1) to give compound 19 (251 mg, yield 72%). HRMS-ESI: m/z C.sub.17H.sub.19NO.sub.2Se: 349.0581, found [M+H].sup.+ 350.0655; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.04 (d, J=8.2 Hz, 1H), 7.83 (d, J=7.4 Hz, 1H), 7.68-7.60 (m, 1H), 7.43 (t, J=7.4 Hz, 1H), 2.48-1.60 (m, 15H).

[0080] Example 23 to 29 were performed according to the operation of example 22, wherein examples 24 to 26 were synthesized using 2-amino-benzothiophene instead of 3-amino-1-amantadine, examples of results obtained are as follows:

TABLE-US-00002 Examples Compound information 23 Compound 20, molecular formula: C.sub.17H.sub.18FNO.sub.2Se, [M + H].sup.+ 368.1. 24 Compound 21, molecular formula: C.sub.14H.sub.8N.sub.2OSSe, HRMS-ESI: m/z 331.9523, found [M + H].sup.+ 332.9595; .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.16 (dd, J = 7.4, 3.1 Hz, 1H), 8.07-7.92 (m, 5H), 7.57 (t, J = 7.4 Hz, 1H), 7.48 (t, J = 7.4 Hz, 1H). 25 Compound 22, molecular formula: C.sub.15H.sub.7F.sub.3N.sub.2O.sub.2SSe, HRMS-ESI: m/z 415.9346, found [M + H].sup.+ 416.9418; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.20 (s, 1H), 8.15 (d, J = 8.1 Hz, 1H), 8.04 (d, J = 7.4 Hz, 1H), 7.96 (d, J = 8.8 Hz, 1H), 7.79 (t, J = 7.5 Hz, 1H), 7.55 (t, J = 7.4 Hz, 1H), 7.48-7.38 (m, 1H). 26 Compound 23, molecular formula: C.sub.15H.sub.10N.sub.2O.sub.2SSe, MS-ESI [M + H].sup.+ 363.0. 27 Compound 29, molecular formula: C.sub.15H.sub.10N.sub.2OSSe, MS-ESI [M + H].sup.+ 346.9. 28 Compound 30, molecular formula: C.sub.16H.sub.9F.sub.3N.sub.2O.sub.3SSe, MS-ESI [M + H].sup.+ 446.9. 29 Compound 31, molecular formula: C.sub.15H.sub.10FN.sub.2O.sub.2SSe, MS-ESI [M + H].sup.+ 380.9.

Example 30. Synthesis of Compound 3

[0081] To a solution of compound 2 (72 mg, 0.2 mmol) in methanol (1 mL) was added 10% Pd/C, and the resulting mixture was heated to 70° C. under H.sub.2 atmosphere for 48 hours. After the reaction was completed (monitored by TLC), the mixture was filtered, the obtained filtrate was evaporated under reduced pressure and purified by column chromatography to give compound 6 (25 mg, 36%). HRMS-ESI: m/z calcd for C.sub.12H.sub.10N.sub.2O.sub.3Se: 324.9966, found [M+H].sup.+ 326.0040; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.97 (s, 1H), 7.47-7.34 (m, 1H), 7.05-6.83 (m, 2H), 6.33 (brs, 2H), 5.23 (dd, J=12.8, 5.3 Hz, 1H), 2.92-2.83 (m, 1H), 2.59 (d, J=17.3 Hz, 1H), 2.45-2.35 (m, 1H), 2.12-2.02 (m, 1H).

Example 31. Synthesis of Compound 6

[0082] To a solution of compound 5 (72 mg, 0.2 mmol) in methanol (1 mL) was added 10% Pd/C and 80% hydrazine hydrate (40 mg) under ice bath and nitrogen atmosphere, and the reaction was heated to 40° C. for 8 hours. After the reaction was completed (monitored by TLC), the mixture was filtered, the obtained filtrate was evaporated under reduced pressure and purified by column chromatography to give compound 6 (15 mg, 22%). HRMS-ESI: m/z calcd for C.sub.12H.sub.11N.sub.3O.sub.3Se: 324.9966, found [M+H].sup.+ 326.0040; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.97 (s, 1H), 7.29-6.91 (m, 3H), 6.12 (brs, 2H), 5.21 (dd, J=12.8, 5.3 Hz, 1H), 2.94-2.82 (m, 1H), 2.59-2.52 (m, 1H), 2.43-2.29 (m, 1H), 2.12-2.01 (in, 1H).

Example 32. Synthesis of Compound 32

[0083] To a stirred solution of compound 19 (35 mg, 0.1 mmol) in methanol (2 ml) was slowly added 30% hydrogen peroxide (0.12 mmol) under ice bath, the reaction temperature was raised to room temperature and performed for 12 hours. After the reaction was completed (monitored by TLC), 5 ml of water was added, and then ethyl acetate (5 mL×2) was added for extraction. The organic phase was sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was evaporated under reduced pressure and purified by silica gel column chromatography (V.sub.ethyl acetate:V.sub.petroleum ether=1:1) to produce compound 32 (27 mg, yield 75%). HRMS-ESI: m/z calcd for C.sub.17H.sub.19NO.sub.3Se: 365.0530, found [M+H].sup.+ 366.0609; .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.24-7.96 (m, 2H), 7.83-7.62 (m, 2H), 2.76-1.71 (n, 15H).

[0084] Example 33 to 42 were performed according to the operation of example 32, examples of results obtained are as follows:

TABLE-US-00003 Examples Compound information 33 Compound 33, yiled 26%, molecular formula: C.sub.14H.sub.8N.sub.2O.sub.2SSe, HRMS-ESI: m/z 347.9472, found [M + H].sup.+ 348.9548; .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.18-7.95 (m, 6H), 7.62 (t, J = 7.1 Hz, 1H), 7.52 (t, J = 7.1 Hz, 1H). 34 Compound 34, yiled 21%, molecular formula: C.sub.15H.sub.7F.sub.3N.sub.2O.sub.3SSe, MS-ESI [M + H].sup.+ 432.9. 35 Compound 36, yiled 43%, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.4Se, MS-ESI [M + H].sup.+ 345.0. 36 Compound 37, yiled 47%, molecular formula: C.sub.14H.sub.13N.sub.3O.sub.5Se, MS-ESI [M + H].sup.+ 384.0. 37 Compound 38, yiled 35%, molecular formula: C.sub.13H.sub.9N.sub.3O.sub.4Se, MS-ESI [M + H].sup.+ 352.0. 38 Compound 39, yiled 29%, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.4Se, HRMS-ESI: m/z 343.9712, found [M + H].sup.+ 344.9784; .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.08 (s, 1H), 8.12-8.08 (m, 1H), 7.42-7.23 (m, 2H), 5.31-5.27 (m, 1H), 2.96- 2.87 (m, 1H), 2.64-2.53 (m, 1H), 2.48-2.35 (m, 1H), 2.17-2.02 (m, 1H). 39 Compound 40, yiled 51%, molecular formula: C.sub.13H.sub.12N.sub.2O.sub.5Se, MS-ESI [M + H].sup.+ 357.0. 40 Compound 41, yiled 39%, molecular formula: C.sub.11H.sub.8N.sub.2O.sub.4Se, HRMS-ESI: m/z 311.9649, found [M + H].sup.+ 312.9724; .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.49 (s, 1H), 8.28-7.82 (m, 3H), 7.59 (t, J = 7.4 Hz, 1H), 5.33-5.27 (m, 1H), 3.02-2.95 (m, 1H), 2.89-2.82(m, 1H). 41 Compound 42, yiled 42%, molecular formula: C.sub.12H.sub.10N.sub.2O.sub.4Se, MS-ESI [M + H].sup.+ 327.0. 42 Compound 43, yiled 34%, molecular formula: C.sub.12H.sub.10N.sub.2O.sub.5Se, MS-ESI [M + H].sup.+ 343.0.

Example 43. Synthesis of Compound 35

[0085] ##STR00020##

[0086] Compound a5 could be obtained through a peroxidation according to the synthesis method described in example 32. To a stirred solution of compound a5 (37 mg, 0.1 mmol) in THF (1 mL) was slowly added an aqueous solution (0.7 ml) of sodium bisulfite (52 mg, 0.5 mmol) under nitrogen atmosphere, and the reaction was performed at 60° C. After the reaction was completed (monitored by TLC), the reaction mixture was cooled to room temperature, and extracted with ethyl acetate (5 mL×2). The combined organic phases were sequentially washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The obtained filtrate was evaporated under reduced pressure to give compound 35 (27 mg, yield 79%). Molecular formula: C.sub.12H.sub.11N.sub.3O.sub.4Se, HRMS-ESI: m/z calcd for 340.9915, found [M+H].sup.+ 341.9984.

Example 44. Synthesis of Compound 44

Step 1: Preparation of 2,2′-Diselenylbisbenzaldehyde

[0087] 3-Methoxy-2-aminobenzaldehyde (1.51 g, 10 mmol) was added to a 3N hydrochloric acid water-DMSO (V:V=1:1) mixed solution (4 mL) under an ice bath, and then sodium nitrite (10 mmol, 690 mg) in an aqueous solution (2 ml) was slowly added to the reaction mixture under stirring, and the reaction will change to a clear solution after an hour and to give the desired diazonium salt solution.

[0088] Selenium powder (790 mg, 10 mmol) was mixed with cetyltrimethyl ammonium bromide (20 mg) and 2N aqueous sodium hydroxide solution (5 ml) to obtain a Se—NaOH solution under nitrogen atmosphere. NaBH.sub.4 (49 mg, 1.3 mmol) solution (1 mL) that contained NaOH (40 mg, 1 mmol) was added drop-wisely into the Se—NaOH solution under an ice bath. The reaction was stirred at room temperature for 1 h and then at 90° C. for 0.5 h to from Na.sub.2Se.sub.2. After cooling to the room temperature, the diazonium salt solution obtained above was slowly added dropwise to the Na.sub.2Se.sub.2 solution, and the mixture was heated to 40° C. for 2 hours. After the reaction was completed, the reaction mixture was acidified to neutral by adding 1N HCl, and then extracted with ethyl acetate for two times, the combined organic phases were evaporated under reduced pressure and purified by silica gel column chromatography (eluted with dichloromethane and methanol) to produce a khaki solid of 2,2′-diselenylbisbenzaldehyde in 45% yield. Molecular formula: C.sub.16H.sub.14O.sub.4Se.sub.2, HRMS-ESI: m/z calcd for 428.9223, found [M+H].sup.+ 430.9298; .sup.1H NMR (CDCl.sub.3), δ (ppm): 3.78 (s, 6H), 7.09-7.13 (dd, J=7.6 Hz, 2H), 7.42-7.50 (m, 4H), 10.20 (s, 2H).

Step 2: Synthesis of Target Compound 44

[0089] ##STR00021##

[0090] To a stirred solution of 2,2′-diselenylbisbenzaldehyde (429 mg, 1 mmol) in 20 ml of acetonitrile was added 50 μL of conc HCl and stirred for 15 min, then 3-amino-2,6-piperidinedione (128 mg, 1 mmol) was added and stirred for 6 hours to obtain the enamine intermediate. The above reaction mixture was concentrated under reduced pressure and redissolved with 10 ml of methanol, followed by adding sodium borohydride (38 mg, 1 mmol) under ice bath and the reaction was performed for another 6 h. After the reaction is completed, the resulting mixture was extracted with ethyl acetate, dried over magnesium sulfate, filtered, concentrated and purified by silica gel column chromatography to give compound 44 in 36% yield, 117 mg. HRMS-ESI: m/z calcd for C.sub.13H.sub.14N.sub.2O.sub.3Se: 326.0170, found [M+H].sup.+ 327.0245; .sup.1H NMR (DMSO-d.sub.6), δ (ppm): 10.91 (s, 1H), 7.85-7.69 (t, J=7.8 Hz, 1H), 7.58-7.31 (m, 2H), 4.75-4.62 (m, 3H), 3.89 (s, 3H), 2.78-2.42 (m, 2H), 2.31-1.91 (m, 2H).

[0091] Examples 45 to 48 were performed according to the operation of example 44, examples of results obtained are as follows:

TABLE-US-00004 Examples Compound information 45 Compound 48, yiled 23%, molecular formula: C.sub.16H.sub.11F.sub.3N2O.sub.2SSe, HRMS-ESI: m/z 431.9659, found [M + H].sup.+ 432.9730; .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.06-7.35 (m, 6H), 4.86 (s, 2 H), 3.92 (s, 3 H). 46 Compound 49, yiled 41%, molecular formula: C.sub.16H.sub.18F.sub.3N.sub.2O.sub.4SSe, MS-ESI [M + H].sup.+ 415.0. 47 Compound 50, yiled 49%, molecular formula: C.sub.12H.sub.9FN.sub.2O.sub.4Se, HRMS-ESI: m/z 365.0894, found [M + H].sup.+ 366.0967; .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.21-7.15 (t, J = 7.8 Hz, 1H), 6.93-6.73 (m, 1H), 4.40 (s, 2H), 3.84 (s, 1H), 2.43-1.51 (m, 15H). 48 Compound 51, yiled 34%, molecular formula: C.sub.13H.sub.12N.sub.2O.sub.5Se, HRMS-ESI: m/z 312.0013, found [M + H].sup.+ 313.0086.

Effect Examples

Example 49. TNF-α Activity Inhibiting Assay

[0092] Methods: Peripheral blood from healthy volunteers was collected with EDTA anticoagulant tubes. After being diluted 5-fold with 1640 medium (Gibco, USA), the blood was added to 96-well cell culture plates (Costar, USA) and then treated with 10 μL solution of the compound of general formula (I) of the present invention in DMSO (Sigma, USA), and the final concentration of DMSO was 0.2%. After incubation for 60 minutes in an incubator at 37° C. under 5% CO.sub.2, 10 μL LPS (Sigma, USA) was added to the reaction system, and the final concentration was 10 ng/mL. After further culturing for 6 hours in the incubator at 37° C. under 5% CO.sub.2, the supernatant was collected. The content of TNF-α was determined by ELISA (BD Biosciences, USA). Absorbance was detected at OD.sub.450 nm with a microplate reader, with GD 650 nm as reference. The control, a solution containing 0.2% DMSO medium, was as 0% inhibition. Raw data and standard curves were recorded. The four-parameter drug inhibition curve was plotted by XL-fit software and the inhibition rate of each compound was calculated. The experimental results are shown in table 1.

TABLE-US-00005 TABLE 1 TNF-α inhibitory activity Comps TNF-α inhibition (%) 1 C 2 B 3 A 4 C 5 B 6 A 7 C 8 C 9 C 10 B 11 B 12 B 13 B 14 A 15 A 16 B 17 D 18 D 19 C 20 C 21 C 22 B 23 C 24 C 25 C 26 C 27 B 28 A 29 B 30 C 31 D 32 D 33 D 34 C 35 A 36 C 37 C 38 A 39 C 40 B 41 C 44 B 50 C Thalidomide D Lenadomide A Note: A: <1 μM; B: 1~10 μM; C: 10~100 μM; D: >100 μM.

Example 50 Thioredoxin Reductase 1 (TrxR1) Activity Inhibiting Assay

[0093] Working solutions: 0.119 mg/mL of TrxR working solution preparation: 175 μL of TrxR stock solution (0.34 mg/mL) was diluted to 500 μL; 1 mM NADPH working solution preparation, NADPH (5 mg) was dissolved in 12 mL potassium phosphate buffer; 1 mM DTNB working solution preparation: 25 mg DTNB was dissolve in 63 mL DMSO; potassium phosphate buffer system preparation: 0.2 mg/mL bovine serum albumin (BSA) and 1 mM EDTA were added to potassium phosphate buffer at pH 7.4 (K.sub.2HPO.sub.4/KH.sub.2PO.sub.4).

[0094] Methods: The TrxR1 inhibitory activity study of selenium-containing isoxazolamines in vitro. Insulin, NADPH, Trx, the samples and TrxR1 were added to a microcuvette in order, and subsequently added the work buffer (0.1 mol/L potassium phosphate/2 mmol/L EDTA) to a total volume of 0.5 mL. The concentration of each component in the obtained reaction system is: insulin 130 μmol/L, NADPH 0.4 mol/L (Sigma), Trx 4 μmol/L, the added substrate (the selenium-containing isoxazolamines treated samples). After determination of the initial absorption at 340 nm, the enzymatic reaction was started by adding 20 μg extracted TrxR1 proteins, the decrease in absorbance at 340 nm was monitored. One unit of enzymatic activity is defined as the consumption. The activity can be calculated from the extinction coefficient of NADPH, and the unit of enzyme activity is defined as: 1U=ΔA340 nm/min×1000, the TR activity could be calculated with U/L. The results are shown in table 2.

TABLE-US-00006 TABLE 2 TrxR inhibition activity of compounds of formula I Comps TrxR inhibition IC.sub.50 1 B 2 B 4 A 5 C 7 A 8 B 9 B 10 A 12 A 13 A 16 B 17 A 21 B 22 A 23 B 25 B 27 B 30 A 31 B 34 C 38 D 40 C 45 B Ebselen C Note: A: <5 μM; B: 5~50 μM; C: 50~100 μM; D: >100 μM.

Example 51: Glutathione Peroxidase (GPx)-Like Activity Assay

[0095] Method: GPX activity of selenium compounds was assayed by ultraviolet spectrophotometry. Glutathione (2.0 mM), EDTA (1 mM), glutathione disulfide reductase (1.7 units mL.sup.−1), and nicotinamide adenine dinucleotide phosphate oxidase (NADPH; 0.4 mM) were mixed intopotassium phosphate buffer (0.1 M, pH=7.5). The selenium compounds (50 μm) was added to the above mixture At room temperature (25° C.), and the reaction was started by the addition of H.sub.2O.sub.2, tBuOOH or Cum-OOH (1.6 mM) respectively. The initial reduction rates were calculated from the rate of NADPH oxidation at 340 nm in a GSH assay. Each initial rate was measured at least three times and calculated from the first 5-10% of the reaction by using 6.22 mM.sup.−1 cm.sup.−1 as the molar extinction coefficient for NADPH. For the peroxidase activity, the rates were corrected for the background reaction between peroxide and thiol. The results are shown in table 3.

TABLE-US-00007 TABLE 3 Anti-peroxidation effects of selenium-containing isoxazolamines represented as formula I Initial rate v.sub.0 [μm min.sup.−1] Comps H.sub.2O.sub.2 tBuOOH Cum-OOH 1 109.7 ± 2.1 42.0 ± 3.3 124.5 ± 7.6  7 128.2 ± 2.0 49.3 ± 5.0 142.2 ± 8.2  12 121.7 ± 2.2 54.5 ± 5.7 116.8 ± 7.2  19  99.7 ± 1.9 32.3 ± 3.5 34.2 ± 1.5 21 115.3 ± 2.6 35.7 ± 4.2 94.5 ± 4.2 26 145.2 ± 3.4 55.7 ± 5.2 164.5 ± 9.1  32  36.7 ± 0.8 12.0 ± 2.3 11.5 ± 0.2 34  39.7 ± 1.4 14.2 ± 1.1 12.4 ± 0.3 Ebselen  76.7 ± 1.1 21.0 ± 4.3 31.2 ± 0.5
The results showed that some of the selenium-containing isoxazolamines had better anti-peroxidation activities than the positive control ebselen.

Example 52 Effect on Endothelial Cell Damage Caused by Ox-LDL

[0096] Drugs: Selenium-containing isoxazolamines were dissolved in DMSO.

[0097] Reagents: Ox-LDL (Beijing Xiehe Sanyou); DMEM medium (Low sugar, GIBCO, UK); HMEC cells (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences); MTT (Sigma, product No. 5655, USA); other reagents are analytical grade.

[0098] Method: HMEC cells were cultured in DMEM media supplemented with 10% FBS at 37° C. with 5% CO.sub.2. The MTT assay was used for the determination of cytotoxicity. The cells were harvested at logarithmic phase and plated at a density of 2×10.sup.4 cells per well in 96-well plates, cultured for 48 hours and grew into a tightly packed single molecular layer. Using the serum-free DMEM medium instead, and 100 μL aliquots of medium containing 5 μM of the selenium-containing isoxazolamines or contrast were added subsequently. After incubating for 1 hour, 100 μg/ml ox-LDL was added to the injured groups. After 24 h of incubation, cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. In brief, MTT (100 μL, 0.5 mg mL.sup.−1) was added to the wells for 4 h at 37° C., discarded the culture medium, DMSO 150 μL/well was added and shaked for 5 minutes to release the dye, and measured the OD at 570 nm with a microplate reader. The changes in OD caused by cytokine and drug treatment were used as an index of cell viability, normalised to cells incubated in control medium, which were considered 100% viable.

[0099] Results: As shown in FIG. 2, ox-LDL could cause HMEC cells damage, and the selenium-containing isoxazolamines could significantly inhibit HMEC cells death caused by ox-LDL.

Example 53. Effect of the Selenium-Containing Isoxazolamines on Erastin-Induced Ferroptosis in HT22 Cells

[0100] Drugs: Selenium-containing isoxazolamines and erastin, which could be dissolved by DMSO.

[0101] Reagents: CCK-8 kit (Sigma, USA), DEME medium (Sigma, USA), mouse HT22 hippocampal cells (Shanghai Jiaotong University).

[0102] Method: HT22 cells were cultured in DMEM media supplemented with 10% fetal bovin serum at 37° C. with 5% CO.sub.2. The culture HT22 cells were plated in 96-well plates and allowed to incubate for 24 hours. Subsequently, 100 μL aliquots of medium containing 5 μM of the selenium-containing isoxazolamines was added and cultured for 2 hours. Afterward, 0.5 μmol/L Erastin was added and cultured for 8 hours, and then added 10 μL of the CCK-8 solution per well and incubated for 3 hours, and measured the OD at 450 nm with a microplate reader. The cell survival rate was calculated according to the following formula: cell survival rate %=(treatment group-blank control group)/(control group-blank control group)*100%. The experiment was repeated three times.

[0103] Results: As shown in FIG. 3, erastin could cause apoptosis in HT22 cells, and the selenium-containing isoxazolamines could significantly reduce the erastin-induced ferroptosis and improve cell survival rate.