THIADIAZOLE-SUBSTITUTED COMPOUND, PHARMACEUTICAL COMPOSITIONS THEREOF, AND APPLICATIONS THEREOF

Abstract

Disclosed is a thiadiazole-substituted compound, pharmaceutical composition thereof and application thereof. The present disclosure provides a compound containing structure of a thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof. The thiadiazole-substituted compound has strong inhibitory effect on PARG and and better pharmacokinetics, is expected to treat and/or prevent various PARG-related diseases.

Claims

1. A thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof: ##STR00019## wherein, represents a single bond or a double bond; B is CF.sub.3 or CHF.sub.2; A.sub.1 is N or CR.sup.b1, R.sup.b is hydrogen or halogen; and when B is CHF.sub.2, A.sub.1 is CR.sup.b1, R.sup.b1 is halogen; A.sub.2 is N, C or CR.sup.c1; R.sup.c1 is hydrogen, halogen, C.sub.1-6 alkyl or hydroxyl; A.sub.3 is CR.sup.23 or CHR.sup.23; R.sup.a1 is cyano or C.sub.1-6 alkyl; R.sup.21 and R.sup.22 are independently hydrogen or C.sub.1-6 alkyl, and when R.sup.21 is hydrogen, R.sup.22 is C.sub.1-6 alkyl; R.sup.23 is hydrogen, halogen, hydroxyl, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkyl substituted with one or more R.sup.23-1, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, OC.sub.1-6 alkyl, SC.sub.1-6 alkyl, C(O)R.sup.2a, NR.sup.2b1R.sup.2b2, C(O)OR.sup.2c, C(O)NR.sup.2d1R.sup.2d2, C.sub.3-10 cycloalkyl, 4- to 12-membered heterocycloalkyl containing 1 to 3 heteroatoms independently selected from O, S and N, C.sub.6-20 aryl, 5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S and N, C.sub.4-8 cycloalkenyl, or, 4- to 8-membered heterocycloalkenyl containing 1 to 3 heteroatoms independently selected from O, S and N; R.sup.23-1 is independently halogen, cyano, C.sub.1-6 alkyl-O, hydroxyl, C(O)R.sup.23a, NR.sup.23b1R.sup.23b2, C(O)OR.sup.23e or C(O)NR.sup.23d1R.sup.23d2; R.sup.2a, R.sup.2b1, R.sup.2b2, R.sup.2c, R.sup.2d1, R.sup.2d2, R.sup.23a, R.sup.23b1, R.sup.23b2, R.sup.23c, R.sup.23d1 and R.sup.23d2 are independently hydrogen, C.sub.1-6 alkyl, or, C.sub.1-6 alkyl substituted with one or more halogen; or, R.sup.2b1 and R.sup.2b2, R.sup.2d1 and R.sup.2d2, R.sup.23b1 and R.sup.23b2, R.sup.23d1 and R.sup.23d2 independently together with the N to which they are attached form 3- to 8-membered heterocycle containing 1 to 3 heteroatoms, wherein, one heteroatom is N, other heteroatoms, if any, independently selected from O, S and N, or 3- to 8-membered heterocycle containing 1 to 3 heteroatoms, wherein, one heteroatom is N, other heteroatoms, if any, independently selected from O, S and N substituted with one or more R.sup.1-2-1; R.sup.1-2-1 is independently independently halogen, C.sub.1-6 alkyl, or, C.sub.1-6 alkyl substituted with one or more halogen.

2. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, represents a single bond or a double bond; B is CF.sub.3 or CHF.sub.2; A.sub.1 is N or CR.sup.b1, R.sup.b1 is hydrogen or halogen, and when B is CHF.sub.2, A.sub.1 is CR.sup.b1, R.sup.b1 is halogen; A.sub.2 is N or C; A.sub.3 is CR.sup.23 or CHR.sup.23; R.sup.23 is hydrogen; R.sup.a1 is cyano or C.sub.1-6 alkyl; R.sup.21 and R.sup.22 are independently hydrogen or C.sub.1-6 alkyl, and when R.sup.21 is hydrogen, R.sup.22 is C.sub.1-6 alkyl.

3. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, A.sub.2 is N or C; and/or, A.sub.3 is CR.sup.23 or CHR.sup.23; R.sup.23 is hydrogen; and/or, when R.sup.21 and R.sup.22 are independently C.sub.1-6 alkyl, ##STR00020## wherein, ##STR00021## represents R conformation, S conformation or a mixture of R and S conformation; and/or, when the definition of R.sup.b1, R.sup.c1, R.sup.23, R.sup.23-1, R.sup.2a, R.sup.2b1, R.sup.2b2, R.sup.2c, R.sup.2d1, R.sup.2d2, R.sup.23a, R.sup.23b1, R.sup.23b2, R.sup.23c, R.sup.23d1, R.sup.23d2 and R.sup.1-2-1 refers to halogen, the halogen is fluorine, chlorine, bromine or iodine; and/or, when the definition of R.sup.c1, R.sup.a1, R.sup.21, R.sup.22, R.sup.23, R.sup.2a, R.sup.2b1, R.sup.2b2, R.sup.2c, R.sup.2d1, R.sup.2d2, R.sup.23a, R.sup.23b1, R.sup.23b2, R.sup.23c, R.sup.23d1, R.sup.23d2 and R.sup.1-2-1 refers to C.sub.1-6 alkyl, the C.sub.1-6 alkyl is C.sub.1-4 alkyl.

4. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, A.sub.1 is CF or CH; and/or, is a single bond, A.sub.2 is N, A.sub.3 is CH.sub.2, R.sup.21 and R.sup.22 are independently C.sub.1-6 alkyl; and/or, R.sup.a1 is cyano or methyl; and/or, R.sup.21 and R.sup.22 are independently hydrogen or methyl, and when R.sup.21 is hydrogen, R.sup.22 is methyl.

5. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, the thiadiazole-substituted compound represented by formula I, having the structure represented by formula II: ##STR00022## wherein, B is CF.sub.3 or CHF.sub.2; R.sup.b1 is hydrogen or halogen, and when B is CHF.sub.2, R.sup.b1 is halogen; R.sup.a1 is cyano or C.sub.1-6 alkyl; R.sup.21 and R.sup.22 are independently C.sub.1-6 alkyl.

6. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, ##STR00023##

7. The thiadiazole-substituted compound represented by formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an isomer thereof or an isotopic compound thereof according to claim 1, wherein, the thiadiazole-substituted compound represented by formula I is any one of the following structures: ##STR00024## ##STR00025## ##STR00026## ##STR00027## ##STR00028## ##STR00029##

8. A pharmaceutical composition comprising a substance A and a pharmaceutically acceptable excipient, wherein the substance A is a therapeutically effective amount of the thiadiazole-substituted compound represented by formula I, the pharmaceutically acceptable salt thereof, the stereoisomer thereof, the tautomer thereof or the isotopically labeled compound thereof according to claim 1.

9. A method of inhibiting PARG in a subject in need thereof, comprising: administering a therapeutically effective amount of a substance A to the subject, wherein the substance A is the thiadiazole-substituted compound represented by formula I, the pharmaceutically acceptable salt thereof, the stereoisomer thereof, the tautomer thereof or the isotopically labeled compound thereof according to claim 1.

10. A method of treating or preventing a PARG related disease in a subject in need thereof, comprising: administering an effective amount of a substance A, wherein the substance A is the thiadiazole-substituted compound represented by formula I, the pharmaceutically acceptable salt thereof, the stereoisomer thereof, the tautomer thereof or the isotopically labeled compound thereof according to claim 1.

11. In the method for treating or preventing an PARG related disease in a subject in need thereof according to claim 10, wherein the PARG related disease is cancer, the cancer is selected from the group consisting of colon cancer, appendicle cancer, pancreatic cancer, MYH-related polyposis, hematologic cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophageal cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia and melanoma.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0065] The present disclosure is further illustrated by the following examples, which are not intended to limit the present disclosure. Experimental procedures without specified conditions in the following examples were performed in accordance with conventional procedures and conditions, or in accordance with instructions. The solvents involved in the following embodiments are analytically or chromatographically pure. When the solvents involved in the following embodiments are mixed solvents, they are all volume ratios unless otherwise stated.

[0066] The following is a list of abbreviations used in the embodiments: [0067] DCM dichloromethane [0068] DMF N, N-dimethylformamide [0069] DMSO Dimethyl sulfoxide [0070] DIPEA diisopropylethylamine [0071] DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene [0072] NMP N-methylpyrrolidone [0073] Pd.sub.2(dba).sub.3 tris(dibenzylideneacetone) dipalladium [0074] Xantphos 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene [0075] SelectFluor II I-Fluoro-4-methyl-1,4-diazabicyclo[2.2.2]octane-1,4-diium tetrafluoroborate

Example 1 Synthetic Route of Compound 1

##STR00013##

Synthesis of Compounds 1-c

[0076] 2-Bromo-5-(trifluoromethyl)-1,3,4-thiadiazole (678 mg, 2.91 mmol) and anhydrous DMF (8 mL) were added into a microwave tube. 6-Bromo-4-fluoro-1H-indazole (500 mg, 2.33 mmol) and cesium carbonate (1820 mg, 5.59 mmol) were added at room temperature and sealed. The reaction mixture was heated and stirred at 60 C. for 2 hours. The reaction mixture was cooled to room temperature, added into ice water, acidified by adding solid sodium bisulfate and extracted with ethyl acetate twice. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated to dryness by rotary evaporation, and dried by an oil pump to give 1-c (852 mg, 100%).

Synthesis of Compound 1-b

[0077] In a reaction vial 1-c (750 mg, 2.04 mmol), Pd.sub.2(dba).sub.3 (90 mg, 0.098 mmol), XANT PHOS (90 mg, 0.16 mmol), DIPEA (792 mg, 6.13 mmol), 1,4-dioxane (20 mL), and benzyl mercaptan (0.31 mL, 2.66 mmol). After degassed and purged with nitrogen for three times, the reaction mixture was heated and stirred at 90 C. for 1.5 hours, monitored, it was not finished, then stirred at 90 C. for another 1.5 hours. Cooled to room temperature, the reaction mixture was removed solvent by rotary evaporation, and the residue was added ethyl acetate and petroleum ether, washed with water, brine, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate, 100/0 to 90/10) to obtain compound 1-b (800 mg, 95% yield). LC-MS (ESI): m/z 411.1 (M+H).sup.+.

Synthesis of Compound 1-a

[0078] To a reaction flask were added 1-b (800 mg, 1.95 mmol), acetonitrile (20 mL), acetic acid (0.13 mL, 2.33 mmol) and water (0.14 mL, 7.77 mmol). Dichlorhydantoin (768 mg, 3.90 mmol) was added to the above mixture in an ice-water bath. The reaction mixture was stirred in an ice-water bath for 1 h. The solvent was removed by rotary evaporation at room temperature. The residue was added ethyl acetate and petroleum ether, washed with water, brine, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and dried by an oil pump to obtain the crude product. The crude product was dissolved in anhydrous dichloromethane (10 mL) to obtain solution A. To a reaction flask were added 1-methylcyclopropanamine hydrochloride (420 mg, 3.90 mmol), triethylamine (1.36 mL, 9.75 mmol) and anhydrous dichloromethane (7 mL) in an ice-water bath. The reaction mixture was stirred for 5 min in an ice-water bath and then was added Solution A, and anhydrous dichloromethane (4 mL) washed the flask that had contained the Solution A twice and added into the above reaction mixture. The reaction mixture was stirred at room temperature for 1 h, then was washed with water, brine, dried over anhydrous sodium sulfate, filtered, evaporated, and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate, 100/0 to 80/20) to give compound 1-a (330 mg, 40% yield). LC-MS (ESI): m/z 422.0 (M+H).sup.+.

Synthesis of Compound 1

[0079] 1-a (90 mg, 0.21 mmol) and (2S,6S)-2,6-dimethylpiperazine dihydrochloride (120 mg, 0.64 mmol) were added to a reaction flask. The mixture was degassed to vacuum and added anhydrous NMP (4 mL). After degassed and purged with nitrogen for 3 times, the above mixture was added DBU (195 mg, 1.28 mmol). The reaction mixture was heated and stirred at 80 C. for 6 hours. The reaction mixture was cooled to room temperature, added water, extracted with ethyl acetate twice, dried over anhydrous sodium sulfate, filtered, evaporated, and purified by column chromatography (C8 column, mobile phase: 10 mM ammonium bicarbonate, acetonitrile) to obtain the crude (95% purity), and then purified on a silica column again (mobile phase: dichloromethane/methanol, 100/0 to 94/6) to obtain compound 1 (61 mg, 55% yield). LC-MS (ESI): m/z 516.2 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHZ) 8.80 (1H, s), 8.39 (1H, s), 8.34 (1H, s), 7.13 (1H, s), 3.43-3.37 (4H, m), 3.11 (2H, dd, J=12.0, 6.4 Hz), 1.19 (6H, d, J=6.4 Hz), 1.08 (3H, s), 0.71-0.61 (2H, m), 0.45-0.36 (2H, m).

Example 2 Synthetic Route of Compound 2

##STR00014##

Synthesis of Compound 2-a

[0080] To a reaction flask were added 1-b (270 mg, 0.66 mmol), acetonitrile (11 mL), acetic acid (0.043 mL, 0.75 mmol) and water (0.045 mL, 2.50 mmol). Dichlorhydantoin (266 mg, 1.35 mmol) was added to the above mixture in an ice-water bath. The reaction mixture was stirred in an ice-water bath for 1 hour and concentrated to dryness by rotary evaporation at low temperature.

[0081] The residue was added ethyl acetate and petroleum ether, washed with water, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and dried by an oil pump for 10 min. added dichloromethane (5 mL) to give Solution A. To a reaction flask was added 1-amino-1-cyclopropyl cyanide hydrochloride (312 mg, 2.63 mmol), and to this flask were added pyridine (0.5 mL), dichloromethane (6 mL) and the Solution A sequentially under an ice-water bath. The reaction mixture was stirred at room temperature for 3 h. Sodium bisulfate aqueous solution was added. Aqueous sodium bisulfate was added, partitioned, and the organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude was purified by column chromatography (mobile phase: dichloromethane/methanol, 100/0 to 95/5) to afford compound 2-a (170 mg, 60% yield). LC-MS (ESI): m/z 433.1 (M+H).sup.+.

Synthesis of Compound 2

[0082] 2-a (70 mg, 0.162 mmol) and (2S,6S)-2,6-dimethylpiperazine dihydrochloride (90.6 mg, 0.484 mmol) were added to a reaction vial. The above mixture was degassed to vacuum and was added anhydrous NMP (2 mL). Degassed and purged with nitrogen twice and DBU (138 mg, 0.906 mmol) was added. The reaction mixture was heated and stirred at 80 C. for 6 hours, then was cooled to room temperature, added water and extracted twice with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated, and the residue was purified by column chromatography (mobile phase: dichloromethane/(dichloromethane/methanol/ammonia 90:10:0.2), 100/0 to 35/65), and lyophilized in acetonitrile diluted with ammonia to give compound 2 (51 mg, 60% yield). LC-MS (ESI): m/z 527.1 (M+H).sup.+. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 8.84 (1H, s), 8.42 (1H, s), 7.14 (1H, s), 3.42 (2H, dd, J=11.6, 2.4 Hz), 3.37-3.34 (2H, m), 3.12 (2H, dd, J=11.6, 6.4 Hz), 1.47-1.39 (2H, m), 1.36-1.26 (2H, m), 1.19 (6H, d, J=6.4 Hz).

Example 3 Synthetic Route of Compound 3

##STR00015##

Synthesis of Compound 3

[0083] Compound 2 (50 mg, 0.095 mmol) and SelectFluor II (91 mg, 0.29 mmol) were combined in a reaction flask. After degassed to vacuum, the above mixture was added anhydrous acetonitrile (5 mL). Degassed and purged with nitrogen for 2 times, the above mixture was added acetic acid (0.027 mL, 0.47 mmol). The reaction mixture was stirred at 40 C. for 4.5 hours, then was cooled to room temperature, added ethyl acetate, pure water and anhydrous sodium sulfate, partitioned, and the aqueous phase was extracted with ethyl acetate once. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated and purified by prep-HPLC (ammonium bicarbonate) to give compound 3 (11.2 mg, 22% yield). LC-MS (ESI): m/z 545.7 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 8.94 (1H, s), 8.51 (1H, d, J=4.4 Hz), 3.53-3.49 (4H, m), 3.17 (2H, dd, J=12.0, 6.0 Hz), 1.46-1.37 (2H, m), 1.34-1.27 (2H, m), 1.20 (6H, d, J=6.4 Hz).

Example 4 Synthetic Route of Compound 4

##STR00016##

Synthesis of Compound 4

[0084] Compound 1 (70 mg, 0.14 mmol) and SelectFluor II (130 mg, 0.41 mmol) were added to a reaction flask. After degassed to vacuum, the mixture was added anhydrous acetonitrile (5 mL), then degassed and purged with nitrogen for 2 times, the mixture was added acetic acid (0.029 mL, 0.50 mmol). The reaction mixture was stirred at room temperature for 10 hours, then was added ethyl acetate, pure water and anhydrous sodium sulfate, partitioned and the aqueous phase was extracted with ethyl acetate once. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated and purified by prep-HPLC (ammonium bicarbonate) to give compound 4 (19.2 mg, 28% yield). LC-MS (ESI): m/z 534.3 (M+H).sup.+. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 8.91 (1H, d, J=0.4 Hz), 8.59 (1H, brs), 8.45 (1H, d, J=4.8 Hz), 3.50-3.41 (2H, m), 3.31-3.22 (2H, m), 3.10 (2H, dd, J=11.2, 6.0 Hz), 1.17-1.12 (9H, m), 0.74-0.65 (2H, m), 0.47-0.41 (2H, m).

Synthesis of Comparative Compound 1

##STR00017##

[0085] Synthesized according to the synthetic method of patent WO2023183850A1.

Example 5 Synthetic Route of Compound 5

##STR00018##

Synthesis of Compound 5

[0086] Compound 1 (160 mg, 0.32 mmol), 1-fluoro-4-methyl-1,4-diazabicyclo[2.2.2]octane tetrafluoroborate (309 mg, 0.97 mmol), and acetic acid (525 mg, 8.74 mmol) were added to acetonitrile (10 mL), and the reaction mixture was stirred for 10 hr at room temperature after the reaction mixture was degassed and purged with nitrogen for three times. The pH was adjusted to 78 by adding saturated sodium bicarbonate solution to the reaction mixture, to which water (50 mL) was added and extracted with ethyl acetate (50 mL). The organic phase was washed with brine (50 mL), dried over sodium sulfate, filtered off the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (mobile phase, DCM/MeOH 20/1), and then purified by a C18 column (NH.sub.4HCO.sub.3) to obtain compound 5 (24 mg, 14%). LC-MS (ESI): m/z 516.4 (M+H).sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6): 8.89 (1H, s), 8.60 (1H, s), 8.49 (1H, d, J=4.0 Hz), 7.61 (1H, t, J=52.0 Hz), 3.55-3.46 (2H, m), 3.43-3.36 (2H, m), 3.20-3.11 (2H, m), 1.25-1.18 (7H, m), 1.14 (3H, s), 0.75-0.67 (2H, m), 0.50-0.40 (2H, m).

Effectiveness Data

Bioactivity Test

PARG Enzyme Inhibition Assay

Experimental Procedure.

[0087] PARG in vitro assays were conducted in standard 384-well plates in a total volume of 15 L. 5 L of PARG (389-976) (manufactured by Chempartner Chemical Co., Ltd.) in buffer (50 mM Tris-HCL 7.5, 30 mM KCl, 1 mM EDTA, 3 mM DTT, tween-20 0.01%, BSA 0.025%) was added at a final concentration of 1.5 pM to the 384-well plates containing the compounds to be tested, which was incubated for 30 min at room temperature. To the above mixture was added 5 L Bio PARylated His-TEV-PARP1 (2-1014) substrate (manufactured by Chempartner Chemical Co., Ltd.) at a final concentration of 12 nM, after addition, the resulting mixture was incubated for 30 minutes at room temperature. Then to the mixture was added detection reagent (5 L) which was buffered with 50 mM Tris-HCL 7.5, 30 mM KCl, 1 mM EDTA, 3 mM DTT, tween-20 0.01%, BSA 0.025%, and consisted of 3 M of compound PDD00017273 and 9 nM Mab anti-6HIS XL665 (Cisbio: 61HISXLA) and 0.9 nM streptavidin affinity terbium cryptate (Cisbio: 610SATLA), all at 3 working concentrations (final concentrations of 1 M, 3 nM and 0.3 nM, respectively). After 120 min incubation in the dark at room temperature, TR-FRET signals were measured at Ex 340 and Em 665 and Em 615. The ratio for each well was calculated as Em 665/Em 615 and the compound inhibition rate was calculated based on the obtained data.

PARG Biochemical Activity of Representative Compounds of the Present Disclosure

TABLE-US-00001 IC.sub.50 IC.sub.50 IC.sub.50 Compound (PARG Compound (PARG Compound (PARG No. enzyme) No. enzyme) No. enzyme) 1 ***** 2 ***** 1 ***** (0.20 nM) (0.62 nM) (0.21 nM) 4 ***** 5 ***** ***** represents IC.sub.50 < 2 nM; **** represents 2 nM IC.sub.50 < 10 nM; *** represents 10 nM IC.sub.50 < 100 nM; ** represents 100 nM IC.sub.50 < 1 M; * represents IC.sub.50 1 M
Pharmacokinetic Studies in Mice after Intravenous and Oral Administration of the Drug Experimental Procedure:

[0088] After intravenous and oral administration to male C57BL/6J mice or male CD-1 mice, plasma samples were collected through the orbital venous plexus at 8-9 time points (e.g., 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 7 h, 24 h or 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h). The collected blood samples were transferred to microcentrifuge tubes containing EDTA-K2 anticoagulant, and the supernatants were centrifuged for 5 min at 4 C. and 4000 g. The supernatants were stored in a refrigerator at 75 C.15 C. The concentrations of the compounds in the plasma at the different time points were detected by LC-MS/MS and the relevant pharmacokinetic parameters were calculated using the WinNonlin software

Results of Pharmacokinetic Testing of Representative Compounds of the Present Disclosure

TABLE-US-00002 Test compounds compound Compound Compound way of administration 1.sup.b 1.sup.b 3.sup.b intravenous.sup.a1 Cl 22.5 89.5 34.8 (5 mg/kg) (mL/min/kg) AUC.sub.0-last 2539 905 2407 (h*ng/mL) AUC.sub.0-inf 3703 939 2417 (h*ng/mL) oral C.sub.max (h) 175 98.8 513 administration.sup.a2 AUC.sub.0-last 2608 725 4069 (10 mg/kg) (h*ng/mL) AUC.sub.0-inf 2975 929 4085 (h*ng/mL) way of administration compound 4.sup.b compound 5.sup.b intravenous.sup.a1 Cl 35.6 51.6 (5 mg/kg) (mL/min/kg) AUC.sub.0-last 1987 1603 (h*ng/mL) AUC.sub.0-inf 2348 1617 (h*ng/mL) oral C.sub.max (h) 198 172 administration.sup.a2 AUC.sub.0-last 3260 1974 (10 mg/kg) (h*ng/mL) AUC.sub.0-inf 3973 1998 (h*ng/mL) .sup.bmale CD-1 mice, mean value of blood samples analyzed separately .sup.a1Intravenous drug delivery solvents is 5% DMSO + 10% Solutol HS-15 + 85% (20% HP--CD in water); .sup.a2oral administration drug delivery solvents is 0.1% Tween80 + 0.5% MC in water.

[0089] The results showed that the compounds of the present disclosure possessed significant advantages in PK parameters relative to the comparative compound, with a significant increase in the exposure level.

[0090] Although specific embodiments of the present disclosure have been described above, it will be appreciated by those skilled in the art that these embodiments are merely illustrative and that many changes or modifications can be made to these embodiments without departing from the principles and spirit of the present disclosure. The scope of protection of the present disclosure is therefore defined by the appended claims.