Substituted boric acid compound, pharmaceutical composition comprising same, and application thereof

Abstract

A substituted boric acid compound, a pharmaceutical composition including the same, and an application thereof. The substituted boric acid compound is a compound represented by formula (I), or a crystal form, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, a hydrate, or a solvent compound thereof. The boric acid compound has proteasome inhibitory activity, good pharmacodynamic/pharmacokinetic performance, good applicability, and high safety, and can be used for preparing drugs for treating diseases related to proteasomes. ##STR00001##

Claims

1. A compound represented by Formula (I), or a crystalline form, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, a hydrate or a solvate thereof, ##STR00019## wherein R.sup.4, R.sup.5, R.sup.6, R.sup.7, and R.sup.8 are each independently hydrogen, deuterium, or halogen; R.sup.1, R.sup.2, R.sup.3, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, and R.sup.15 are hydrogen; provided that at least one of R.sup.4, R.sup.5, R.sup.6, R.sup.7, and R.sup.8 is deuterium.

2. The compound according to claim 1, wherein R.sup.4 and R.sup.5 are deuterium.

3. The compound according to claim 1, wherein R.sup.6 is deuterium.

4. The compound according to claim 2, wherein R.sup.6 is deuterium.

5. The compound according to claim 1, wherein R.sup.7 and R.sup.8 are deuterium.

6. The compound according to claim 2, wherein R.sup.7 and R.sup.8 are deuterium.

7. The compound according to claim 3, wherein R.sup.7 and R.sup.8 are deuterium.

8. The compound according to claim 4, wherein R.sup.7 and R.sup.8 are deuterium.

9. The compound according to claim 1, wherein the compound is selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof: ##STR00020##

10. The compound according to claim 1, wherein the compound is: ##STR00021## or a pharmaceutically acceptable salt thereof.

11. The compound according to claim 1, wherein the compound is: ##STR00022## or a pharmaceutically acceptable salt thereof.

12. The compound according to claim 1, wherein the compound is: ##STR00023## or a pharmaceutically acceptable salt thereof.

13. The compound according to claim 1, wherein the compound is: ##STR00024## or a pharmaceutically acceptable salt thereof.

14. The compound according to claim 1, wherein the compound is: ##STR00025## or a pharmaceutically acceptable salt thereof.

15. A pharmaceutical composition, comprising a pharmaceutically acceptable carrier, and a compound according to claim 1, or a crystalline form, a pharmaceutically acceptable salt, a hydrate, a solvate, a stereoisomer, or a prodrug.

16. A pharmaceutical composition, comprising a pharmaceutically acceptable carrier, and a compound according to claim 9, or a pharmaceutically acceptable salt thereof.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

(1) The methods of preparing the compound of Formula (I) disclosed herein are more specifically described below, but these specific methods do not constitute any limitation to the present invention. The compounds disclosed herein may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations can be readily made by those skilled in the art to which the present invention pertains.

Example 1 Preparation of (R)-(1-(2-(2,5-dichlorobenzamido)-2,2-d2-acetamido)-3-methylbutyl)boronic acid (Compound I-1)

(2) ##STR00007##

(3) The specific synthetic steps are as follows:

(4) ##STR00008## ##STR00009##

Step 1: Synthesis of 2,2-d2-glycine (Compound 3)

(5) Glycine (200 mg, 2.66 mmol) and salicylaldehyde (0.04 mL, 0.376 mmol) were added to a reaction flask, dissolved by adding 10 mL of deuterated acetic acid, heated to 100° C. under nitrogen, and stirred for 2 hours. The reaction was cooled to room temperature, and concentrated to remove acetic acid. Then, 2 mL of heavy water was added, and stirred at room temperature for 15 minutes. The mixture was diluted in a small amount of water, decolorized for 0.5 hour with activated carbon, filtered, and the filtrate was concentrated to dryness. A small amount of methanol was added, and a white solid was precipitated under stirring, which was filtered, and dried in vacuo to give a product (176.7 mg, yield: 88%). LC-MS (APCI): m/z=78.1 (M+1).sup.+.

Step 2: Synthesis of 2,5-[(dichlorobenzoyl)amino]-d2-acetic Acid (Compound 5)

(6) The compound 3 (131 mg, 1.7 mmol) and sodium hydroxide (212.5 mg, 5.31 mmol) were added to a reaction flask, and dissolved by adding 2 mL of water. A solution of 2,5-dichlorobenzoyl chloride (176.7 mg, 0.85 mmol) in 1 mL tetrahydrofuran was added dropwise in an ice bath, and the reaction was stirred for 1 hour after the addition. After TLC detected the reaction was completed, the pH of the reaction was adjusted to acidic with a dilute hydrochloric acid, and a white solid was precipitated, which was filtered, washed with ice water, and dried in vacuo to give a product (200 mg, yield: 94.5%). LC-MS (APCI): m/z=250.3 (M+1).sup.+.

Step 3: Synthesis of 2-methylpropylboronic acid-(+)-pinanediol Ester (Compound 8)

(7) Isobutylboronic acid (510.5 mg, 5 mmol) and (1S,2S,3R,5S)-(+)-2,3-pinanediol (850.65 mg, 5 mmol) were added to a reaction flask, and dissolved by adding 5 mL of n-heptane. The reaction was stirred at room temperature for 2 hours. After TLC detected the reaction was completed, the mixture was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and dried in vacuo to give a product (1.1 g, yield: 93.14%).

Step 4: Synthesis of (S)-1-chloro-3-methylbutylboronic acid-(+)-pinanediol Ester (Compound 9)

(8) The compound 8 (1.1 g, 4.66 mmol) was added to a reaction flask, and dissolved by adding 6.5 mL of anhydrous tetrahydrofuran and 1.54 mL of anhydrous dichloromethane. The temperature was lowered to −78° C. under nitrogen, and a 2 M solution of lithium diisopropylamide (LDA, 4.66 mL, 9.32 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 2 hours. A 1 M solution of zinc chloride in tetrahydrofuran (8.16 mL, 8.16 mmol) was slowly added dropwise. After the addition, the reaction was reacted at the low temperature for 2 hours. After TLC detected the reaction was completed, a small amount of 10% dilute sulfuric acid was added to quench the reaction, and the organic phase was separated. The aqueous phase was extracted with n-hexane, and the organic phases were combined, washed with brine, and concentrated. The residue was purified by silica gel column chromatography and dried in vacuo to give a product (1.31 g, yield: 98.9%). LC-MS (APCI): m/z=285.5 (M+1).sup.+.

Step 5: Synthesis of (R)-1-(hexamethyldisilanyl)amino-3-methylbutylboronic Acid Pinanediol Ester (Compound 10)

(9) The compound 9 (1.35 g, 4.75 mmol) was added to a reaction flask, dissolved by adding 10 mL of anhydrous tetrahydrofuran, and the temperature was lowered to −40° C. under nitrogen. Lithium bis(trimethylsilyl)amide (LiHMDS, 5.7 mL, 5.7 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 1 hour, and then the reaction was warmed to room temperature and reacted for 1 hour. After TLC detected the reaction was completed, the mixture was filtered through a plug of silica gel, and the filtrate was evaporated to dryness to give a product (1.9 g, yield: 97.7%), which was used directly in the next step.

Step 6: Synthesis of L-leucine Boronic Acid-(+)-pinanediol Ester Trifluoroacetate Salt (Compound 11)

(10) The compound 10 (1.7 g, 4.15 mmol) was added to a dry reaction flask, dissolved by adding 6 mL of diisopropyl ether, and the temperature was lowered to −15° C. under nitrogen.

(11) Trifluoroacetic acid (TFA, 1.89 g, 16.6 mmol) was slowly added dropwise. After the addition, the reaction was kept at this temperature and reacted for 2 hours, and then allowed to warm to room temperature and reacted overnight. The mixture was filtered, and the filter cake was washed with diisopropyl ether and dried in vacuo to give a white solid (620 mg, yield: 39.5%). LC-MS (APCI): m/z=266.2 (M+1).sup.+.

Step 7: Synthesis of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylene-1,3,2-benzodioxaborolan-2-yl]butyl}amino)-2-oxa-1,1-d2-ethyl]benzamide (Compound 12)

(12) The compound 5 (100 mg, 0.402 mmol) and the compound 11 (152.3 mg, 0.402 mmol) were added to a reaction flask, O-benzotriazol-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU, 142 mg, 0.44 mmol) was added under nitrogen, and the substances were dissolved by adding 2 ml of anhydrous N,N-dimethylformamide (DMF), and N,N-diisopropylethylamine (DIPEA, 156 mg, 1.21 mmol) was added under ice bath. After the addition, the reaction was warmed to room temperature and stirred for 1 hour. After TLC detected the reaction was completed, the mixture was diluted with a small amount of water and extracted 3-4 times with EtOAc. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography to give a product (196 mg, yield: 98.2%). LC-MS (APCI): m/z=497.5 (M+1).sup.+.

Step 8: Synthesis of (R)-(1-(2-(2,5-dichlorobenzamido)-2,2-d2-acetamido)-3-methylbutyl)boronic Acid (Compound I-1)

(13) The compound 12 (196 mg, 0.395 mmol) and isobutylboronic acid (104.8 mg, 1.03 mmol) were added to a reaction flask, and dissolved by adding 2 mL of methanol and 2 mL of n-hexane, 1N hydrochloric acid (0.5 mL, 0.5 mmol) was then added, and the reaction was stirred at room temperature under nitrogen protection overnight. After TLC detected the reaction was completed, the methanol layer was separated, washed three times with n-heptane, concentrated to dryness under reduced pressure, dissolved by adding 2N sodium hydroxide and washed three times with dichloromethane. The aqueous phase was adjusted to pH 2-3 with concentrated hydrochloric acid and extracted with dichloromethane for 3-4 times. The organic phases were combined, washed with brine, concentrated, and purified by silica gel column chromatography. The product was dried and weighed 85 mg (yield: 59.4%). LC-MS (APCI): m/z=345.2 (M+1-H.sub.2O). .sup.1H NMR (400 MHz, DMSO) δ 8.93 (s, 1H), 8.71 (d, J=7.8 Hz, 1H), 7.65 (s, 1H), 7.55 (d, J=1.3 Hz, 2H), 2.64 (s, 1H), 1.61 (dd, J=13.3, 6.5 Hz, 1H), 1.40-1.32 (m, 1H), 1.26 (s, 1H), 0.85-0.80 (m, 6H).

Example 2 Preparation of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-1-d-3-methylbutyl)boronic Acid (Compound I-2)

(14) ##STR00010##

(15) The specific synthetic steps are as follows:

(16) ##STR00011##

Step 1: Synthesis of 2,5-[(dichlorobenzoyl)amino]acetic Acid (Compound 13)

(17) Glycine (1.125 g, 15 mmol) and sodium hydroxide (750 mg, 18.75 mmol) were added to a reaction flask, and dissolved by adding 7.5 mL of water. A solution of 2,5-dichlorobenzoyl chloride (623.7 mg, 3.0 mmol) in 1 mL of tetrahydrofuran was added dropwise in an ice bath. After the addition, the reaction was stirred for 1 hour. After TLC detected the reaction was completed, the pH was adjusted to acidic with dilute hydrochloric acid, and a white solid was precipitated, which was filtered, washed with ice water and dried in vacuo to give a product (675 mg, yield: 91.1%). LC-MS (APCI): m/z=248.3 (M+1).sup.+.

Step 2: Synthesis of (S)-1-chloro-1-d-3-methylbutylboronic acid-(+)-pinanediol Ester (Compound 14)

(18) The compound 8 (1.1 g, 4.66 mmol) was added to a reaction flask, and dissolved by adding 6.5 mL of anhydrous tetrahydrofuran (THF) and 1.54 mL of deuterated dichloromethane. The temperature was lowered to −78° C. under nitrogen protection, and a 2 M solution of LDA (4.66 mL, 9.32 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 2 h. A 1 M solution of zinc chloride in tetrahydrofuran (8.16 mL, 8.16 mmol) was slowly added dropwise. After the addition, the reaction was continued for 2 hours at the low temperature. After TLC detected the reaction was completed, a small amount of 10% dilute sulfuric acid was added to quench the reaction, and the organic phase was separated. The aqueous phase was extracted with n-hexane, and the organic phases were combined, washed with brine, and concentrated. The residue was purified by silica gel column chromatography and dried in vacuo to give a product (1.29 g, yield: 97.0%). LC-MS (APCI): m/z=286.2 (M+1).sup.+.

Step 3: Synthesis of (R)-1-(hexamethyldisilanyl)amino-1-d-3-methylbutylboronic Acid Pinanediol Ester (Compound 15)

(19) The compound 14 (1.29 g, 4.52 mmol) was added to a reaction flask, dissolved by adding 9 mL of anhydrous tetrahydrofuran, and the temperature was lowered to −40° C. under nitrogen protection. Lithium bis(trimethylsilyl)amide (5.4 mL, 5.4 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 1 hour, and then warmed to room temperature and reacted for 1 hour. After TLC detected the reaction was completed, the mixture was filtered through a plug of silica gel, and the filtrate was evaporated to dryness to give a product (1.6 g, yield: 86.3%), which was directly used in the next step.

Step 4: Synthesis of 1-d-L-leucine boronic acid-(+)-pinanediol Ester Trifluoroacetate Salt (Compound 16)

(20) The compound 15 (1.6 g, 3.8 mmol) was added to a dry reaction flask, dissolved by adding 6 mL of diisopropyl ether, and the temperature was lowered to −15° C. under nitrogen protection. Trifluoroacetic acid (1.75 g, 15.4 mmol) was slowly added dropwise. After the addition, the reaction was reacted at this temperature for 2 hours and then allowed to warm to room temperature and reacted overnight. After filtration, the filter cake was washed with diisopropyl ether and dried in vacuo to give a white solid (561 mg, yield: 38.9%). LC-MS (APCI): m/z=267.7 (M+1).sup.+.

Step 5: Synthesis of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylene-1,3,2-benzodioxaborolan-2-yl]-1-d-butyl}amino)-2-oxaethyl]benzamide (Compound) 17)

(21) The compound 16 (100 mg, 0.405 mmol) and the compound 13 (154 mg, 0.405 mmol) were added to a reaction flask, TBTU (143 mg, 0.46 mmol) was added under nitrogen protection, and dissolved by adding 2 mL of anhydrous DMF. DIPEA (157 mg, 1.21 mmol) was added dropwise in an ice bath. After the addition, the reaction was warmed to room temperature and stirred for 1 hour. After TLC detected the reaction was completed, the mixture was diluted with a small amount of water and extracted 3-4 times with EtOAc. The organic phases were combined, washed with brine, concentrated, and then purified by silica gel column chromatography to give a product (148 mg, yield: 73.8%). LC-MS (APCI): m/z=496.5 (M+1).sup.+.

Step 6: Synthesis of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-1-d-3-methylbutyl)boronic Acid (Compound I-2)

(22) The compound 17 (148 mg, 0.3 mmol) and isobutylboronic acid (79.63 mg, 0.78 mmol) were added to a reaction flask, and dissolved by adding 2 mL of methanol and 2 mL of n-hexane. 1N hydrochloric acid (0.4 mL, 0.4 mmol) was added, and the reaction was stirred at room temperature under nitrogen protection overnight. After TLC detected the reaction was completed, the methanol layer was separated, washed three times with n-heptane, concentrated to dryness under reduced pressure, which was dissolved by adding a small amount of 2N sodium hydroxide and washed three times with dichloromethane. The aqueous phase was adjusted to pH 2-3 with concentrated hydrochloric acid and extracted with dichloromethane for 3-4 times. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography. The product was dried and weighed 61 mg (yield: 56.3%). LC-MS (APCI): m/z=344.3 (M+1-H.sub.2O). .sup.1H NMR (400 MHz, DMSO) δ 8.94 (t, J=5.9 Hz, 1H), 8.69 (s, 1H), 7.65 (s, 1H), 7.55 (d, J=1.4 Hz, 2H), 4.02 (d, J=5.8 Hz, 2H), 1.62 (dt, J=13.3, 6.7 Hz, 1H), 1.35 (dd, J=13.0, 6.7 Hz, 1H), 1.25 (d, J=7.2 Hz, 1H), 0.86-0.79 (m, 6H).

Example 3 Preparation of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-2,2-d2-3-methylbutyl)boronic Acid (Compound I-3)

(23) ##STR00012##

(24) The specific synthetic steps are as follows:

(25) ##STR00013## ##STR00014##

Step 1: Synthesis of 1,1-d2-2-methylpropanol (Compound 19)

(26) Ethyl isobutyrate (580.8 mg, 5 mmol) was dissolved in anhydrous THF (20 mL), and LiAlD.sub.4 (230.9 mg, 5.5 mmol) was added portionwise in an ice bath. After the addition, the mixture was warmed to room temperature and reacted overnight. After TLC detected the reaction was completed, the reaction was quenched by adding sodium sulfate decahydrate under the ice bath. The insoluble material was removed by filtration, and the filtrate was concentrated to give a product (246 mg, yield: 64.73%), which was directly used in the next step.

Step 2: Synthesis of 1,1-d2-bromoisobutane (Compound 20)

(27) The compound 19 (850 mg, 11.17 mmol) and carbon tetrabromide (3.7 g, 11.17 mmol) were dissolved in dichloromethane (15 mL), and Triphenylphosphine (PPh.sub.3, 2.93 g, 11.17 mmol) was added portionwise in an ice bath. The reaction was stirred at low temperature for 1 hour under nitrogen protection, and then warmed to room temperature and reacted for 1 hour. After TLC detected the reaction was completed, a small amount of water was added to quench the reaction, and the aqueous phase was extracted three times with dichloromethane. The organic phases were combined, washed with brine, concentrated, and then purified by column chromatography to give a product (865 mg, yield: 56.2%).

Step 3: Synthesis of 1,1-d2-isobutylboronic Acid (Compound 21)

(28) The compound 20 (120 mg, 0.87 mmol) was dissolved in 5 mL of anhydrous THF, and magnesium turning (104.3 mg, 4.35 mmol) and a catalytic amount of iodine (1 granule) were added under nitrogen protection. The mixture was heated to 50° C. and stirred for 3 hours. After cooling to room temperature, the Grignard reagent was taken out with a syringe, and slowly added dropwise to trimethyl borate (180.8 mg, 1.74 mmol) in 5 mL of anhydrous THF at −15° C., and the reaction was stirred for 1 hour under nitrogen protection. After TLC detected the reaction was completed, the reaction was warmed to room temperature, and the pH was adjusted to 2-3 with dilute hydrochloric acid. The mixture was stirred for 15 minutes, and the organic phase was separated. The aqueous phase was extracted three times with ethyl acetate. The organic phases were combined, washed with brine and concentrated. The residue was purified by column chromatography to give a product (64 mg, yield: 70.7%).

Step 4: Synthesis of 1,1-d2-2-methylpropylboronic acid-(+)-pinanediol Ester (Compound 22)

(29) The compound 21 (110 mg, 1.06 mmol) and (1S, 2S, 3R, 5S)-(+)-2,3-pinanediol (179.8 mg, 1.06 mmol) were added to a reaction flask, and dissolved by adding 3 mL of n-heptane. The reaction was stirred at room temperature for 2 hours. After TLC detected the reaction was completed, the mixture was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and dried in vacuo to give a product (238 mg, yield: 94.2%).

Step 5: Synthesis of (S)-1-chloro-2,2-d2-3-methylbutylboronic Acid-(+)-pinanediol Ester (Compound 23)

(30) The compound 22 (238 mg, 1.0 mmol) was added to a reaction flask, and dissolved by adding 2 mL of anhydrous tetrahydrofuran and 0.5 mL of anhydrous dichloromethane. The temperature was lowered to −78° C. under nitrogen protection. A 2 M LDA solution (1.0 mL, 2.0 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 2 hours. A 1 M solution of zinc chloride in tetrahydrofuran (1.75 mL, 1.75 mmol) was slowly added dropwise. After the addition, the reaction was reacted for 2 hours at the low temperature. After TLC detected the reaction was completed, a small amount of 10% dilute sulfuric acid was added to quench the reaction, and the organic phase was separated. The aqueous phase was extracted with n-hexane. The organic phases were combined, washed with brine, concentrated, purified by silica gel column chromatography and dried in vacuo to give a product (265 mg, yield: 92.6%). LC-MS (APCI): m/z=287.7 (M+1).sup.+.

Step 6: Synthesis of (R)-1-(hexamethyldisilanyl)amino-2,2-d2-3-methylbutylboronic Acid Pinanediol Ester (Compound 24)

(31) The compound 23 (265 mg, 0.93 mmol) was added to a reaction flask, dissolved by adding 3 mL of anhydrous tetrahydrofuran, and lowered to −40° C. under nitrogen protection. Lithium bis(trimethylsilyl)amide (1.11 mL, 1.11 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 1 hour, and then warmed to room temperature and reacted for 1 hour. After TLC detected the reaction was completed, the mixture was filtered through a plug of silica gel, and the filtrate was evaporated to dryness to give a product (382.5 mg, yield: 100%), which was directly used in the next step.

Step 7: Synthesis of L-d2-leucine Boronic Acid-(+)-pinanediol Ester Trifluoroacetate Salt (Compound 25)

(32) The compound 24 (382.5 mg, 0.93 mmol) was added to a dry reaction flask, dissolved by adding 3 mL of diisopropyl ether, and lowered to −15° C. under nitrogen protection. Trifluoroacetic acid (424.1 mg, 3.72 mmol) was slowly added dropwise. After the addition, the reaction was reacted at this temperature for 2 hours and then warmed to room temperature and reacted overnight. After the filtration, the cake was washed with diisopropyl ether and dried in vacuo to give a white solid (202 mg, yield: 57.1%). LC-MS (APCI): m/z=268.2 (M+1).sup.+.

Step 8: Synthesis of 2,5-dichloro-N-[2-({(1R)-2,2-d2-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylene-1,3,2-benzodioxaborolan-2-yl]butyl}amino)-2-oxaethyl]benzamide (Compound 26)

(33) The compound 13 (100 mg, 0.402 mmol) and the compound 25 (152.4 mg, 0.402 mmol) were added to a reaction flask, TBTU (142 mg, 0.44 mmol) was added under nitrogen protection, and dissolved by adding 2 mL of anhydrous DMF. DIPEA (156 mg, 1.21 mmol) was added dropwise under an ice bath. After the addition, the reaction was warmed to room temperature and stirred for 1 hour. After TLC detected the reaction was completed, the mixture was diluted with a small amount of water and extracted 3-4 times with EtOAc. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography to give a product (183 mg, yield: 91.9%). LC-MS (APCI): m/z=497.4 (M+1).sup.+.

Step 9: Synthesis of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-2,2-d2-3-methylbutyl)boronic Acid (Compound I-3)

(34) The compound 26 (183 mg, 0.369 mmol) and isobutylboronic acid (97.9 mg, 0.96 mmol) were added to a reaction flask, and dissolved by adding 2 mL of methanol and 2 mL of n-hexane. 1N hydrochloric acid (0.5 ml, 0.5 mmol) was added, and the reaction was stirred at room temperature under nitrogen protection overnight. After TLC detected the reaction was completed, the methanol layer was separated, washed three times with n-heptane, concentrated to dryness under reduced pressure, which was dissolved by adding a small amount of 2N sodium hydroxide, and washed three times with dichloromethane. The aqueous phase was adjusted to pH 2-3 with concentrated hydrochloric acid and extracted with dichloromethane for 3-4 times. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography. The product was dried and weighed 75 mg (yield: 56.4%). LC-MS (APCI): m/z=345.2 (M+1-H.sub.2O). .sup.1H NMR (400 MHz, DMSO) δ 8.94 (t, J=5.9 Hz, 1H), 8.69 (s, 1H), 7.65 (s, 1H), 7.55 (d, J=1.4 Hz, 2H), 4.02 (d, J=5.8 Hz, 2H), 2.64 (s, 1H), 1.26 (m, 1H), 0.86-0.79 (m, 6H).

Example 4 Preparation of (R)-(1-(2-(2,5-dichlorobenzamido)-2,2-d2-acetamido)-1-d-3-methylbutyl)boronic Acid (Compound I-4)

(35) ##STR00015##

(36) The specific synthetic steps are as follows:

(37) ##STR00016##

Step 1: Synthesis of 2,5-dichloro-N-[2-({(1(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylene-1,3,2-benzodioxaborolan-2-yl]-1-d1-butyl}amino)-2-oxa-1,1-d2-ethyl]benzylamide (Compound 27)

(38) The compound 5 (100 mg, 0.402 mmol) and the compound 16 (154 mg, 0.402 mmol) were added to a reaction flask, TBTU (142 mg, 0.44 mmol) was added under nitrogen protection, and dissolved by adding 2 mL of anhydrous DMF. DIPEA (156 mg, 1.21 mmol) was added dropwise in an ice bath. After the addition, the reaction was warmed to room temperature and stirred for 1 hour. After TLC detected the reaction was completed, the mixture was diluted with a small amount of water and extracted 3-4 times with EtOAc. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography to give a product (199 mg, yield: 99%). LC-MS (APCI): m/z=498.2 (M+1).sup.+.

Step 2: Synthesis of (R)-(1-(2-(2,5-dichlorobenzamido)-2,2-d2-acetamido)-1-d-3-methylbutyl)boronic Acid (Compound I-4)

(39) The compound 27 (203 mg, 0.408 mmol) and isobutylboronic acid (108.4 mg, 1.06 mmol) were added to a reaction flask, and dissolved by adding 2 mL of methanol and 2 mL of n-hexane. 1N hydrochloric acid (0.5 mL, 0.5 mmol) was added, and the reaction was stirred at room temperature under nitrogen protection overnight. After TLC detected the reaction was completed, the methanol layer was separated, washed three times with n-heptane, concentrated to dryness under reduced pressure, which was dissolved by adding 2N sodium hydroxide, and washed three times with dichloromethane. The aqueous phase was adjusted to pH 2-3 with concentrated hydrochloric acid and extracted 3-4 times with dichloromethane. The organic phases were combined, washed with brine, concentrated, and then purified by silica gel column chromatography. The product was dried and weighed 96 mg (yield: 64.8%). LC-MS (APCI): m/z=346.4 (M+1-H.sub.2O). .sup.1H NMR (400 MHz, DMSO) δ 8.93 (s, 1H), 8.71 (d, J=11.4 Hz, 1H), 7.65 (s, 1H), 7.55 (d, J=1.3 Hz, 2H), 1.61 (dd, J=13.3, 6.6 Hz, 1H), 1.35 (dd, J=13.0, 7.0 Hz, 1H), 1.25 (d, J=7.1 Hz, 1H), 0.83 (dd, J=6.5, 1.9 Hz, 6H).

Example 5 Preparation of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-1-d-2,2-d2-3-methylbutyl)boronic Acid (Compound I-5)

(40) ##STR00017##

(41) The specific synthetic steps are as follows:

(42) ##STR00018##

Step 1: Synthesis of (S)-1-chloro-1-d1-2,2-d2-3-methylbutylboronic Acid-(+)-pinanediol Ester (Compound 28)

(43) The compound 22 (238 mg, 1.0 mmol) was added to a reaction flask, and dissolved by adding 2 mL of anhydrous tetrahydrofuran and 0.5 mL of deuterated dichloromethane. The temperature was lowered to −78° C. under nitrogen protection. A 2 M LDA solution (1.0 mL, 2.0 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 2 hours. A 1 M solution of zinc chloride in tetrahydrofuran (1.75 mL, 1.75 mmol) was slowly added dropwise. After the addition, the mixture was reacted at the low temperature for 2 hours. After TLC detected the reaction was completed, a small amount of 10% dilute sulfuric acid was added to quench the reaction, and the organic phase was separated. The aqueous phase was extracted with n-hexane. The organic phases were combined, washed with brine, concentrated, and then purified by silica gel column chromatography, and dried in vacuo to give a product (200 mg, yield: 69.7%). LC-MS (APCI): m/z=288.1 (M+1).sup.+.

Step 2: Synthesis of (R)-1-(hexamethyldisilanyl)amino-1-d1-2,2-d2-3-methylbutylboronic Acid Pinanediol Ester (Compound 29)

(44) The compound 28 (200 mg, 0.70 mmol) was added to a reaction flask, dissolved by adding 3 mL of anhydrous tetrahydrofuran, and lowered to −40° C. under nitrogen protection. Lithium bis(trimethylsilyl)amide (0.84 mL, 0.84 mmol) was slowly added dropwise. After the addition, the reaction was stirred for 1 hour, and then warmed to room temperature and reacted for 1 hour. After TLC detected the reaction was completed, the mixture was filtered through a plug of silica gel, and the filtrate was evaporated to dryness to give a product (288.6 mg, yield: 100%), which was directly used in the next step.

Step 3: Synthesis of L-d3-leucine Boronic Acid-(+)-pinanediol Ester Trifluoroacetate Salt (Compound 30)

(45) The compound 29 (288.6 mg, 0.7 mmol) was added to a dry reaction flask, dissolved by adding 3 mL of diisopropyl ether, and lowered to −15° C. under nitrogen protection.

(46) Trifluoroacetic acid (319.2 mg, 2.8 mmol) was slowly added dropwise. After the addition, the mixture was reacted at this temperature for 2 hours, and then warmed to room temperature and reacted overnight. After filtration, the filter cake was washed with diisopropyl ether and dried in vacuo to give a white solid (95 mg, yield: 35.6%). LC-MS (APCI): m/z=269.3 (M+1).sup.+.

Step 4: Synthesis of 2,5-dichloro-N-[2({(1R)-1-d1-2,2-d2-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylene-1,3,2-benzodioxaborolan-2-yl]butyl}amino)-2-oxaethyl]benzamide (Compound 31)

(47) The compound 13 (56 mg, 0.226 mmol) and the compound 30 (86.4 mg, 0.226 mmol) were added to a reaction flask, TBTU (80 mg, 0.25 mmol) was added under nitrogen protection, and dissolved by adding 2 mL of anhydrous DMF. DIPEA (87.6 mg, 0.34 mmol) was added dropwise in an ice bath. After the addition, the reaction was warmed to room temperature and stirred for 1 hour. After TLC detected the reaction was completed, the mixture was diluted with a small amount of water and extracted 3-4 times with EtOAc. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography to give a product (88 mg, yield: 78.6%). LC-MS (APCI): m/z=498.4 (M+1).sup.+.

Step 5: Synthesis of (R)-(1-(2-(2,5-dichlorobenzamido)acetamido)-1-d-2,2-d2-3-methylbutyl)boronic Acid (compound I-5)

(48) The compound 31 (88 mg, 0.177 mmol) and isobutylboronic acid (47 mg, 0.46 mmol) were added to a reaction flask, and dissolved by adding 1 mL of methanol and 1 mL of n-hexane. 1N hydrochloric acid (0.3 ml, 0.3 mmol) was added, and the reaction was stirred at room temperature under nitrogen protection overnight. After TLC detected the reaction was completed, the methanol layer was separated, washed three times with n-heptane, concentrated to dryness under reduced pressure, dissolved by adding a small amount of 2N sodium hydroxide, and washed three times with dichloromethane. The aqueous phase was adjusted to pH 2-3 with concentrated hydrochloric acid, and extracted 3-4 times with dichloromethane. The organic phases were combined, washed with brine, concentrated and then purified by silica gel column chromatography. The product was dried and weighed 32 mg (yield: 50.1%). LC-MS (APCI): m/z=346.4 (M+1-H.sub.2O). .sup.1H NMR (400 MHz, DMSO) δ 8.94 (t, J=5.9 Hz, 1H), 8.69 (s, 1H), 7.65 (s, 1H), 7.55 (d, J=1.4 Hz, 2H), 4.02 (d, J=5.8 Hz, 2H), 1.26 (m, 1H), 0.86-0.79 (m, 6H).

Example 6: Biological Activity Test

(1) 20S Proteasome Assay

(49) 25 μL of assay buffer containing human PA28 activator (Boston Biochem, final concentration 12 nM) and Ac-WLA-AMC ((35 selective substrate) (final concentration 15 μM) was added to 1 μL of test compounds dissolved in DMSO in a 384-well black microtiter plate at 37° C. Then, 25 μL of assay buffer containing human 20S proteasome (Boston Biochem, final concentration 0.25 nM) was added at 37° C. The assay buffer was consisted of 20 mM HEPES, 0.5 mM EDTA and 0.01% BSA with a pH of 7.4. The reaction was followed by a BMG Galaxy plate reader (37° C., 380 nm excitation, 460 nm emission, 20 gain). Percent inhibition was calculated based on the 0% inhibition (DMSO) and 100% inhibition (10 μM bortezomib) control groups.

(50) The proteasome inhibitory action of the compounds disclosed herein was tested according to the above assay. It was observed that the compounds disclosed herein exhibited an inhibitory activity against the proteasome. The compounds disclosed herein have inhibitory activity against the proteasome with an IC.sub.50 value of less than 50 nM.

(2) Metabolic Stability Evaluation

(51) Experiments in microsomes: Human liver microsomes: 0.5 mg/mL, Xenotech; Rat liver microsomes: 0.5 mg/mL, Xenotech; Coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; Magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

(52) Preparation of stock solution: A certain amount of powder of the example compounds was accurately weighed and dissolved in DMSO to 5 mM respectively.

(53) Preparation of phosphate buffer (100 mM, pH7.4): A pre-formulated 150 mL potassium dihydrogen phosphate (0.5 M) was mixed with 700 mL of dibasic potassium phosphate (0.5M). The pH of the mixture was adjusted to 7.4 with 0.5 M dibasic potassium phosphate solution. The mixture was diluted 5-fold with ultrapure water before use, and magnesium chloride was added to obtain the phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, pH 7.4.

(54) A NADPH regeneration system solution (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

(55) Preparation of stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, to which 812.5 μL human liver microsomes were added, respectively, and mixed to obtain a liver microsome dilution solution with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, to which 812.5 μL SD rat liver microsomes were added, respectively, and mixed to obtain a liver microsome dilution solution with a protein concentration of 0.625 mg/mL.

(56) Incubation of the samples: The stock solution of the respective compounds was respectively diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile, which were used as working solutions, ready for use. 398 μL of the dilution solution of human liver microsomes or rat liver microsomes were added to a 96-well incubation plate (N=2), respectively, and 2 μL of 0.25 mM working solution was added and mixed.

(57) Metabolic stability assay: 300 μL of pre-chilled stop solution was added to each well of a 96-well deep well plate and placed on ice as the stop plate. The 96 well incubation plate and NADPH regeneration system were placed in a 37° C. water bath box, shaken at 100 rpm and pre-incubated for 5 min. 80 μL of incubation solution was taken out from each well of the incubation plate and added to the stop plate, mixed, and replenished with 20 μL of NADPH regeneration system solution as a 0-min sample. 80 μL of NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and counting was started. The corresponding compounds had a reaction concentration of 1 μM and the protein concentration was 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min after reaction, respectively, added to the stop plate, and vortexed for 3 minutes to terminate the reaction. The stop plate was centrifuged for 4 min at 5000× g at 4° C. 100 μL of the supernatant was added to a 96-well plate to which 100 μL of distilled water was previously added, mixed, and analyzed by LC-MS/MS.

(58) Data analysis: The peak areas of the corresponding compounds and internal standard were detected by LC-MS/MS system, and the ratio of the peak area of the compounds to the internal standard was calculated. The slope was measured by plotting the natural logarithm of the percent of remained compound versus time, and t.sub.1/2 and CL.sub.int were calculated according to the formula below, where V/M is equal to 1/protein concentration.

(59) $t_{1/2}=-\frac{0.693}{\mathrm{Peak}\mathrm{area}},{\mathrm{CL}}_{\mathrm{int}}=\frac{0.693}{t_{1/2}}.Math.\frac{V}{M}.$

(60) The metabolic stability in human and rat liver microsomes was evaluated by simultaneously testing and comparing the compounds disclosed herein and the non-deuterated compound. The half-life and liver intrinsic clearance as indicators of metabolic stability are shown in the table. The non-deuterated compound Ixazomib was used as a control in the table. As shown in the table, the compounds disclosed herein can significantly improve the metabolic stability by comparison with Ixazomib in human and rat liver microsome experiments.

(61) TABLE-US-00001 TABLE Comparison Table of Indicators for Metabolic Stability of Compounds of Examples 1-4 Experiment in Experiment in human liver microsomes rat liver microsomes Number t.sub.1/2 CL.sub.int t.sub.1/2 CL.sub.int Ixazomib 28.1 49.4 14.3 97.3 I-1 — — 17.1 81.3 I-2 29.8 46.5 15.3 90.5 I-3 29.3 47.3 — — I-4 28.7 48.3 15.4 90.0 I-5 29.3 47.3 14.2 97.8

(3) Pharmacokinetic Experiment in Rats

(62) Experimental objective: After administration of Ixazomib or Example compounds to rats, the pharmacokinetic behavior of the compounds disclosed herein was investigated.

(63) Experimental Animals:

(64) Species and strains: SD rat, grade: SPF grade

(65) Gender and quantity: male, 6

(66) Weight range: 180 to 220 g (the actual weight range was from 187 to 197 g)

(67) Source: shanghai sippr bk laboratory animals ltd.

(68) Laboratory and animal certificate number: SCXK (Shanghai) 2013-0016.

(69) Experiment procedure:

(70) Before blood samples were collected, 20 μL of 2 M sodium fluoride solution (esterase inhibitor) was previously added to an EDTA-K2 anticoagulant tube, dried in an 80° C. oven, and placed in a 4° C. refrigerator.

(71) Rats (male, weighted 187 to 197 g) were randomly divided into 2 groups, and were fasted overnight in the afternoon before the experiment, but were allowed to drink water freely. Food was given 4 hours after the administration. Group A was given 3 mg/kg of Ixazomib, and group B was given 3 mg/kg of the Example compounds. About 100-200 μL of blood was taken from the orbital vein of rats at 15 min, 30 min, and 1, 2, 3, 5, 8 and 10 h after administration, placed in a 0.5 mL Eppendorf tube with EDTA-K2 anticoagulant and mixed immediately. After anticoagulation, the tube was gently inverted 5-6 times as quickly as possible. After the blood was taken, it was placed in an ice box, and then within 30 min, the blood sample was centrifuged for 10 min at 4000 rpm and at 4° C. to separate the plasma. Immediately after collection of all plasma, it was stored at −20° C. The concentration of the drug in plasma at each time point was determined after sample collection at all time points.

(72) Based on the data obtained as described above (the average concentration of the drug in plasma after administration versus time), pharmacokinetics-related parameters of male SD rats after the i.g. administration of Ixazomib (3 mg/kg) and the Example compounds (3 mg/kg) were calculated using the Winnonin software according to non-compartment statistical moment theory.

(73) The experiments showed that, as compared with Ixazomib, the compounds disclosed herein have excellent pharmacokinetic properties, and thus are more suitable as compounds for inhibiting proteasomes, and are further suitable for the preparation of a medicament for treating a proteasome-mediated disease.

(74) It should be understood that these examples are only for illustrating the present disclosure and are not intended to limit the scope disclosed herein. Experimental methods that do not specify specific conditions in the examples are generally based on conventional conditions or conditions recommended by the manufacturer. Parts and percentages are parts by weight and percentages by weight unless otherwise indicated.

(75) The above content is a further detailed description disclosed herein in combination with specific embodiments, and it cannot be assumed that the specific implementation disclosed herein is limited to these descriptions. For a person of ordinary skill in the art to which the present disclosure belongs, a number of simple deductions or substitutions can be made without departing from the concept disclosed herein, and should all be considered as falling within the protection scope disclosed herein.