Tripeptide Propylene Oxide Derivative and Preparation Method and Application thereof

Abstract

The present invention discloses a tripeptide propylene oxide derivative or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof. The structure of the tripeptide propylene oxide derivative is shown in formula I. Compared with the prior art, the present invention provides tripeptide epoxy ketone compounds with a novel structure and a function of inhibiting proteasome. As 20S proteasome inhibitors, the tripeptide epoxy ketone compounds can block tumor cell proliferation and induce tumor cell apoptosis, so the tripeptide epoxy ketone compounds can be used for the treatment and prevention of a plurality of human and animal diseases such as malignant tumors, and the effect is significantly better.

Claims

1. A tripeptide propylene oxide derivative or a pharmaceutically acceptable salt thereof, wherein the structure of the tripeptide propylene oxide derivative is shown in formula I, ##STR00057## where: R.sub.1 is selected from a group consisting of hydrogen, deuterium, C.sub.1-10 alkyl, C.sub.3-6 cycloalkyl, heterocycloalkyl, heterocyclyl, aryl or benzyl, and the C.sub.1-10 alkyl, C.sub.3-6 cycloalkyl, heterocycloalkyl, heterocyclyl, aryl or benzyl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, C.sub.1-4 alkylthio, cyano, nitro, hydroxyl, sulfydryl, amino and halogen; R.sub.2 is selected from a group consisting of hydrogen, deuterium or C1-10 heteroalkyl, and the C.sub.1-10 heteroalkyl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, C.sub.1-4 alkylthio, cyano, nitro, hydroxyl, sulfydryl, amino and halogen; R.sub.3 is selected from a group consisting of hydrogen, deuterium or C.sub.1-10 heteroalkyl, and the C.sub.1-10 heteroalkyl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, C.sub.1-4 alkylthio, cyano, nitro, hydroxyl, sulfydryl, amino and halogen; Z is selected from one of the following fragments: ##STR00058## and P is selected from a group consisting of hydrogen, C.sub.1-10 alkyl, C.sub.1-10 alkoxyl, aryl or heteroaryl; and the C.sub.1-10 alkyl, C.sub.1-10 alkoxyl, aryl or heteroaryl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, halogen and halogenated C.sub.1-4 alkyl.

2. The tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1, wherein R.sub.1 is selected from a group consisting of hydrogen, C.sub.1-10 alkyl, phenyl, naphthyl, indolyl, thiazolyl, thienyl, benzothienyl, imidazolyl or benzyl, and the C.sub.1-10 alkyl, phenyl, naphthyl, indolyl, thiazolyl, thienyl, benzothienyl, imidazolyl or benzyl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, cyano, nitro, hydroxyl, sulfydryl, amino and halogen; R.sub.2 is selected from a group consisting of hydrogen or C.sub.1-10 heteroalkyl, and the C.sub.1-10 heteroalkyl is optionally substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl and C.sub.1-4 alkylthio; R.sub.3 is hydrogen or C.sub.1-10 heteroalkyl, and the C.sub.1-10 heteroalkyl is substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl or C.sub.1-4 alkylthio; and P is selected from a group consisting of hydrogen, morpholinyl, methylisoxazolyl, 2-methylthiazolyl, 2,5-dichlorophenyl and pyrazinyl.

3. The tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1, wherein R.sub.1 is selected from a group consisting of hydrogen, C.sub.1-4 alkyl, phenyl, indolyl, thiazolyl, thienyl, benzothienyl, imidazolyl or benzyl, and the C.sub.1-4 alkyl, phenyl, indolyl, thiazolyl, thienyl, benzothienyl, imidazolyl and benzyl that is substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl, nitro or halogen; R.sub.2 is hydrogen or C.sub.1-4 heteroalkyl, and the C.sub.1-4 heteroalkyl is substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl or C.sub.1-4 alkylthio; and R.sub.3 is hydrogen or C.sub.1-4 heteroalkyl, and the C.sub.1-4 heteroalkyl is substituted or unsubstituted by C.sub.1-4 alkyl, C.sub.1-4 alkoxyl or C.sub.1-4 alkylthio.

4. The tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1, wherein the tripeptide propylene oxide derivative is selected from a group consisting of ##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063##

5. The tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1, wherein the tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof is synthesized according to the following route: ##STR00064## ##STR00065## where the definitions of the groups P, R.sub.1, R.sub.2, R.sub.3 and Z are as described in claim 1.

6. The tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1, wherein the tripeptide propylene oxide derivative is used for a proteasome inhibiting drug.

7. A pharmaceutical composition, comprising the tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof claim 1 and a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, wherein the composition is used for treating immune-related diseases.

9. The pharmaceutical composition according to claim 7, wherein the composition further comprises antigen peptides; the antigen peptides are produced by proteasome in vivo.

10. A method for treating a disease comprising a step of administrating effective amount of the tripeptide propylene oxide derivative or pharmaceutically acceptable salt thereof of claim 1 to a subject in need thereof; wherein the disease is selected from a group consisting of inflammation, cancer and hyperproliferative disease.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0107] FIG. 1 Results of the pharmacodynamics of the compound of the present invention in ICR mice.

[0108] FIG. 2 Results of the drug effect of the compound of the present invention in nude mice.

[0109] FIG. 3 Results of changes in body weight after continuous administration of the compound of the present invention to nude mice.

DETAILED DESCRIPTION OF THE INVENTION

[0110] The technical solutions of the present invention are further described below through specific implementations.

PART I SYNTHESIS OF COMPOUNDS

[0111] The compounds of the present invention can be prepared according to the following process:

I. Preparation of the Compound (II)

[0112] ##STR00010##

[0113] 1. Preparation of the Compound (II-3):

[0114] The compound (II-1) and HOBt are dissolved in anhydrous DCM and the reaction solution is stirred at −5° C. for 10 min. EDI.HCl is added at the temperature and the reaction solution is stirred for 15-20 min. The compound (II-2) is added and the reaction solution is stirred for 15-20 min. DIPEA is added and the reaction solution is stirred for 20 min. The reaction solution is moved to room temperature for reaction. After the reaction is complete, the reaction solution is poured into water and extracted with DCM. The organic phases are combined and washed with dilute HCl, a NaHCO.sub.3 solution and saturated brine respectively, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound (II-3).

[0115] 2. Preparation of the Compound (II-4):

[0116] The compound (II-3) is dissolved in anhydrous DCM.TFA is added dropwise slowly at −5° C. After stirring for 0.5 h, the temperature is raised to room temperature and the reaction solution is stirred for 3 h and then tested. After the reaction is complete, the reaction solution is concentrated to obtain brown-red oil. The brown-red oil is slowly added to methyl tert-butyl ether, and the reaction solution is vigorously stirred to obtain a white solid. The white solid is filtered to obtain the compound (II-4).

[0117] 3. Preparation of the Compound (II-6):

[0118] P group substituted carboxylic acid, i.e. the compound (II-5) and HOBt are dissolved in anhydrous DCM and the reaction solution is stirred at −5° C. for 10 min. EDI.HCl is added at the temperature and the reaction solution is stirred for 15-20 min. The compound (II-4) is added and the reaction solution is stirred for 15-20 min. DIPEA is added and the reaction solution is stirred for 20 min. The reaction solution is moved to room temperature for reaction. After the reaction is complete, the reaction solution is poured into water and extracted with DCM. The organic phases are combined and washed with dilute HCl, a NaHCO.sub.3 solution and saturated brine respectively, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound (II-6).

[0119] 4. Preparation of the Compound (II):

[0120] The compound (II-6) is dissolved in MeOH/H.sub.2O. A LiOH aqueous solution is added dropwise at 0° C. and the reaction solution is stirred for 2 h. The temperature is raised to room temperature for reaction for a certain time. Water is added and the pH is adjusted to 6-7 with hydrochloric acid. The reaction solution is extracted with ethyl acetate. The organic phase is washed with saturated brine, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound (II).

II. Preparation of the Compound (III)

[0121] ##STR00011##

[0122] 1. Preparation of the Compound (III-2):

[0123] The compound (III-1) and HOBt are dissolved in DCM. EDC.HCl is added. After stirring at −5° C. for 15 min, dimethylhydroxylamine hydrochloride is added. DIPEA is added after 15 min. Reaction is performed at low temperature for 25 min. After the reaction is complete at room temperature, the reaction solution is extracted with DCM. The organic phase is washed with IN HCl, 5% NaHCO.sub.3 and saturated brine, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound (III-2).

[0124] 2. Preparation of the Compound (III-3):

[0125] The compound (III-2) is dissolved with tetrahydrofuran. At −20° C., ethyl magnesium bromide is added dropwise. After dropwise adding, the temperature is raised to room temperature for reaction. After the reaction is complete, IN HCl is slowly added dropwise to quench the reaction. Extraction is performed with ethyl acetate. The extract is washed with saturated brine. The organic phase is dried and concentrated to obtain the compound (III-3).

[0126] 3. Preparation of the Compound (III-4):

[0127] The compound (III-3) is dissolved with tetrahydrofuran. Piperidine acetate and piperidine are added. Paraformaldehyde is added in batches. After refluxing for 3 h, an appropriate amount of water is added and extraction is performed with ethyl acetate. The extract is washed with IN HCl and saturated brine respectively. The organic phase is dried and concentrated to obtain the compound (III-4).

[0128] 4. Preparation of the Compound (III-5):

[0129] The compound (III-4) is dissolved in toluene. Aluminum isopropoxide and isopropanol are added. The reaction is performed at 50° C. After the reaction is complete, extraction is performed with water and ethyl acetate. The extract is washed with IN HCl and saturated brine. The organic phase is dried and concentrated to obtain the compound (III-5).

[0130] 5. Preparation of the Compound (III-6):

[0131] The compound (III-5) is dissolved in DCM. Then vanadium acetylacetonate is added. Under nitrogen protection, the reaction solution is cooled to 0° C. in an ice bath. Tert-Butyl hydroperoxide is slowly added dropwise. After the reaction is complete, an appropriate amount of water is added and extraction is performed with dichloromethane. The extract is washed with saturated sodium thiosulfate and saturated brine respectively. The organic phase is dried, concentrated and purified to obtain the compound (II-6).

[0132] 6. Preparation of the Compound (III-7):

[0133] The compound (III-6) is dissolved in dimethyl sulfoxide. Diisopropylethylamine is added. Pyridine sulfur trioxide is added in batches in an ice bath. The temperature is raised to room temperature for reaction. After the reaction is complete, an appropriate amount of water is added and extraction is performed with ethyl acetate. The extract is washed with IN HCl and saturated brine. The organic phase is dried and concentrated to obtain the compound (III-7).

[0134] 7. Preparation of the Compound (III):

[0135] The compound (II-7) is dissolved in anhydrous DCM. TFA is slowly added dropwise at −5° C. After stirring for 0.5 h, the temperature is raised to room temperature and the reaction solution is stirred for 3 h and then tested. After the reaction is complete, the reaction solution is concentrated to obtain brown-red oil. The brown-red oil is slowly added to methyl tert-butyl ether, and the reaction solution is vigorously stirred to obtain a white solid. The white solid is filtered to obtain the compound (III).

III. Preparation of the Compound (I)

[0136] ##STR00012##

[0137] Preparation of the Compound (I):

[0138] The compound (II) and HOBt are dissolved in anhydrous DCM and the reaction solution is stirred at −5° C. for 10 min. EDI.HCl is added at the temperature and the reaction solution is stirred for 15-20 min. The compound (III) is added and the reaction solution is stirred for 15-20 min. DIPEA is added and the reaction solution is stirred for 20 min. The reaction solution is moved to room temperature for reaction. After the reaction is complete, the reaction solution is poured into water and extracted with DCM. The organic phases are combined and washed with dilute HCl, a NaHCO.sub.3 solution and saturated brine respectively, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound (I).

[0139] The preparation process of the compound of the present invention is described with the synthesis of specific compounds as follows:

I. Preparation of Acid Fragments

[0140] The preparation of N-2,5-dichlorophenyl-2-formyl-O-methyl-L-serine-S-methyl-L-cysteine is taken as an example:

##STR00013##

[0141] The compound 1 (1 g, 4.56 mmol) and HOBt (0.92 g, 6.84 mmol) are dissolved in anhydrous DCM (50 mL) and the reaction solution is stirred at −5° C. for 10 min. EDI.HCl (1.31 g, 6.84 mmol) is added at the temperature and the reaction solution is stirred for 15-20 min. The compound 2 (0.85 g, 4.56 mmol) is added and the reaction solution is stirred for 15-20 min. DIPEA (2.26 mL, 13.68 mmol) is added and the reaction solution is stirred for 20 min. The reaction solution is moved to room temperature for reaction. After the reaction is complete, the reaction solution is poured into ice water and extracted with DCM. The organic phases are combined and washed with 0.4 N HCl, 5% NaHCO.sub.3 and saturated brine respectively, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound 3.

[0142] The compound 3 (1 g, 2.85 mmol) is dissolved in anhydrous DCM (30 mL). TFA (5.13 mL) is added dropwise slowly at −5° C. After stirring for 0.5 h, the temperature is raised to room temperature and the reaction solution is stirred for 3 h. After the reaction is complete, the reaction solution is concentrated to obtain brown-red oil. The brown-red oil is slowly added to methyl tert-butyl ether, and the reaction solution is vigorously stirred to obtain a white solid. The white solid is filtered to obtain the compound 4.

[0143] The compound 5 (1 g, 5.24 mmol) and HOBt (1.06 g, 7.86 mmol) are dissolved in anhydrous DCM (50 mL) and the reaction solution is stirred at −5° C. for 10 min. EDI.HCl (1.50 g, 7.86 mmol) is added at the temperature and the reaction solution is stirred for 15-20 min. The compound 4 (1.91 g, 5.24 mmol) is added and the reaction solution is stirred for 15-20 min. DIPEA (2.47 mL, 15.72 mmol) is added and the reaction solution is stirred for 20 min. The reaction solution is moved to room temperature for reaction. After the reaction is complete, the reaction solution is poured into ice water and extracted with DCM. The organic phases are combined and washed with 0.4 N HCl, 5% NaHCO.sub.3 and saturated brine respectively, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound 6.

[0144] The compound 6 (1 g, 2.36 mmol) is dissolved in MeOH/H.sub.2O (20 mL/5 mL). A LiOH.H.sub.2O (0.14 g, 3.31 mmol) in H.sub.2O (1 mL) solution is added dropwise at 0° C. and the reaction solution is stirred for 2 h. The temperature is raised to room temperature for reaction for a certain time. Water is added and the pH is adjusted to 6-7 with hydrochloric acid. The reaction solution is extracted with ethyl acetate. The organic phase is washed with saturated brine, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound 7, yield 90%, m.p.: 42.3-43.7° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.13 (s, 3H), 3.02 (d, J=4.8 Hz, 2H), 3.43 (s, 3H), 3.61 (dd, J=17.2, 9.9 Hz, 2H), 3.93 (d, J=9.0 Hz, 2H), 4.89-4.78 (m, 2H), 7.35 (s, 2H), 7.41 (d, J=4.5 Hz, 1H), 7.44 (d, J=8.7 Hz, 1H), 7.58 (d, J=7.5 Hz, 1H), 7.64 (d, J=4.2 Hz, 1H); MS (ESI) m/z: 410.2 [M+H].sup.+.

[0145] The synthetic methods of all acid fragment compounds in the present invention are similar to that of 7.

[0146] The specific compounds synthesized and names thereof are as follows.

TABLE-US-00001 No. Structure Chemical name and analysis data 8 [00014] N-5-methylisoxazolyl-3-formyl-O-methyl- L-seryl-S-methyl-L-cysteine Yield 90%, m.p.: 50.2-51.8° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.13 (s, 3H), 2.49 (s, J = 4.9 Hz, 3H), 3.07 − 2.91 (m, 2H), 3.40 (s, 3H), 3.67 − 3.56 (m, J = 8.8, 6.7 Hz, 1H), 3.88 (dd, J = 9.3, 4.2 Hz, 1H), 4.94 − 4.77 (m, 2H), 6.44 (s, 1H), 7.38 (s, J = 7.3, 3.5 Hz, 1H),7.50 (s, J = 11.4 Hz, 1H), 7.76 (s, J = 15.7, 7.5 Hz, 1H); MS (ESI) m/z: 346.1 [M + H].sup.+. 9 [00015] N-pyrazinyl-2-formyl-O-methyl-L-seryl-S- methyl-L-cysteine Yield 90%, m.p.: 80.5-82.1° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.04 (s, 3H), 3.15 − 2.95 (m, 2H), 3.43 (s, 3H), 3.72 − 3.58 (m, 1H), 3.96 − 3.89 (m, J = 9.4, 7.4, 4.4 Hz, 1H), 4.98 − 4.81 (m, 2H), 7.43 (s, 1H), 7.55 (s, J = 7.5 Hz, 1H), 8.60 (s, 1H), 8.69 − 8.59 (m, 1H), 8.78 (s, J = 1.6 Hz, 1H), 9.37 (s, J = 4.0 Hz, 1H); MS (ESI) m/z: 343.1 [M + H].sup.+. 10 [00016] N-2,5-dichlorophenyl-2-formyl-S-methyl- L-cysteinyl-O-methyl-L-serine Yield 90%, m.p.: 42.3-43.7° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.10 (dd, J = 14.3, 2.3 Hz, 3H), 2.49 − 2.41 (m, 3H), 2.96 − 2.77 (m, 2H), 3.24 (s, 1H), 3.53 (dd, J = 8.4, 3.8 Hz, 1H), 3.67 (dd, J = 9.4, 4.3 Hz, 1H), 4.43 (dt, J = 8.3, 4.5 Hz, 0H), 4.85 − 4.69 (m, 1H), 6.67 − 6.53 (m, 1H), 8.71 − 8.36 (m, 2H); MS (ESI) m/z: 410.2 [M + H].sup.+. 11 [00017] N-pyrazinyl-2-formyl-S-methyl-L- cysteinyl-O-methyl-L-serine Yield 90%, m.p.: 80.5-82.1° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.04 (d, J = 30.8 Hz, 3H), 2.93 (2, 2H), 3.25 (d, J = 2.0 Hz, 3H), 3.52 (dd, J = 6.7, 2.7 Hz, 1H), 3.71 − 3.66 (m, 1H), 4.46 (d, J = 3.6 Hz, 1H), 4.95 − 4.81 (m, 1H), 8.64 (dt, J = 27.7, 13.8 Hz, 1H), 8.84 − 8.74 (m, 2H), 8.91 (d, J = 2.3 Hz, 1H), 9.21 (s, 1H), 12.57 (s, 1H); MS (ESI) m/z: 343.1 [M + H].sup.+. 12 [00018] N-5-methylisoxazolyl-3-formyl-S-methyl- L-cysteinyl-O-methyl-L-serine Yield 90%, m.p.: 50.2-51.8° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.10 (dd, J = 14.3, 2.3 Hz, 3H), 2.49 − 2.41 (m, 3H), 2.96 − 2.77 (m, 2H), 3.24 (s, 1H), 3.53 (dd, J = 8.4, 3.8 Hz, 1H), 3.67 (dd, J = 9.4, 4.3 Hz, 1H), 4.43 (dt, J = 8.3, 4.5 Hz, 0H), 4.85 − 4.69 (m, 1H), 6.67 − 6.53 (m, 1H), 8.71 − 8.36 (m, 2H); MS (ESI) m/z: 346.1 [M + H].sup.+. 13 [00019] N-2-methylithiazolyl-5-formyl-S-methyl-L- cysteinyl-S-methyl-L-cysteine Yield 90%, m.p.: 55.3-56.4° C.; .sup.1H NMR (400 MHz, DMSO) δ 1.99 (s, J = 3.6, 1.8 Hz, 3H), 2.04 (s, 3H), 2.60 (s, 3H), 2.85 − 2.75 (m, 2H), 2.95 − 2.84 (m, 2H), 4.62 − 4.55 (m, 1H), 4.74 − 4.63 (m, 1H), 7.29 (s, J = 0.8 Hz, 1H), 7.58 (d, J = 7.5 Hz, 1H), 7.88 (d, J = 10.0 Hz, 1H), 8.08 (s, 1H); MS (ESI) m/z: 378.1 [M + H].sup.+. 14 [00020] N-2,5-dichlorophenyl-2-formyl-S-methyl- L-cysteinyl-S-methyl-L-cysteine Yield 90%, m.p.: 44.3-45.7° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.08 (s, 1H), 2.10 (s, 2H), 2.12 (s, 3H), 2.85 − 2.63 (m, 2H), 2.97 − 2.86 (m, 2H), 4.54 − 4.39 (m, J = 12.8, 7.9, 5.1 Hz, 1H), 4.80 − 4.65 (m, 1H), 7.51 (dt, J =4.0, 1.5 Hz, 1H), 7.55 (t, J = 1.4 Hz, 2H), 8.43 (d, J = 7.8 Hz, 1H), 8.52 (d, J = 8.2 Hz, 1H), 8.84 (d, J = 8.6 Hz, 1H); MS (ESI) m/z: 426.1 [M + H].sup.+. 15 [00021] N-pyrazinyl-2-formyl-S-methyl-L- cysteinyl-S-methyl-L-cysteine Yield 90%, m.p.: 55.5-56.3° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.08 (dd, J = 7.2, 2.5 Hz, 6H), 2.81 − 2.68 (m, 1H), 3.01 − 2.83 (m, 3H), 4.50 − 4.40 (m, 1H), 4.92 − 4.77 (m, 1H), 8.63 (d, J = 7.9 Hz, 1H), 8.77 (d, J = 6.7 Hz, 1H), 8.82 (d, J = 8.9 Hz, 1H), 8.92 (d, J = 2.4 Hz, 1H), 9.21 (s, 1H); MS (ESI) m/z: 359.1 [M + H].sup.+. 16 [00022] N-5-methylisoxazolyl-3-formyl-S-methyl- L-cysteinyl-S-methyl-L-cysteine Yield 90%, m.p.: 59.-60.2° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.07 (dd, J = 6.2, 2.3 Hz, 6H), 2.48 (s, J = 8.3 Hz, 3H), 2.82 − 2.71 (m, 1H), 2.96 − 2.82 (m, J = 13.5, 8.2, 4.2 Hz, 3H), 4.51 − 4.36 (m, 1H), 4.80 − 4.65 (m, 1H), 6.58 (s, J = 0.7 Hz, 1H), 8.50 (s, J = 7.8 Hz, 1H), 8.59 (s, J = 15.3, 5.4 Hz, 1H), 12.95 (s, 1H); MS (ESI) m/z: 362.1 [M + H].sup.+. 17 [00023] N-5-methylisoxazolyl-3-formyl-O-methyl- L-seryl-glycine Yield 90%, m.p.: 150-15l° C.; .sup.1H NMR (400 MHz, DMSO) 82.46 (d, J = 0.7 Hz, 3H), 3.26 (s, 3H), 3.52 (dd, J = 9.8, 3.9 Hz, 1H), 3.66 (dd, J = 9.8, 5.2 Hz, 1H), 4.14 (dd, J = 6.0, 3.6 Hz, 2H), 4.49 − 4.40 (m, 1H), 6.54 (s, J = 0.9 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.72 (t, J = 6.0 Hz, 1H); MS (ESI) m/z: 286.1 [M + H].sup.+. 18 [00024] N-5-methylisoxazolyl-3-formyl-glycyl-O- methyl-L-serine Yield 90%, m.p.: 149.1-151.7° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.46 (d, J = 0.7 Hz, 3H), 3.26 (s, 3H), 3.52 (dd, J = 9.8, 3.9 Hz, 1H), 3.66 (dd, J = 9.8, 5.2 Hz, 1H), 3.92 (dd, J = 6.0, 3.6 Hz, 2H), 4.49 − 4.40 (m, 1H), 6.54 (s, J = 0.9 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 8.72 (t, J = 6.0 Hz, 1H); MS (ESI) m/z: 595.5 [M + H].sup.+. 19 [00025] N-5-methylisoxazolyl-3-formyl-S-methyl- L-cysteinyl-glycine Yield 90%, m.p.: 150.1-151.4° C.; .sup.1H NMR (400 MHz, DMSO) δ 2.08 (s, 2H), 2.48 (s, 3H), 2.98 − 2.80 (m, J = 23.6, 13.8, 7.1 Hz, 2H), 3.78 (dd, J = 5.7, 3.4 Hz, 2H), 4.69 (td, J = 9.5, 4.4 Hz, 1H), 6.59 (s, J = 0.8 Hz, 1H), 8.49 (t, 1H), 8.65 (d, J = 8.6 Hz, 1H), 12.60 (s, 1H); MS (ESI) m/z: 302.1 [M + H].sup.+. 20 [00026] N-2-methylthiazolyl-5-formyl−S-methyl-L- cysteinyl-glycine Yield 90%, m.p.: 129.4-131.6° C.; .sup.1H NMR (400 MHz, CD.sub.3OD_SPE) 82.14 (s, J = 7.8 Hz, 3H), 2.72 (s, 3H), 2.91 − 2.80 (m, J = 14.0, 7.3 Hz, 1H), 3.14-3.00 (m, 1H), 3.96 (d, J = 13.6, 5.5 Hz, 2H), 4.81 − 4.72 (m, J = 9.7, 4.9 Hz, 1H), 8.23 (s, 1H); MS (ESI) m/z: 318.1 [M + H].sup.+.

II. Preparation of Amine Fragments

[0147] The preparation of compound (S)-2-amino-4-methyl-1-((R)-2-methyloxirane-2-yl)pentan-1-one 2,2,2-trifluoroacetate (28) is taken as an example:

##STR00027##

[0148] The compound 21 (10.0 g, 43.23 mmol) and HOBt (5.8 g, 43.23 mmol) are dissolved in DCM (100 mL). EDC.HCl (11.65 g, 64.85 mmol) is added. After stirring at −5° C. for 15 min, dimethylhydroxylamine hydrochloride (4.21 g, 43.23 mmol) is added. DIPEA (13.97 g, 108.08 mmol) is added after 15 min. Reaction is performed at low temperature for 25 min. After the reaction is complete at room temperature, the reaction solution is extracted with DCM. The organic phase is washed with 1N HCl, 5% NaHCO.sub.3 and saturated brine, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound 22.

[0149] N-tert-butoxycarbonyl-L-leucine-N′-methoxyl-N′-formamide 22 (0.5 mol) is weighed, added into a reaction flask and dissolved with 500 mL of tetrahydrofuran. At −20° C., ethyl magnesium bromide (2.0 M, 750 mL) is added dropwise. After dropwise adding, the temperature is raised to room temperature for reaction overnight. 1N HCl is added dropwise slowly to quench the reaction. Extraction is performed with ethyl acetate. The extract is washed with saturated brine. The organic phase is dried and concentrated to obtain the compound 23.

[0150] The compound 23 (0.4 mol) is weighed and dissolved with 400 mL tetrahydrofuran. Piperidine acetate (1.5 mol), piperidine (1.0 mol) and paraformaldehyde (2.0 mol) are added. After refluxing for 3 h, paraformaldehyde (2.0 mol) is added. The reaction is tested by TLC to completion. An appropriate amount of water is added and extraction is performed with ethyl acetate. The extract is washed with 1N HCl and saturated brine 1 time respectively. The organic phase is dried and concentrated to obtain the compound 24.

[0151] Aluminum isopropoxide (0.3 mol) and isopropanol (3 mol) are weighed, added to 200 mL of toluene and the compound 24 (0.3 mol), dissolved with 100 mL of toluene, and added dropwise to the reaction system at room temperature. After dropwise adding, the reaction is performed at 50° C., and tested by TLC to completion. An appropriate amount of water is added and then extraction is performed with ethyl acetate. The extract is washed with 1N HCl and saturated brine 1 time respectively. The organic phase is dried and concentrated to obtain the compound 25.

[0152] The compound 25 (0.2 mol) is weighed and dissolved in 200 mL of dichloromethane. Then vanadium acetylacetonate (0.04 mol) is added. Under nitrogen protection, the reaction solution is cooled to 0° C. in an ice bath. Tert-Butyl hydroperoxide is added dropwise slowly. The reaction solution is stirred more vigorously overnight, and tested by TLC that the raw materials disappear. An appropriate amount of water is added and extraction is performed with dichloromethane. The extract is washed with saturated sodium thiosulfate and saturated brine respectively. The organic phase is dried, concentrated and purified to obtain the compound 26.

[0153] The compound 26 (0.12 mol) is dissolved in 100 mL of dimethyl sulfoxide. Diisopropylethylamine (0.24 mol) is added. Pyridine sulfur trioxide (0.24 mol) is added in batches in an ice bath. The temperature is raised to room temperature for reaction. After the reaction is complete tested by TLC, an appropriate amount of water is added and extraction is performed with ethyl acetate. The extract is washed with IN HCl and saturated brine. The organic phase is dried and concentrated to obtain the compound 27.

[0154] The compound 27 (1.0 g, 3.69 mmol) is dissolved in anhydrous DCM (10 mL). TFA (3 mL) is added dropwise slowly at −5° C. After stirring for 0.5 h, the temperature is raised to room temperature and the reaction solution is stirred for 3 h and then tested. After the reaction is complete, the reaction solution is concentrated to obtain brown-red oil. The brown-red oil is slowly added to methyl tert-butyl ether, and the reaction solution is vigorously stirred to obtain a white solid. The white solid is filtered to obtain the compound 28. The yield is 85%, m.p.: 83-84° C. .sup.1H NMR (400 MHz, CDCl3): δ 0.97 (—CH3, d, J=6.4 Hz, 3H), 1.28 (—CH3, d, J=14.1 Hz, 3H), 1.83-1.64 (—CH, m, 1H), 1.92-1.84 (—CH2, m, 2H), 2.93 ((—CH, d, J=4.5 Hz, 1H) , 3.16 (—CH, d, J=4.5 Hz, 1H), 4.05 (—CH, dd, J=9.7, 3.1 Hz, 1H); MS (ESI) m/z: 172.1 [M+H].sup.+.

[0155] The synthetic method of the compound 29 in the present invention is similar to that of the compound 28.

[0156] The specific compounds synthesized and names thereof are as follows.

TABLE-US-00002 No. Structure Chemical name and analysis data 29 [00028] (S)-2-amino-1-((R)-2-methyloxirane-2-yl)-3- phenylpropan-1-one 2,2,2-trifluoroacetate Yield 76%, m.p.: 118-119° C.; .sup.1H NMR (400 MHz, CDCl.sub.3): δ 1.55 (—CH.sub.3, s, 3H), 2.99 (—CH, d, J = 4.4 Hz, 1H), 3.05 (—CH, dd, J = 14.3, 7.9 Hz, 1H), 3.24 (—CH, d, J = 4.4 Hz, 1H), 3.37 (—CH, dd, J = 14.3, 4.4 Hz, 1H), 4.29 (—CH, dd, J = 7.8, 4.6 Hz, 1H), 7.38-7.21 (—Ph, m, 5H); MS (ESI) m/z: 206.3 [M + H].sup.+.

III. Preparation of Compound of Formula (I)

[0157] The preparation of 2,5-di chlorobenzoyl-L-methyl serine- S-methyl-L-cysteinyl-methyloxirane (30) is taken as an example:

##STR00029##

[0158] The compound 7 (1 g, 2.4 mmol) and HOBt (0.5 g, 3.6 mmol) are dissolved in DCM. EDC.HCl (0.7 g, 3.6 mmol) is added. After stirring at −5° C. for 15 min, the compound 28 (0.7 g, 2.4 mmol) is added. DIPEA (1.2 mL, 7.2 mmol) is added after 15 min. Reaction is performed at low temperature for 25 min. After the reaction is complete at room temperature, the reaction solution is extracted with DCM. The organic phase is washed with 1N HCl, 5% NaHCO.sub.3 and saturated brine, and dried with anhydrous sodium sulfate. The solvent is evaporated to obtain the compound 30. The yield is 60%, m.p.: 60.8-62.3° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6 0.97-0.84 (m, 6H),1.35-1.23 (m, 2H), 1.52-1.47 (m, 3H), 1.70-1.58 (m, 1H), 2.18-2.06 (m, 3H), 2.84-2.72 (m, 1H), 2.87 (dt, J=10.4, 5.2 Hz, 1H), 3.09-2.93 (m, 1H), 3.31-3.25 (m, 1H), 3.49-3.38 (m, 3H), 3.71-3.55 (m, 1H), 4.04-3.87 (m, 1H), 4.69-4.50 (m, 2H), 4.85-4.71 (m, 1H), 7.18 (dd, J=11.0, 4.7 Hz, 1H), 7.76-7.63 (m, 1H), 7.40-7.32 (m, 2H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.94, 16.68, 21.17, 23.29, 25.15, 35.91, 39.87, 50.50, 52.08, 52.34, 53.57, 59.06, 59.32, 71.19, 129.04, 130.17, 131.45, 131.69, 133.31, 135.37, 165.24, 169.12, 169.95, 208.06; HRMS calcd for C.sub.24H.sub.33Cl.sub.2N.sub.3O.sub.6SNa, [M+Na].sup.+ 584.1359, found 584.1303.

[0159] The synthetic methods of all compounds in the present invention are similar to that of the compound 30.

[0160] The specific compounds synthesized and names thereof are as follows.

TABLE-US-00003 No. Structure Chemical name and analysis data 31 [00030] N-5-methylisoxazolyl-3-formyl-O-methyl-L-seryl-S- methyl-L-cysteinyl-L-leucyl-methyloxirane Yield 78%, m.p.: 70.2-72.7° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.95-0.90 (m, 6H), 1.34-1.22 (m, 2H), 1.50 (s, J = 5.4 Hz, 3H), 1.71-1.58 (m, 1H), 2.10 (s, 3H), 2.47 (s, J = 0.8 Hz, 3H), 2.79 (ddd, J = 14.0, 10.6, 6.6 Hz, 1H), 2.87 (t, J = 6.1 Hz, 1H), 3.01-2.90 (m, 1H), 3.42 (s, 3H), 3.69-3.53 (m, 1H), 3.96-3.80 (m, 1H), 4.65 (s, 2H), 4.76- 4.64 (m, 1H), 6.42 (s, 1H), 6.98 (s, J = 13.6, 6.6 Hz, 1H), 7.57 (s, 2H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.32, 15.87, 16.66, 21.23, 23.34, 25.13, 35.86, 39.92, 50.45, 52.04, 52.32, 52.76, 59.05, 59.29, 71.31, 101.28, 157.99, 159.42, 169.06, 169.99, 171.40, 208.05; HRMS calcd for C.sub.22H.sub.34N.sub.4O.sub.7SNa, [M + Na].sup.+ 521.2040, found 521.2226. 32 [00031] N-pyrazinyl-2-formyl-O-methyl-L-seryl-S-methyl-L- cysteinyl-L-leucyl-methyloxirane Yield 80%, m.p.: 50-51° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.99-0.89 (m, 6H), 1.42-1.17 (m, 2H), 1.51- 1.46 (m, 3H), 1.70-1.60 (m, 1H), 2.18-2.04 (m, 3H), 2.83-2.75 (m, 1H), 2.87 (dd, J = 6.5, 3.8 Hz, 1H), 3.03- 2.92 (m, 1H), 3.35-3.21 (m, 1H), 3.51-3.40 (m, 3H), 3.74-3.59 (m, 1H), 4.01-3.86 (m, 1H), 4.66-4.51 (m, 2H), 4.76 (ddt, J = 12.4, 8.0, 3.1 Hz, 1H), 7.25-6.83 (m, 2H), 8.63-8.48 (m, 2H), 8.78 (s, J = 4.4, 2.1 Hz, 1H), 9.37 (s, J = 1.4 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.88, 16.68, 21.19, 23.37, 25.17, 35.81, 39.87, 50.68, 51.91, 52.07, 52.77, 53.15, 59.31, 71.51, 142.79, 142.84, 144.32, 144.39, 163.41, 169.50, 170.01, 208.10; HRMS calcd for C.sub.22H.sub.33N.sub.5O.sub.7SNa, [M + Na].sup.+ 518.2043, found 518.2042. 33 [00032] N-2,5-dichlorophenyl-2-formyl-O-methyl-L-seryl-S- methyl-L-cysteinyl-L-phenylalanyl-methyloxirane Yield 78%, m.p.: 50.7-52.3° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) 1.31-1.20 (m, 3H), 2.10-1.92 (m, 3H), 2.78- 2.64 (m, 1H), 3.00-2.79 (m, 2H), δ 7.71-7.67 (m, 1H), 3.26-3.15 (m, 1H), 3.47-3.35 (m, 3H), 3.60 (dt, J = 9.3, 6.3 Hz, 1H), 3.98-3.90 (m, 1H), 4.62-4.49 (m, 1H), 7.41- 7.33 (m, 2H), 4.77-4.67 (m, 1H), 5.47-5.33 (m, 1H), 7.25-7.16 (m, 5H), 7.28 (dd, J = 6.8, 1.4 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.59, 16.58, 35.56, 37.03, 51.95, 52.47, 53.11, 53.68, 59.27, 59.33, 71.12, 127.09, 127.22, 129.10, 129.32, 129.38, 130.23, 130.41, 131.55,131.72, 136.91, 166.84, 169.57, 170.75, 210.53; HRMS calcd for C.sub.27H.sub.31Cl.sub.2N.sub.3O.sub.6SNa, [M + Na].sup.+ 618.1202, found 618.1202. 34 [00033] N-5-methylisoxazolyl-3-formyl-O-methyl-L-seryl-S- methyl-L-cysteinyl-L-phenylalanyl-methyloxirane Yield 75%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.27 (s, J = 11.7 Hz, 3H), 2.02 (s, 3H), 2.49 (s, J = 3.9 Hz, 3H), 2.83-2.62 (m, 2H), 2.96-2.83 (m, 2H), 3.22- 3.07 (m, 1H), 3.30 (t, J = 4.8 Hz, 1H), 3.41 (s, J = 7.6, 3.9 Hz, 3H), 3.61-3.52 (m, 1H), 3.95-3.79 (m, 1H), 4.51 (dd, J = 12.7, 6.8 Hz, 1H), 4.73-4.62 (m, 1H), 4.82 (dd, J = 13.1, 8.5 Hz, 1H), 6.43 (dd, J = 9.8, 4.8 Hz, 1H), 7.25- 6.91 (m, 5H), 7.34-7.27 (m, 2H)7.58 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.27, 15.15, 16.42, 36.70, 37.12, 51.64, 52.39, 53.08, 54.08, 58.98, 59.57, 71.34, 100.03, 125.09, 127.19, 128.59, 128.66, 129.35, 136.40, 150.40, 160.48, 169.20, 169.54, 170.17, 210.53; HRMS calcd for C44H57N5O7Na, [M + Na].sup.+ 790.4150, found 790.4139. 35 [00034] N-pyrazinyl-2-formyl-O-methyl-L-seryl-S-methyl-L- cysteinyl-L-phenylalanyl-methyloxirane Yield 50%, m.p.: 46.1-47.6° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) 1.25 (s, 3H), 2.10-1.92 (m, 3H), 2.78-2.64 (m, 1H), 3.05-2.80 (m, 2H), 3.32-3.13 (m, 1H), 3.50-3.42 (m, 3H), 3.68-3.58 (m, 1H), 3.99-3.88 (m, 1H), 4.62- 4.46 (m, 1H), 4.78-4.65 (m, 1H), 5.49-5.27 (m, 1H), 7.15-7.06 (m, 1H), 7.24-7.16 (m, 3H), 7.37-7.28 (m, 1H), 8.69-8.43 (m, 2H), 8.86-8.73 (m, 1H), δ 9.52- 9.29 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.55, 16.54, 36.92, 37.29, 51.22, 51.94, 53.10, 54.51, 59.28, 59.38, 71.49, 125.53, 127.16, 127.46, 128.64, 129.33, 136.92, 142.78, 144.34, 145.12, 147.52, 161.68, 169.55, 171.14, 210.53; HRMS calcd for C25H31N5O6SNa, [M + Na]+ 552.1887, found 552.1889. 36 [00035] N-2,5-dichlorophenyl-2-formyl-S-methyl-L-cysteinyl-O- methyl-L-seryl-L-leucyl-methyloxirane Yield 78%, m.p.: 59.8-60.3° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) 0.93-0.84 (m, 6H), 1.25 (t, J = 3.5 Hz, 3H), 1.37 (d, J = 4.7 Hz, 2H), 1.57-1.54 (m, 1H), 2.25-2.15 (m, 3H), 2.66-2.51 (m, 1H), 2.87 (dd, J = 14.1, 10.6 Hz, 1H), 2.98 (dd, J = 13.0, 3.9 Hz, 1H), 3.42 (d, J = 12.5 Hz, 3H), 3.40-3.32 (m, 1H), 3.60-3.55 (m, 1H), 3.92-3.85 (m, 1H), 4.54 (ddd, J = 12.2, 10.6, 4.3 Hz, 1H), 5.06 (dddd, J = 27.6, 19.7, 13.7, 10.3 Hz, 2H), 7.34 (ddd, J = 3.1, 2.4, 1.8 Hz, 3H), 7.47 (dd, J = 2.1, 0.6 Hz, 1H), 7.69 (s, J = 2.2, 1.1 Hz, 1H), δ 7.93 (s, J = 21.3, 2.5 Hz, 1H), 13C NMR (101 MHz, CDC13) δ, 15.52, 15.67, 22.70, 22.82, 29.68, 35.47, 36.92, 51.10, 51.90, 53.65, 53.87, 53.90, 59.38, 71.13, 127.08, 128.60, 129.39, 130.17, 131.49, 131.82, 165.54, 169.33, 170.10, 210.35; HRMS calcd for C.sub.24H.sub.33Cl.sub.2N.sub.3O.sub.6SNa, [M + Na] 584.1359, found 584.1303. 37 [00036] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl-O- methyl-L-seryl-L-leucyl-methyloxirane Yield 80%, m.p.: 49.5-51.8° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.83 (s, 6H), 1.25 (s, 3H), 1.32 (d, J = 7.4 Hz, 1H), 1.50 (d, J = 6.2 Hz, 2H), 2.22 (s, 3H), 2.48 (s, 3H), 2.93 (dd, J = 34.2, 11.1 Hz, 3H), 3.32 (dd, J = 17.3, 9.0 Hz, 2H), 3.42 (s, J = 9.7 Hz, 2H), 3.83 (s, 1H), 4.77-4.47 (m, 3H), 6.41 (s, 1H), 7.11-6.92 (m, 2H) 7.62 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.35, 15.59, 16.42, 22.62, 22.80, 29.67, 35.43, 36.99, 51.12, 51.86, 53.08, 53.66, 53.86, 59.35, 71.40, 101.34, 169.30, 169.52, 170.00, 170.10, 171.58, 210.82; HRMS calcd for C.sub.22H.sub.34N.sub.4O.sub.7SNa, [M + Na].sup.+ 521.2040, found 521.2037. 38 [00037] N-pyrazinyl-2-formyl-S-methyl-L-cysteinyl-O-methyl-L- seryl-L-leucyl-methyloxirane Yield 50%, m.p.: 59.6-61.2° C.; .sup.1H NMR (400 MHz, CDCl3) 0.96-0.93 (m, 6H), 1.26 (dd, J = 13.3, 4.0 Hz, 2H), 1.33 (d, J = 4.7 Hz, 3H), 1.64-1.56 (m, 3H), 2.23- 2.18 (m, 3H), 3.08-2.95 (m, 2H), 3.40 (d, J = 5.6 Hz, 3H), 3.49-3.47 (m, 1H), 3.86-3.80 (m, 1H), 4.67-4.51 (m, 2H), 5.15 (td, J = 9.3, 4.6 Hz, 1H), 7.16 (dd, J = 44.1, 7.9 Hz, 2H), 8.62-8.49 (m, 2H), 8.83-8.76 (m, 1H), δ 9.40 (dd, J = 8.0, 5.2 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.60, 15.73, 22.62, 22.80, 22.90, 29.68, 37.03, 51.23, 51.76, 53.00, 53.49, 53.59, 59.36, 71.48, 142.84, 143.69, 144.37, 147.85, 163.82, 169.67, 170.00, 210.93; HRMS calcd for C22H33N5O6SNa, [M + Na].sup.+ 518.2043, found 518.2041. 39 [00038] N-2,5-dichlorophenyl-2-formyl-S-methyl-L-cysteinyl-O- methyl-L-seryl-L-phenylalanyl-methyloxirane Yield 80%, m.p.: 51.2-52.7 °C.; .sup.1H NMR (400 MHz, CDCl.sub.3) 2.16 (d, J = 2.9 Hz, 3H), 2.51-2.43 (m, 3H), 2.93 (ddd, J = 23.3, 12.0, 7.8 Hz, 3H), 3.21-3.14 (m, 1H), 3.29-3.23 (m, 3H), 3.56-3.44 (m, 1H), 3.84 (dt, J = 17.3, 8.6 Hz, 1H), 4.61 (ddt, J = 40.1, 33.3, 14.0 Hz, 3H), 6.44 (dd, J = 18.4, 3.1 Hz, 1H), 7.22-7.12 (m, 4H), δ 7.37 (dd, J = 16.0, 5.2 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 14.09, 15.71, 35.58, 36.83, 51.09, 52.83, 53.40, 53.67, 59.04, 59.09, 71.19, 127.00, 127.04, 128.57, 129.40, 129.44, 129.46, 129.50, 131.46, 131.54, 131.82, 133.35, 136.03, 165.50, 169.66, 169.95, 210.59; HRMS calcd for C.sub.27H.sub.31Cl.sub.2N.sub.3O.sub.6SNa, [M + Na].sup.+ 618.1202, found 618.1202. 40 [00039] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl-O- methyl-L-seryl-L-phenylalanyl-methyloxirane Yield 80%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) 1.25 (s, 3H), 2.25-2.18 (m, 3H), 3.07-2.90 (m, 3H), 3.23- 3.14 (m, 1H), 3.27 (dt, J = 11.6, 3.9 Hz, 3H), 3.43-3.34 (m, 1H), 3.83 (dd, J = 8.3, 3.2 Hz, 1H), 4.82-4.45 (m, 3H), 7.25-7.18 (m, 3H), 7.44-7.27 (m, 2H), 8.59 (dd, J = 3.6, 2.2 Hz, 1H), 8.83-8.78 (m, 1H), δ 9.45-9.35 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) 812.02, 15.66, 15.90, 36.86, 52.47, 52.90, 53.20, 53.60, 59.06, 59.11, 71.23, 100.38, 127.02, 128.57, 128.62, 129.45, 129.53, 136.04, 150.26, 161.66, 169.70, 169.74, 170.03, 210.52; HRMS calcd for C.sub.25H.sub.32N.sub.4O.sub.7SNa, [M + Na].sup.+ 555.1883, found 555.1881. 41 [00040] N-pyrazinyl-2-formyl-S-methyl-L-cysteinyl-O-methyl-L- seryl-L-phenylalanyl-methyloxirane Yield 50%, m.p.: 49.7-51.2° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) 1.25 (s, 3H), 2.23-2.17 (m, 3H), 3.02-2.90 (m, 3H), 3.22-3.16 (m, 1H), 3.31-3.26 (m, 3H), 3.41 (ddd, J = 11.1, 7.3, 4.3 Hz, 1H), 3.77 (td, J = 9.0, 4.1 Hz, 2H), 4.80-4.46 (m, 3H), 7.16 (ddd, J = 23.3, 14.7, 6.8 Hz, 5H), 8 7.42-7.33 (m, 3H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 14.10, 15.83, 35.54, 36.86, 52.47, 52.77, 52.90, 53.20, 59.06, 59.11, 71.23, 127.02, 128.57, 128.59, 129.42, 129.45, 136.07, 142.85, 144.38, 147.84, 169.61, 169.74, 169.95, 210.35; HRMS calcd for C.sub.25H.sub.31N.sub.5O.sub.6SNa, [M + Na].sup.+ 552.1887, found 552.1888. 42 [00041] N-2,5-dichlorophenyl-2-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-leucyl-methyloxirane Yield 78%, m.p.: 163.1-164.5° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.91 (dd, 6H), 1.23 (s, J = 11.3 Hz, 3H), 1.60-1.52 (m, 1H), 1.75-1.64 (m, 2H), 2.14 (d, 3H), 2.22 (d, 3H), 2.91-2.74 (m, 2H), 3.14-2.92 (m, 3H), 3.35-3.27 (m, 1H), 4.66-4.52 (m, 2H), 4.83-4.68 (m, 1H), 6.99 (s, J = 18.3, 8.2 Hz, 1H), 7.24 (s, 1H), 7.28 (d, J = 7.5 Hz, 1H), 7.37-7.35 (m, 1H), 7.55-7.49 (m, 1H) 7.75-7.63 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.78, 15.85, 16.72, 21.17, 23.30, 25.22, 35.85, 35.95, 39.77, 50.79, 52.11, 52.39, 53.01, 59.13, 128.81, 130.23, 130.89, 131.51, 131.78, 133.35, 165.22, 169.50, 169.87, 207.98; HRMS calcd for C.sub.24H.sub.33Cl.sub.2N.sub.3O.sub.5S.sub.2Na, [M + Na].sup.+ 600.1130, found 600.1132. 43 [00042] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-leucyl-methyloxirane Yield 60%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.94 (dd, 6H), 1.50 (d, J = 4.1 Hz, 3H), 1.69-1.63 (m, 1H), 1.71 (d, J = 11.9 Hz, 2H), 2.13 (d, J = 12.0 Hz, 3H), 2.20 (d, 3H), 2.48 (s, J = 0.7 Hz, 3H), 2.80 (td, J = 14.2, 6.9 Hz, 1H), 3.02-2.86 (m, 4H), 3.30 (t, J = 5.5 Hz, 1H), 4.66-4.52 (m, 2H), 4.76-4.68 (m, 1H), 6.41 (dd, J = 4.4, 0.8 Hz, 1H), 6.97 (s, 1H), 7.15 (s, J = 7.4 Hz, 1H), 7.64 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.35, 15.81, 15.93, 16.72, 21.22, 23.37, 25.21, 35.90, 35.99, 39.81, 50.77, 52.06, 52.38, 52.88, 59.13, 101.29, 159.36, 169.40, 169.91, 171.51, 207.97; HRMS calcd for C.sub.22H.sub.34N.sub.4O.sub.6S.sub.2Na, [M + Na].sup.+ 537.1811, found 537.1808. 44 [00043] N-pyrazinyl-2-formyl-S-methyl-L-cysteinyl-S-methyl-L- cysteinyl-L-leucyl-methyloxirane Yield 50%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.93 (dd, 6H), 1.26 (s, 3H), 1.61-1.54 (m, 1H), 1.67- 1.61 (m, 2H), 2.06 (d, J = 15.0 Hz, 3H), 2.22 (d, J = 6.4 Hz, 3H), 2.93-2.72 (m, 2H), 3.13-2.93 (m, 3H), 3.32- 3.22 (m, 1H), 4.67-4.50 (m, 2H), 4.83-4.67 (m, 1H), 8.58 (s, 1H), 8.79 (s, J = 4.2, 2.2 Hz, 1H), 9.37 (d, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.81, 15.99, 16.70, 21.05, 21.21, 23.38, 35.91, 36.14, 39.81, 50.84, 52.10, 52.45, 60.39, 142.81, 142.88, 144.37, 147.75, 163.27, 169.50, 169.96, 208.00; HRMS calcd for C.sub.22H.sub.33N.sub.5O.sub.5S.sub.2Na, [M + Na].sup.+ 534.1815, found 534.1815. 45 [00044] N-2-methylthiazolyl-5-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-leucyl-methyloxirane Yield 78%, m.p.: 150-151° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.93 (d, J = 4.5 Hz, 3H), 0.95 (d, 3H), 1.50 (d, 3H), 1.60-1.54 (m, 1H), 1.73-1.68 (m, 2H), 2.12 (d, 3H), 2.20 (d, 3H), 2.74 (d, J = 2.3 Hz, 3H), 2.84-2.78 (m, 1H), 2.95 (dd, J = 9.6, 4.2 Hz, 2H), 3.05-2.93 (m, 2H), 3.28 (t, J = 5.8 Hz, 1H), 4.67-4.51 (m, 2H), 4.77-4.67 (m, J = 7.7, 6.8, 5.6 Hz, 1H), 7.06 (s, 1H), 7.52 (s, 1H), 7.71 (s, 1H), 8.06 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.81, 15.99, 16.70, 21.05, 21.21, 23.38, 35.91, 36.14, 39.81, 50.84, 52.10, 52.45, 60.39, 142.81, 142.88, 144.37, 147.75, 163.27, 169.50, 169.96, 208.00; HRMS calcd for C.sub.22H.sub.34N.sub.4O.sub.5S.sub.3Na, [M + Na].sup.+ 553.1583, found 553.1580. 46 [00045] N-2,5-dichlorophenyl-2-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-phenylalanyl-methyloxirane Yield 78%, m.p.: 150-151° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.50 (s, 3H), 1.96 (s, 3H), 2.21 (s, 3H), 2.81- 2.68 (m, J = 18.9, 14.0, 7.9 Hz, 2H), 2.89 (dd, J = 6.2, 1.0 Hz, 1H), 2.94-2.91 (m, 2H), 3.06 (dd, J = 13.9, 5.4 Hz, 1H), 3.16 (dd, J = 14.0, 4.5 Hz, 1H), 3.33 (d, J = 6.6 Hz, 1H), 4.52-4.46 (m, J = 7.0, 5.2 Hz, 1H), 4.75-4.68 (m, J = 13.4, 6.7 Hz, 1H), 4.84-4.78 (m, J = 11.5, 3.5 Hz, 1H), 7.15 (d, J = 8.8 Hz, 2H), 7.18 (dd, J = 4.3, 2.6 Hz, 2H), 7.21 (d, J = 3.4 Hz, 2H), 7.24-7.22 (m, 1H), 7.24 (s, 1H), 7.38 (d, J = 1.5 Hz, 2H), 7.68 (dd, J = 3.5, 2.1 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.49, 15.70, 16.53, 35.64, 35.73, 36.80, 51.88, 52.49, 52.87, 53.24, 59.33, 127.10, 128.56, 129.08, 129.26, 130.21, 131.53, 131.82, 133.36, 135.17, 135.83, 165.23, 169.43, 169.70, 207.12; HRMS calcd for C.sub.27H.sub.31Cl.sub.2N.sub.3O.sub.5S.sub.2Na, [M + Na].sup.+ 634.0974, found 634.0974. 47 [00046] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-phenylalanyl-methyloxirane Yield 80%, m.p.: 149-150° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.49 (dd, J = 8.2, 2.6 Hz, 3H), 1.98 (d, J = 4.9 Hz, 1H), 2.18 (d, 3H), 2.05 (d, J = 1.9 Hz, 2H), 2.48 (dd, J = 6.2, 0.8 Hz, 3H), 2.83-2.61 (m, 3H), 2.89-2.84 (m, 1H), 2.93-2.89 (m, 1H), 3.00-2.94 (m, 2H), 3.20-3.09 (m, J = 12.9, 7.9, 4.8 Hz, 1H), 3.35-3.29 (m, 1H), 4.50 (dtd, J = 12.4, 7.1, 5.4 Hz, 1H), 4.73-4.59 (m, 1H), 4.88- 4.74 (m, 1H), 6.40 (d, J = 13.2, 0.9 Hz, 1H), 7.15-7.09 (m, 1H), 7.20-7.14 (m, 2H), 7.25-7.20 (m, J = 7.5, 3.7, 2.1 Hz, 1H), 7.32-7.27 (m, 2H), 7.64 (d, J = 7.0 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.36, 15.56, 15.86, 16.52, 35.67, 35.83, 36.88, 51.72, 51.83, 52.18, 52.56, 53.24, 59.31, 101.30, 127.14, 128.58, 128.59, 129.28, 129.31, 135.79, 157.90, 159.36, 169.35, 169.71, 171.54, 207.06; HRMS calcd for C.sub.25H.sub.32N.sub.4O.sub.6S.sub.2Na, [M + Na].sup.+ 571.1655, found 571.1656. 48 [00047] N-pyrazinyl-2-formyl-S-methyl-L-cysteinyl-S-methyl-L- cysteinyl-L-phenylalanyl-methyloxirane Yield 78%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.24 (s, 3H), 1.99 (d, J = 4.3 Hz, 1H), 2.04 (t, 2H), 2.18 (s, 3H), 2.85-2.61 (m, 2H), 2.96-2.85 (m, J = 10.5, 6.7, 6.0, 4.3 Hz, 2H), 3.06-2.96 (m, 2H), 3.21-3.10 (m, 1H), 3.33-3.27 (m, J = 12.1, 4.8 Hz, 1H), 4.57-4.44 (m, J = 7.0, 5.1, 3.6 Hz, 1H), 4.84-4.63 (m, 2H), 7.14 (dd, J = 5.0, 2.2 Hz, 1H), 7.18-7.16 (m, 1H), 7.22-7.19 (m, 1H), 7.23 (dd, J = 6.8, 1.9 Hz, 1H), 7.32-7.27 (m, 1H), 8.62- 8.53 (m, 1H), 8.82-8.75 (m, 1H), 9.41-9.32 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.55, 15.91, 16.52, 35.74, 36.10, 36.88, 51.72, 53.23, 59.27, 59.31, 60.40, 127.11, 127.15, 128.54, 128.57, 129.29, 135.73, 142.83, 143.67, 144.33, 147.73, 163.23, 163.41, 169.72, 207.09; HRMS calcd for C.sub.25H.sub.31N.sub.5O.sub.5S.sub.2Na, [M + Na]+ 568.1658, found 568.1660. 49 [00048] N-2-methylthiazolyl-5-formyl-S-methyl-L-cysteinyl-S- methyl-L-cysteinyl-L-phenylalanyl-methyloxirane Yield 80%, m.p.: 73-74° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.49 (d, 3H), 1.96 (s, 2H), 2.03 (s, 1H), 2.20 (d, 3H), 2.73 (s, 1H), 2.74 (s, 3H), 2.87-2.77 (m, 2H), 2.94- 2.88 (m, 2H), 3.02-2.95 (m, 1H), 3.16 (dt, J = 13.9, 5.2 Hz, 1H), 3.31 (dd, J = 4.7, 2.3 Hz, 1H), 4.58-4.45 (m, 1H), 4.72-4.59 (m, 1H), 4.87-4.75 (m, J = 13.6, 8.5, 4.7 Hz, 1H), 7.15-7.01 (m, 1H), 7.20-7.15 (m, 3H), 7.25- 7.21 (m, 2H), 7.31-7.27 (m, 2H), 8.08 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.56, 15.68, 16.52, 19.70, 35.73, 36.04, 36.88, 51.98, 52.33, 52.49, 53.23, 59.31, 127.15, 127.17, 128.59, 129.28, 129.33, 132.94, 135.77, 143.73, 160.54, 169.68, 169.75, 170.95, 207.19; HRMS calcd for C.sub.25H.sub.32N.sub.4O.sub.5S.sub.3Na, [M + Na].sup.+ 587.1427, found 587.1428. 50 [00049] N-5-methylisoxazolyl-3-formyl-O-methyl-L-seryl-glycyl- L-leucyl-methyloxirane Yield 80%, m.p.: 47.7-49.1° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.91 (s, 6H), 1.23 (d, J = 10.5 Hz, 1H), 1.49 (s, 3H), 1.71-1.56 (m, 2H), 2.48 (s, J = 0.8 Hz, 3H), 2.88 (d, J = 4.9 Hz, 1H), 3.32 (d, J = 11.9, 5.0 Hz, 1H), 3.42 (s, J = 1.1 Hz, 3H), 3.67-3.55 (m, 1H), 3.97-3.73 (m, 2H), 4.21- 3.99 (m, 1H), 4.76-4.52 (m, 2H), 6.42 (s, J = 2.6, 0.9 Hz, 1H), 6.65 (s, J = 8.3 Hz, 1H), 7.12 (s, 1H), 7.65 (s, J = 6.0 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.34, 16.66, 21.19, 23.33, 25.04, 39.92, 43.02, 50.14, 52.42, 53.19, 59.05, 59.35, 71.48, 101.38, 158.00, 159.49, 168.54, 169.62, 171.49, 208.78; HRMS calcd for C.sub.20H.sub.30N.sub.4O.sub.7Na, [M + Na].sup.+ 461.2006, found 461.2005. 51 [00050] N-5-methylisoxazolyl-3-formyl-O-methyl-L-seryl-glycyl- L-phenylalanyl-methyloxirane Yield 85%, m.p.: 54-55° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.50 (s, 3H), 2.49 (s, 3H), 2.87-2.76 (m, J = 13.8, 8.1, 3.1 Hz, 1H), 2.90 (d, J = 4.9 Hz, 1H), 3.11 (dd, J = 13.9, 5.1 Hz, 1H), 3.30 (dd, J = 9.5, 4.9 Hz, 1H), 3.39 (s, 3H), 3.61-3.47 (m, 1H), 4.04-3.78 (m, 3H), 4.74- 4.60 (m, 1H), 4.91-4.76 (m, 1H), 6.41 (s, J = 0.8 Hz, 1H), 6.71 (d, J = 7.9 Hz, 1H), 7.09-7.01 (m, J = 13.2, 7.7 Hz, 1H), 7.20-7.10 (m, 2H), 7.25-7.20 (m, 1H), 7.29 (d, 1H), 7.63 (d, J = 7.0 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.34, 16.47, 37.08, 42.99, 52.48, 52.65, 52.95, 59.22, 59.27, 71.31, 101.38, 127.13, 128.56, 129.32, 135.58, 158.01, 159.48, 168.30, 169.65, 171.48, 207.55; HRMS calcd for C.sub.23H.sub.28N.sub.4O.sub.7Na, [M + Na].sup.+ 495.1850, found 495.1848. 52 [00051] N-5-methylisoxazolyl-3-formyl-glycyl-O-methyl-L-seryl- L-leucyl-methyloxirane Yield 80%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.92 (s, 6H), 1.50 (s, 3H), 1.59-1.52 (m, 1H), 1.72- 1.59 (m, J = 19.6, 15.6, 9.3, 4.0 Hz, 2H), 2.47 (s, 3H), 2.88 (d, J = 5.0 Hz, 1H), 3.27 (d, J = 5.0 Hz, 1H), 3.33 (s, 1H), 3.39 (s, J = 5.8 Hz, 3H), 3.80-3.72 (m, J = 9.1, 3.9 Hz, 1H), 4.14-4.09 (m, 2H), 4.62-4.53 (m, 2H), 6.42 (s, J = 0.8 Hz, 1H), 6.93 (d, J = 6.3 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 7.58 (s, J = 5.3 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.32, 16.73, 21.20, 23.31, 25.18, 39.82, 42.95, 50.77, 52.09, 52.42, 59.04, 59.13, 71.34, 101.36, 158.04, 159.74, 168.11, 169.90, 171.33, 208.20; HRMS calcd for C.sub.20H.sub.30N.sub.4O.sub.7Na, [M + Na]+ 461.2006, found 461.2005. 53 [00052] N-5-methylisoxazolyl-3-formyl-glycyl-O-methyl-L-seryl- L-phenylalanyl-methyloxirane Yield 80%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.48 (s, J = 11.6 Hz, 3H), 2.47 (s, 3H), 2.85-2.77 (m, 1H), 2.91 (dd, J = 4.9, 2.5 Hz, 1H), 3.17-3.10 (m, 1H), 3.23 (s, 1H), 3.29 (s, 3H), 3.34 (dd, J = 92, 7.5 Hz, 1H), 3.77-3.60 (m, J = 19.1, 9.2, 3.9 Hz, 1H), 4.13-4.03 (m, 2H), 4.56-4.39 (m, J = 18.2, 7.0, 3.9 Hz, 1H), 4.82-4.71 (m, 1H), 6.39 (s, J = 0.8 Hz, 1H), 6.82 (s, J = 6.9 Hz, 1H), 7.08 (s, J = 7.7 Hz, 1H), 7.18-7.12 (m, 2H), 7.25-7.20 (m, 1H), 7.31-7.27 (m, 2H), 7.52 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.32, 16.57, 36.79, 42.84, 51.92, 52.54, 53.17, 59.03, 59.32, 71.06, 101.37, 127.12, 128.55, 129.33, 135.61, 158.05, 159.68, 168.11, 169.68, 171.33, 207.11; HRMS calcd for C.sub.23H.sub.28N.sub.4O.sub.7Na, [M + Na].sup.+ 495.1850, found 495.1848. 54 [00053] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl- glycyl-L-leucyl-methyloxirane Yield 80%, m.p.: 58.4-59.8° C.; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.91 (dd, 6H), 1.26 (dt, J = 7.1, 2.6 Hz, 1H), 1.36-1.30 (m, 1H), 1.48 (s, 3H), 1.69-1.58 (m, 1H), 2.19 (s, 3H), 2.48 (s, J = 0.8 Hz, 3H), 2.88 (d, J = 5.8 Hz, 1H), 3.00 (qd, J = 13.9, 6.6 Hz, 2H), 3.31 (d, J = 4.9 Hz, 1H), 3.90 (dd, J = 16.9, 5.2 Hz, 1H), 4.11-4.01 (m, 1H), 4.74- 4.51 (m, 2H), 6.41 (dd, J = 4.4, 0.9 Hz, 1H), 6.71 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 5.5 Hz, 1H), 7.70 (d, J = 7.0 Hz, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 12.36, 15.89, 16.70, 21.16, 23.32, 25.09, 35.77, 39.76, 43.06, 50.38, 52.46, 52.54, 59.10, 101.32, 157.91, 159.49, 168.43, 169.97, 171.53, 208.73; HRMS calcd for C.sub.20H.sub.30N.sub.4O.sub.6SNa, [M + Na].sup.+ 477.1778, found 477.1781. 55 [00054] N-5-methylisoxazolyl-3-formyl-S-methyl-L-cysteinyl- glycyl-L-phenylalanyl-methyloxirane Yield 78%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.47 (s, J = 4.4 Hz, 3H), 2.17 (s, J = 3.2 Hz, 3H), 2.47 (s, J = 0.8 Hz, 3H), 2.85-2.77 (m, 1H), 2.91-2.88 (m, 1H), 3.02-2.92 (m, 2H), 3.12 (dd, J = 13.9, 5.0 Hz, 1H), 3.29 (d, J = 5.5 Hz, 1H), 3.82 (dd, 1H), 3.95 (dd, 1H), 4.68 (p, J = 6.8 Hz, 1H), 4.82 (td, J = 7.9, 5.0 Hz, 1H), 6.39 (s, J = 0.9 Hz, 1H), 6.76 (d, J = 7.6 Hz, 1H), 7.16- 7.11 (m, 3H), 7.25-7.21 (m, 1H), 7.28 (d, J = 1.6 Hz, 1H), 7.69 (d, J = 7.2 Hz, 1H); .sup.13C NMR (101 MHz, CDCl3) δ 12.35, 15.83, 16.49, 35.77, 36.95, 43.00, 52.32, 52.51, 52.82, 59.26, 101.32, 127.14, 128.57, 129.28, 135.61, 157.93, 159.44, 168.17, 169.98, 171.50, 207.58; HRMS calcd for C.sub.23H.sub.28N.sub.4O.sub.6SNa, [M + Na]+ 7511.1621, found 511.1619. 56 [00055] N-2-methylthiazolyl-5-formyl-S-methyl-L-cysteinyl- glycyl-L-leucyl-methyloxirane Yield 80%, oily liquid; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.91 (d, J = 5.7 Hz, 3H), 0.93 (d, J = 3.4 Hz, 3H), 1.51 (s, 3H), 1.57-1.51 (m, 1H), 1.71-1.60 (m, 2H), 2.18 (s, 3H), 2.73 (s, 3H), 2.88 (dd, J = 7.8, 5.1 Hz, 2H), 2.98- 2.94 (m, 1H), 3.36-3.24 (m, 1H), 3.89 (dd, 1H), 4.08 (dd, 1H), 4.73-4.56 (m, 2H), 6.82 (s, J = 8.0 Hz, 1H), 7.34 (s, J = 5.4 Hz, 1H), 8.10 (s, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 15.70, 16.72, 19.66, 21.16, 23.33, 25.14, 35.77, 39.70, 43.09, 50.50, 52.46, 52.63, 59.11, 132.98, 143.84, 160.81, 168.47, 170.56, 171.00, 208.93; HRMS calcd for C.sub.20H.sub.30N.sub.4O.sub.5S.sub.2Na, [M + Na].sup.+ 493.1549, found 493.1547. [00056]

PART II DETERMINATION OF PROTEASOME INHIBITION ACTIVITY

I. Proteasome Inhibition Activity

[0161] The present invention uses a fluorescent polypeptide substrate Suc-Leu-Leu-Val-Tyr-AMC (abbreviated as Suc-LLVY-AMC, Suc stands for succinyl group, AMC stands for 7-amide-4-methylcoumarin) to determine the chymotrypsin-like enzyme activity of the proteasome.

[0162] The proteasome used in the present invention is human red blood cell 20S proteasome, and the enzymes, fluorescent substrates and test buffers are all purchased from Enzo company. The experimental system is 16 μL, wherein the substrate is 8 μL, the proteasome is 4 μL (0.8 ng), the final concentration is 50 μM, the drug (inhibitor) is 4 μL, the final concentration is 2×10.sup.−6 M−4.88×10.sup.−10 M, the last concentration is 0 M, the actual preparation concentration is 8×10.sup.−6 M−1.95×10.sup.−9M, and the last concentration is 0 M. The specific experiment process is as follows:

[0163] 1. Drug Preparation:

[0164] The drug is weighed and added to DMSO to dissolve to a concentration of 10.sup.−2 M. 2 μL of the drug solution is pipetted and added to 98 μL of DMSO to obtain a drug solution with the concentration of 2×10.sup.−4 M. Then 8 μL of the drug solution with the concentration of 2×10.sup.−4 M is pipetted and added to 198 μL of H.sub.2O to obtain a drug solution with the concentration of 8×10.sup.−6 M. Drugs with the concentrations of 2×10.sup.−6 M, 5×10.sup.−7 M, 1.25×10.sup.−7 M, 3.12×10.sup.−8 M, 7.8×10.sup.−9 M, and 1.95×10.sup.−9 M are obtained by the same method, and the last concentration 0 M means drug-free.

[0165] 2. Substrate Preparation:

[0166] 25 mg of fluorescent polypeptide substrate is dissolved in 654 μL of DMSO to obtain a 50 mM stock solution. The stock solution is stored at −20° C., and diluted 500 times when in use. 8 μL of the diluted stock solution is added to each sample so that the final substrate concentration in the reaction system is 50 μM.

[0167] 3. Reaction System Preparation:

[0168] The 20S proteasome is diluted with a buffer solution to a concentration of 2 ng/μL from 8 ng/μL, and the diluted proteasome solution is added to a 384-well fluorescence microplate, 4 μL each. Then 4 μL of the sample to be tested is added to each well, the marketed drug Carfilzomib is used as the positive control drug, and reaction is performed at 37° C. for 15 min. After the reaction is complete, 8 μL of fluorescent substrate is added to each well, and the reaction is performed for 1 h at 37° C. in the dark. A 360 nm/460 nm fluorescence microplate reader (BMG LABTECH POLARstar OPTIMA Microplate Reader) is used to test the fluorescence value.

[0169] 4. Data Processing:

[0170] The fluorescence values of the products obtained under the action of the drugs of different concentrations after subtracting the substrate are calculated. The IC.sub.50 concentration of the drug to inhibit the proteasome is calculated by the GraphPad Prism software.

TABLE-US-00004 Compound No. IC.sub.50 (nM) 30 59.31 31 90.38 32 2797 33 31.66 34 91.85 35 1275 36 41.91 37 93.35 38 >2000 39 167.2 40 34.24 41 751.70 42 15.88 43 16.43 44 1484 45 71.49 46 50.82 47 27.32 48 >2000 49 35.53 50 69.22 51 57.04 52 1494 53 1319 54 27.15 55 23.56 56 123.6 Oprozomib 235.2

II. Cell Strain Inhibition Activity

[0171] The test solution used in the present invention is a single solution cell proliferation test kit from Promega company; and the cells used are U266 and RPMI8226. The experimental system is 110 μL, containing 90 μL of cell suspension, 10 μL of test solution, and 10 μL of drug (inhibitor). The final concentration is 4.54×10.sup.−8 M−1.77×10.sup.−9 M, the last concentration is 0 M, the actual preparation concentration is 5×10.sup.−7 M−1.95×10.sup.−8 M, and the last concentration is 0 M. The specific experiment process is as follows:

[0172] 1. Drug Preparation:

[0173] The drug is accurately weighed and added to DMSO to dissolve to a concentration of 10.sup.−2 M. 1 μL of the drug solution is pipetted and added to 199 μL of DMSO to obtain a drug solution with the concentration of 5×10.sup.−5 M. Then 3.3 μL of the drug solution with the concentration of 5×10.sup.−5 M is pipetted and added to 326.7 μL of serum-free RPMI1640 medium to obtain a drug solution with the concentration of 5×10.sup.−7 M. The drug solution receives gradient dilution by 1.5 times to obtain drugs with the concentrations of 3.3 ×10.sup.−7 M, 2.2×10.sup.−7 M, 1.48×10.sup.−7 M, 9.87×10.sup.−8 M, 6.58×10.sup.−8 M, 4.38×10.sup.−8 M, 2.92×10.sup.−8 M and 1.95×10.sup.−8 M, and the last concentration 0 M means drug-free.

[0174] 2. Cell Suspension Preparation:

[0175] After the cells are counted separately, dilution and preparation are performed so that the number of U266 is 1×10.sup.4 cells/well, and the number of RPMI8226 is 1×10.sup.4 cells/well.

[0176] 3. Reaction System Preparation:

[0177] 90 μL of cell suspension is added to each well of a 96-well fluorescence microplate, and incubated for 24 h. Then 10 μL of the sample to be tested is added to each well, the drug Oprozomib is used as the positive control drug, and incubation is performed for 24 h. After the reaction is complete, 10 μL of test solution is added to each well and incubation is performed for 2-3 h. The absorbance is tested with a 490 nm fluorescence microplate reader (BMG LABTECH POLARstar OPTIMA Microplate Reader).

[0178] 4. Data Processing:

[0179] The absorbances of the products obtained under the action of the drugs of different concentrations after subtracting the substrate are calculated. The IC.sub.50 concentration (nM) of the drugs to cytotoxicity is calculated by the GraphPad Prism software.

[0180] The results of some compounds are as follows:

TABLE-US-00005 No. U266B1 RPMI8226 30 25.19 NT 31 26.41 NT 32 13677 NT 33 31.66 51.87 34 28.05 29.6  35 422 NA 36 44.48 NT 37 51.52 NT 38 NA NT 39 54.21 82.28 40 58.48 NA 41 159.6 55.34 42 73.56 50.75 43 36.07 33.23 44 NA NA 45 55.91 56.25 46 59.31 58.51 47 44.16 32.78 48 NA NA 49 35.92 42.09 50 NT 70.24 51 NT 42.48 52 NT >100    53 NT NT 54 NT 66.36 55 NT 47.48 56 NT >100    Oprozomib 22.36 31.50 NT: Not tested; NA: Not active

III. Primary Cytotoxicity in Patients with Multiple Myeloma

[0181] In the present invention, blood cells from patients with multiple myeloma and blood cells from healthy volunteers are used to test the toxicity of candidate compounds. The test solution is a single solution cell proliferation test kit from Promega company. The cells used are CD138+ cells selected from the patients with multiple myeloma and monocytes from the blood of the healthy volunteers. The experimental system is 110 μL, containing 90 μL of cell suspension, 10 μL of test solution, and 10 μL of drug (inhibitor). The final concentration is 4.54×10.sup.−8 M−1.77×10.sup.−9 M, the last concentration is 0 M, the actual preparation concentration is 5×10.sup.−7 M−1.95×10.sup.−8 M, and the last concentration is 0 M. The specific experiment process is as follows:

[0182] 1. Drug Preparation:

[0183] The drug is accurately weighed and added to DMSO to dissolve to a concentration of 10.sup.−2 M. 1 μL of the drug solution is pipetted and added to 199 μL of DMSO to obtain a drug solution with the concentration of 5×10.sup.−5 M. Then 3.3 μL of the drug solution with the concentration of 5×10.sup.−5 M is pipetted and added to 326.7 μL of serum-free RPMI1640 medium to obtain a drug solution with the concentration of 5×10.sup.−7 M. The drug solution receives gradient dilution by 1.5 times to obtain drugs with the concentrations of 3.3×10.sup.−7 M, 2.2×10.sup.−7 M, 1.48×10.sup.−7 M, 9.87×10.sup.−8 M, 6.58×10.sup.−8 M, 4.38×10.sup.−8 M, 2.92×10.sup.−8 M and 1.95×10.sup.−8 M, and the last concentration 0 M means drug-free.

[0184] 2. Cell Suspension Preparation:

[0185] After the cells are counted separately, dilution and preparation are performed so that the number of CD138+ cells is 1×10.sup.4 cells/well, and the number of monocytes is 1×10.sup.4 cells/well.

[0186] 3. Reaction System Preparation:

[0187] 90 μL of cell suspension is added to each well of a 96-well fluorescence microplate, and incubated for 24 h. Then 10 μL of the sample to be tested is added to each well, the drug Oprozomib is used as the positive control drug, and incubation is performed for 24 h. After the reaction is complete, 10 μL of test solution is added to each well and incubation is performed for 2-3 h. The absorbance is tested with a 490 nm fluorescence microplate reader (BMG LABTECH POLARstar OPTIMA Microplate Reader).

[0188] 4. Data Processing:

[0189] The absorbances of the products obtained under the action of the drugs of different concentrations after subtracting the substrate are calculated. The IC.sub.50 concentration (nM) of the drugs to cytotoxicity is calculated by the GraphPad Prism software.

[0190] The results of some compounds are as follows:

TABLE-US-00006 Oprozomib 50 51 Patient 1 53.04 10.04 36.05 Patient 2 70.94 11.04 52.63 Healthy volunteer 3 598.23 >2000 >2000 Healthy volunteer 3 807.42 >2000 >2000 NA: Not active

IV. Pharmacodynamic (PD) Determination of Candidate Compounds

[0191] The preparation method of an intragastric administration solution is: A certain amount of the substance to be tested is precisely weighed and added into a glass bottle, and a certain volume of DMSO is precisely added to prepare a stock solution with a concentration of 20.0 mg.Math.mL.sup.−1. Then the stock solution is diluted with a polyethylene glycol 400 (PEG400) and citric acid buffer (pH=2.7) (1:1, v:v). The diluted solution is dissolved by ultrasound to make a dosing solution with the concentration of 1.00 mg.Math.mL.sup.−1. 24 ICR mice, weighing 18-20 g, are randomly divided into 4 groups. The 1.00 mg.Math.mL.sup.−1 candidate compound and Oprozomib are administered intragastrically to the mice as per 10.00 mg.Math.kg.sup.−1. Blood samples are collected before administration (0 min) and 2 min, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 12 h after administration, and about 0.200 mL of blood is collected from the orbit. The blood samples are placed in test tubes containing EDTA-K.sub.2, and centrifuged at high speed (7800×g) for 5 min, and then the plasma is separated and stored at −15° C. to −35° C. Analysis and test are performed in accordance with the instructions of the Proteasome-Glo test kit from Technical Bulletin company. Data is collected with BMG LABTECH software, and analysis and drawing are performed with GraphPad Prism 5.

[0192] Determination Method:

[0193] 1. 100 μL of whole blood is taken from each mouse. 500 μL of PBS is added to wash the cells. The plasma is removed by centrifuging (3000 rpm, 5 min, 4° C.), and the process is repeated twice. The solution is washed and then resuspended with 500 μL of PBS.

[0194] 2. The resuspended stock solution is diluted 5 times with PBS in two parallel gradients, and the diluted solution is added to a 384-well plate, 20 μL/well.

[0195] 3. After 20 μL of proteasome-Glo detection buffer (CHTL) is added to each well, the proteasome-Glo Kinet program in the microplate reader is operated, and testing is performed once every 1 min for a total of 60 min.

[0196] 4. 100 μL of the resuspended blood cells are taken. 100 μL of RIPA lysis buffer (containing 1% protease inhibitor) is added. The blood cells are lysed on ice (vortex oscillation is performed once every 5 min). Centrifugation is performed after 20 min (12500 rpm, 30 min, 4° C.).

[0197] 5. After centrifugation, the supernatant (4 μL) is taken and diluted 5 times (16 μL) with RIPA lysis buffer (without protease inhibitors) for total protein quantification. The BCA method is used, that is, 5 μL of the solution to be tested and 100 μL of the prepared BCA solution are added to each well of a 96-well plate, and two sets of parallel are made for each sample. After mixing by vortex, incubation is performed at 37° C. for 30 min. The BCAprotein program in the microplate reader is operated for direct test.

[0198] The pharmacodynamic results of the candidate compounds in ICR mice are shown in FIG. 1.

PART III EVALUATION OF THE EFFICACY OF CANDIDATE COMPOUNDS IN NUDE MICE

[0199] 1×10.sup.7 multiple myeloma RPMI-8226-leu cells are injected into the armpits of nude mice. After the tumor grows to an average volume of 50-100 mm.sup.3, the animals are randomly grouped according to the tumor volume and then administered. The 24 nude mice are divided into 7 groups: solvent control group (Control), positive control Oprozomib 50 mg/kg (1 time a day) group, compound 50 100 mg/kg b.i.d. (1 time a day) group, and compound 51 50 mg/kg (1 time a day) group, 5 mice in each group. Each group is given the test substances of the corresponding concentrations in the tail vein at a dosage of 10 mL/kg, and Oprozomib and compounds 50 and 51 are administered in the order of d1, d2, d8, d9, d15 and d16 for 21 days.

[0200] The tumor volume is weighed and measured 2 times a week. The relative tumor volume (RTV) and relative tumor proliferation rate (T/C) are calculated, and statistical analysis is performed by SPSS 19.0 software. The relative tumor volume (RTV), relative tumor proliferation rate (T/C) and tumor inhibition rate (IR) are calculated, and statistical testing is performed. The calculation formula is as follows:

[0201] (1) TV (tumor volume)=½×a×b.sup.2, where a and b respectively represent the length and width of the tumor;

[0202] (2) RTV (relative tumor volume)=V.sub.t/V.sub.0, where V.sub.0 is the tumor volume measured during group administration (i.e. d0), and V.sub.t is the tumor volume during each measurement;

[0203] (3) T/C (%)=T.sub.RTV/C.sub.RTV×100%, where T.sub.RTV is the RTV of the treatment group, and C.sub.RTV is the RTV of the solvent control group;

[0204] (4) IR (%)=100% T/C.

[0205] Preparation method of compound solution:

[0206] Preparation of 5% sulfobutyl-β-cyclodextrin:

[0207] 2.500 g of sulfobutyl-β-cyclodextrin powder is weighed and added into a beaker. 50 mL of citric acid buffer is pipetted into a beaker. The solution is transferred to a container after the powder is dissolved.

[0208] Preparation of compound 50:

[0209] 10 mg of compound 50 powder is weighed and added into a 4 mL centrifuge tube. 5% sulfobutyl-β-cyclodextrin is added to the compound 50 powder to 1 mL to obtain a 10 mg/mL compound 50 test substance solution. A certain amount of the 10 mg/mL compound 50 test substance solution is taken and diluted with 5% sulfobutyl-β-cyclodextrin to a 5 mg/mL test substance solution.

[0210] Preparation of compound 51:

[0211] 10 mg of compound 51 powder is weighed and added into a 4 mL centrifuge tube. 5% sulfobutyl-β-cyclodextrin is added to the compound 51 powder to 1 mL for complete dissolution by ultrasound to obtain a 10 mg/mL compound 51 test substance solution. A certain amount of the 10 mg/mL compound 51 test substance solution is taken and diluted with 5% sulfobutyl-β-cyclodextrin to a 5 mg/mL test substance solution.

[0212] Preparation of Oprozomib:

[0213] 10 mg of Oprozomib powder is weighed and added into a 4 mL centrifuge tube. 5% sulfobutyl-β-cyclodextrin is added to the Oprozomib powder to 1 mL to obtain a 10 mg/mL Oprozomib test substance solution. A certain amount of the 10 mg/mL Oprozomib test substance solution is taken and diluted with 5% sulfobutyl-β-cyclodextrin to a 5 mg/mL test substance solution.

[0214] The results of the in vivo efficacy of the candidate compounds in nude mice are shown in FIG. 2, and the results of the body weight change of the candidate compounds in nude mice after continuous administration are shown in FIG. 3.